Horticulture

Despite this more conserved estimate of genera, the floral resources used in New Hampshire are still rich compared to those utilized in Georgia. We also found that foraging females in Missouri foraged from more plants to form a single pollen provision mass than those in Georgia . This suggests that suitable floral resources at the time of brood provisioning may not be as diverse in Georgia as more northern areas of Ceratina’s range, or that they were simply not locally abundant in the area around the collected nests. Across its geographic range, C. calcarata encounters a broad variety of possible forage. Diets in Georgia, Missouri and New Hampshire were dominated by pollen from different plant genera . Out of the 96 floral genera found in provisions in this study, only Rubus was found in more than 1% of reads across all three states . All other genera, even if abundant in one or two states, procona buckets were rare in provisions from the third. For example, sumac was a key floral resource in New Hampshire but made up less than 10% of the reads in Georgia and was hardly utilized at all in Missouri .

It is important to note that while read counts have been correlated with microscopy pollen counts in many studies, factors such as pollen morphology can skew the abundance estimate obtained from DNA sequences. Our study uses the marker rbcl, which has shown strong correlation with pollen counts, outperforming trnL and ITS2. With this in mind, comparison of relative abundance between sites shows state-wise differences in diet. Many of these plant genera are common to all three states, so perhaps these dietary variations are due to differences in bee and floral phenologies, as well as possible microhabitat distinctions in floral assemblages in proximity to the bee nest. While we do not have data on floral distributions within each collecting site, our records of nest substrate allow us to determine that foraging was not skewed towards the host plants. Rubus was a common pollen source but even nests formed within Rubus plants did not show a bias in pollen collection. Different pollens vary in nutritional qualities, which may influence foraging decisions. Pollen can also have toxic constituents, and some generalist foragers appear to actively utilize a broad range of floral resources to alleviate the effects these may have on brood development. How these factors influence C. calcarata foraging is unknown but our results suggest that spatial orientation of floral resources alone does not determine foraging preferences.

The presence of a consistent core microbial community despite the variation in pollen sources suggests that many of the most common bacterial genera do not have specific floral associations. We identified a number of tentative bacteria–plant correlations, but these were not consistent among states . In the overall analysis, the tuplip tree genus Liriodendron was correlated with Lactobacillus, while the same plant genus was correlated with Sphingomonas in Georgia. In Missouri, Wolbachia was correlated with four plant genera: Brunia, Camptotheca, Rhus and Smilax but this bacterium was not correlated with plants in the other states or the overall analysis. The correlations found in Georgia and Missouri also differ to those previously identified in New Hampshire, following the same methodology. These correlations broadly suggest that plants and bacteria are co-occurring but the variance in results between the overall dataset and the state-level analyses indicates these relationships are facultative or transient. Using read data to identify co-occurrence correlations is statistically challenging and further experiments sampling pollen bacterial communities with and without pollinator visitation, such as the study by McFrederick et al., are needed to directly test for plant–bacteria associations. Whether plants harbor certain microbes over others or not, there are many factors altering microbial floral communities.

Long-term artificial warming of grassland plots was found to alter the microbial communities of plant leaves, including microbial groups common to bees. Aydogan et al. found Acinetobacter and Wolbachia increased in frequency, while Sphingomonas frequency decreased, these three bacterial genera being common to C. calcarata pollen provision and adult gut microbiomes. These temperature based microbial changes could translate into changes in insect microbiomes, and indeed climate has been correlated with changes in microbiome composition in some species such as the red palm weevil, the chestnut weevil and a spider mite. Flower visitation by bees can transfer microbes to flowers, but herbivorous insects, other pollinators including thrips and wind are thought to contribute to microbe dispersal as well. Similarly, the presence of potentially predatory or competitive species such as ants can reduce floral visitation and this in turn alters the microbes present on flowers. Any and all of these could be important factors influencing the observed microbiome variation in C. calcarata and are important considerations when concerned with wild bee health generally. Our study shows that the diet of C. calcarata varies widely with geography, with only Rubus found in more than 1% of reads at all three sites, indicating that this generalist bee species is able to utilize different resources as floral communities change. However, it seems that floral preference may not be simply determined by the proximity of the floral resource to the nest. The same six bacterial genera consistently dominated provisions in all sites but the relative abundance of these fluctuated widely. There are still many unknowns regarding how microbes are acquired, both in the pollen provisions and subsequently the bees themselves. Flowers appear to be general points of bacterial transmission,but so far specific associations have not been identified. The current lack of knowledge on microbial associates is a major hindrance in our ability to maintain diverse wild bee populations.When I took on the role of Preschool Program Director at Daybreak Star Preschool, an entity of United Indians of All Tribes Foundation , located in Daybreak Star Indian Cultural Center, I also took on the community that built and sustained a palpable culture of resiliency, passion, honesty, strength, and love. UIATF serves urban Natives and Alaskan Indians in the greater Seattle area, once a relocation city for American Indians. As an organization, we provide holistic human wraparound services beginning with a doula and prenatal care program all the way up to our Elders program. The preschool is right in the middle, not just programmatically, but at the heart of the organization. I entered at a time just after the onset of the COVID-19 pandemic in September 2020. The new school year had just begun, and I was coming from fourteen years as an early childhood educator, twelve of those years as a toddler and family educator, at the same school, made up of mostly affluent white families. The state of the world was unknown and uncomfortably new, so I thought, why not take on an entirely different kind of job with much more responsibility? Being in this new role as a director wasn’t the only newness of this journey; this would be the first time in a while that I would be surrounded by people who looked more like me than not. UIATF is made up of a racially diverse group of people who are also diverse in their upbringings, cultures, languages, and socioeconomic status; many of these people are also of Native American descent. The shroud of uncertainty and social trauma maintained its presence for some time, procona florida container as the various programs needed to figure out how they were going to maintain the delivery of service.

Questions about funding sources lingered as funders also had to figure out their own systemic approach to sustain the programs they supported. The tight-knit community of UIATF, specifically those working at Daybreak Star Indian Cultural Center, was and is the force that prevented everything from imploding on itself. At the onset of the lockdown, with about one to two weeks’ time, the remaining preschool staff made a quick pivot to open up care for essential workers, eventually opening up to other families with many of the heavy regulations we all experienced in one way or another. Likewise, other programs of UIATF found themselves with a unique opportunity to distribute funding to community members who needed rental assistance, gas vouchers, grocery vouchers, etc. The preschool program, like many other programs within UIATF, was greatly impacted by the pandemic and it became apparent that a great deal of healing and rebuilding would need to be done. How would this healing process begin? Who would facilitate this process? From whom do we have to learn lessons? The answer lies with our plant relatives. As with most answers to big daunting questions, the answer to our communal healing surrounded us, for plants carry wisdom if we’re ready to take it in. All around Daybreak Star Indian Cultural Center are probably thousands of plant varieties—some indigenous and some not, some invasive. The preschool has several growing boxes and a small plot where tobacco and strawberries grow alongside one another. And of course, blueberry season is highly anticipated among children. Beyond our little garden are communities of alder trees, groves of cedar towering above the trails that meander around Daybreak Star, and many conifers that share their cones and branches for various lessons and activities. In this article, I aim to illustrate the community inquiry we dove into while outlining lessons learned as they relate to the wisdom of our plant relatives. The focus and intention on the teaching from our plant relatives comes from Plant Teachings for Growing Social-Emotional Skills, a toolkit of a book and a set of cards aimed at guiding individuals in their journey to deepening their capacities of self-reflection to lead healthier and resilient lives. This toolkit came to fruition through a collaborative effort between GRuB , Northwest Indian Treatment Center, and the Seattle Indian Health Board to aid individuals going through recovery from addiction but is applicable to early childhood learning centers. Throughout this article, I will introduce a plant relative and their lessons as they relate to my journey in our Community-Based Inquiry.In the early spring of 2021, my curiosity about finding ways to push the school forward was poked by the leaders of the Indigenous Early Learning Collaborative , Tarajean and Ethan Yazzie-Mintz, Joelfre Grant, and others with the provocation to embark on a journey of inquiry within our community setting. As I was ingratiating myself into the UIATF community as a newbie, I searched for questions within our preschool community I would want to explore alongside those who have been with UIATF. This search process was primarily observational, seeing and listening for issues and concerns within our early learning community of families, educators, elders, and children. A silver lining of the COVID-19 pandemic was that there was a significant level of understanding that spending time outdoors was an effective way to prevent the spread of infection. The already established preschool classrooms had dedicated much of their time to being outside, transplanting learning experiences and outcomes to the temperate climate of Seattle, Washington. Yet there was an increasing need for families and their children to have access to early care and education, and at the same time, the adults in children’s lives wanted to make sure their children were safe from potentially contracting COVID-19. These needs paired with the potential of elevating Daybreak Star Preschool into a new phase of evolution sparked my curiosity to home in on a community inquiry. Daybreak Star Indian Cultural Center, where the preschool is housed, is set in an urban forest which affords many opportunities for land-based learning. My natural state of curiosity beckoned me to wonder: What would an outdoor preschool class look like within an urban Native context? And why hasn’t this happened yet? In the Pacific Northwest, where we are located, outdoor preschool settings are on the rise. The North American Association for Environmental Education reports that there has been a twenty-fivefold increase in nature-based preschools just in this past decade. Currently, there are 585 nature-based preschools in the United States alone1. Our state, Washington, has become the first and only state to license outdoor nature based early learning centers. The growing trend in nature-based learning environments indicates a progressive movement to create early learning experiences that not only offer a counternarrative to how preschool is taught and experienced but also expand the opportunity to foster a sense of love and respect for the natural world in young children.

Because the response of bee visitors to land use change depends on species specific requirements and these pollinators also have variable effects on plants, understanding the effect of land use change on pollination services requires knowledge not only of which pollinator groups shift to the human-altered landscapes, but also the rate of pollination that those groups have on the plant species in those landscapes. Future research will benefit from looking at a wider range of plants with a different range of target pollinators and that flower earlier in the year to better tease out these hypotheses. If the patterns of bee visitation and seed set that we observed are indeed consistent across other plant species, the novel plant communities created in these human-altered landscapes and the generalist bee species that are favored in such landscapes will lead to a reduction in overall pollination services. Under the Biological Species Concept, speciation is the evolution of barriers to gene flow between sister lineages . Comparative tests of the geography and evolutionary forces driving speciation suggest that reproductive isolation evolves primarily via divergent natural or sexual selection accompanied by some degree of spatial isolation between populations .

Within this framework, plastic plant pot the process of speciation can be conceptualized as comprising three stages: niche expansion through the colonization of a new selective environment, ecological divergence, and the evolution of reproductive isolating barriers between lineages. Within species, populations in unique selective environments reflect all stages along this continuum, from maladapted sink populations to strongly isolated and locally adapted ecotypes . Such populations may become extinct, homogenized through gene flow, or remain partially isolated ecotypes; intraspecific divergence only rarely results in the evolution of a new, persistent biological species. Understanding the ecological and evolutionary processes that determine progress toward adaptive divergence and speciation remains a major goal of evolutionary biology . In this dissertation, I tested specific hypotheses about the processes operating at each of these stages, using acombination of stochastic simulations and field and greenhouse experiments with life history ecotypes of common monkeyflower Mimulus guttatus DC .The initial evolution and persistence of small populations is critical for understanding the process of speciation, particularly in sessile organisms such as plants. Gene flow has historically been viewed as a powerful force maintaining genetic cohesion throughout a species range, necessitating large-scale geographic isolation for speciation to occur .

However, botanists have argued that gene flow is sufficiently limited in many sessile organisms to allow smaller populations to evolve more or less independently, particularly at the periphery of a species range or under strong divergent selection . Contemporary evidence suggests that the spatial scale of speciation is in fact related to the scale of gene flow , and that new plant species are often formed in small, ecologically divergent populations . Speciation in initially small populations is fundamentally different from vicariant speciation in widely-distributed races because of the complex interplay of selection, genetic drift, inbreeding, and gene flow . Small populations face unique Demographic and genetic processes may limit or promote adaptation to novel environmental conditions. Such processes are fundamental to determining when and why adaptation to novel environments may fail, as at species range limits , or when successful adaptation promotes the invasion of non-native species or the evolution of reproductive barriers . Properties that influence the evolvability of species have been central to theory regarding the maintenance of sexual reproduction and outcrossing , as well as recent interest in predicting species’ responses to global environmental change . Historically, models of adaptation to novel environments focused on the genetic consequences of selection, gene flow, and genetic drift as well as demographic processes, such as immigration . However, recent work suggests that phenotypic plasticity may commonly play an important role in allowing population persistence in novel environments . Unlike adaptive evolution, phenotypic plasticity can immediately increase local fitness following colonization of a new environment .

Phenotypic plasticity may be particularly likely in harsh or stressful environments due to developmental instability or the expression of crypticgenetic variation , and can shift a population to within the “realm of attraction” of a new fitness peak . Phenotypic plasticity can evolve as an adaptive response to the range of environments typically encountered by an organism over space and time, or can be a passive response to stress; in either case it can be adaptive or maladaptive in a novel selective environment . Levin suggested that plasticity in traits affecting the mating system may have particularly important consequences for niche evolution in plants because the mating system has direct impacts on both the demographic and genetic properties of a population. Specifically, Levin suggested that niche evolution in plants may be facilitated if colonization of a novel environment is associated with increased self-fertilization via plasticity in floral traits or self-incompatibility systems. Diverse floral traits affect self-fertilization rate in plants, including the spatial and temporal separation of stigma and anthers, the proportion of cleistogamous flowers, and the expression of self-incompatibility mechanisms . Plasticity in one or more of these traits resulting in increased self-fertilization has been widely documented in response to environmental stress, including herbivory , pollen limitation , drought , heat , salt , and shade . Conversely, several studies have found either no plasticity in mating system or increased outcrossing in response to environmental stress . Thus, the magnitude, underlying traits, and environmental drivers of mating system plasticity appear to vary greatly among taxa. From a genetic perspective, increased self-fertilization may promote adaptation by acting as a partial reproductive barrier to maladaptive gene flow or by temporarily increasing genetic variation in traits under selection. Although self fertilization is associated with reduced genetic variation over long timescales , rapid increases in the self-fertilization rate are predicted to increase genetic variation temporarily by generating positive correlations between additive allelic effects within loci among offspring . Such allelic correlations may increase the rate at which genetic variance recovers following a bottleneck and the response to selection in populations with mixed mating . Self-fertilization may further increase genetic variation in traits with more complex genetic architectures by converting epistatic or dominance variance to additive genetic variance . Partial self-fertilization may also result in more rapid fixation of new, beneficial mutations and essentially produces assortative mating for traits under selection . From a demographic perspective, self-fertilization provides reproductive assurance by ensuring at least some reproduction when mates are limiting . Mate limitation may be severe in novel environments because of small population sizes and/or isolation. Plants with abiotic pollination mechanisms often exhibit density-dependent pollen limitation . In animal-pollinated systems, scarce ancestral pollinators or ineffective novel pollinators may limit outcross pollen availability in new environments . The importance of reproductive assurance during colonization is supported by the observation that isolated or peripheral plant populations often exhibit increased self-fertilization . Correspondingly, plastic planter pot self-fertilization is associated with invasiveness in annual weeds and larger range sizes in Collinsia . Despite these potential benefits of mating system plasticity for the persistence and adaptation of colonizing populations, sudden increases in self-fertilization rate are commonly associated with reduced fitness due to the expression and fixation of deleterious alleles that accumulate in previously outcrossing populations . If the segregating genetic load of an outcrossing population is high, a plastic increase in self fertilization rate could decrease the efficiency of selection on quantitative traits and increase the probability of extinction following colonization . In addition, inbreeding depression is often environmentally-dependent, and may be most severe in stressful or novel environments . Alternatively, the process of colonization itself may result in the fixation or purging of deleterious alleles if the number of colonists is small . Population bottlenecks increase the frequency of rare deleterious alleles, and may result in reduced fitness regardless of self-fertilization rate.

Given the diverse potential effects of mating system plasticity on the demography and evolution of colonizing populations, its overall consequences for niche evolution remain unclear. We used individual-based simulations to examine the evolutionary and ecological contexts in which mating system plasticity may promote or inhibit niche evolution. We tested the effect of a constant plastic increase in prior self-fertilization rate on population persistence and local adaptation by examining the evolution of a quantitative trait under stabilizing selection and the probability of extinction in a novel environment. Specifically, we focus on a region of genetic and demographic parameter space in which sink populations are in a ‘race’ to become locally adapted prior to extinction. To distinguish reproductive assurance, increased genetic variation, and reproductive isolation as potential mechanisms, we tested the effects of pollen limitation, strength of selection, genetic architecture, and maladaptive gene flow on both extinction probability and local adaptation. We examined the evolution of inbreeding depression and genetic load in the colonizing population to test whether consideration of deleterious mutations alters the effects of mating system plasticity on niche evolution. Although this model includes several assumptions based on plant reproductive biology , it could also be applied to hermaphroditic animal systems with mate limitation and/or gamete dispersal .We used stochastic simulations that track individual genotypes to model the evolution and demography of a colonizing sink population connected by gene flow with a source population. Our model is similar to that used by Holt et al. in which one-way migration occurs from a locally adapted source population to an initially maladapted sink population. However, we consider a plastic increase in self fertilization rate following colonization of the sink habitat, the potential for pollen limitation, and the evolution of inbreeding depression and genetic load due to deleterious mutations.The source and sink populations are composed of diploid and hermaphroditic individuals with discrete generations. Initially, the source population contains K individuals and the sink habitat is empty. The source population evolves for 1000 generations to reach mutation selection equilibrium before C individuals are randomly selected without replacement to colonize the sink habitat. Both populations evolve for 1000 generations following colonization or until the sink population goes extinct. The order of life-history events within each generation is: fertilization, selection, density dependence, dispersal, reproduction, and death. Prior to reproduction, individuals undergo selection. This selective period encompasses mortality at all life-history stages between fertilization and reproduction, including seed development, germination, and seedling growth. The fitness of an individual is determined by two forms of selection: optimizing selection on a quantitative trait, and purifying selection on deleterious mutations. Optimizing selection can be either directional or stabilizing, depending on the similarity of the average phenotype to the optimum.Immediately following colonization of the sink habitat, populations either begin to evolve towards the optimum phenotype and increase rapidly in size, or decline towards extinction . Population size is strongly associated with the average genotypic value, and extinction occurs only in maladapted populations with sizes far below K . Thus, density dependence only occurs once sink populations have begun to adapt, and the value of K does not affect colonization success. Extinction mainly occurs within the first 10 generations following colonization, and no populations become extinct after 50 generations . Given that the key demographic and genetic dynamics determining colonization success operate within the first 50 generations, we focus our results on this period. However, we also examine longer-term outcomes after 500 or 1000 generations to test whether these patterns change over time.Mating system plasticity has a profound and immediate effect on the genetic variance in colonizing sink populations . In general, an increase in self fertilization rate temporarily increases genetic variation relative to obligate outcrossing, but this effect decreases through time. Mixed mating maintains higher genetic variation than obligate outcrossing for tens to hundreds of generations following colonization, whereas obligate self-fertilization results in an initial spike in genetic variation that declines rapidly to levels below obligate outcrossing populations . This general pattern was observed across a range of parameter values, though the magnitude and duration of mating system effects on genetic variance depend on the genetic architecture of the quantitative trait . The effects of mating system plasticity on genetic variance have consequences for niche evolution when adaptation is limited by low genetic variation or strong selection. Under these conditions, the sustained increase in genetic variance under mixed mating allows a greater response to selection following colonization.

We used simple linear regressions to determine how soil moisture affected total pollen deposition, total pigment deposition, pollen deposition per open male flower, and pigment deposition per open male flower on bee-pollinated stigmas. Pollen deposition per open male flower was right-skewed and thus inverse transformed to improve normality. Pigment deposition and pigment deposition per open male flower from high-irrigation plants were right-skewed and thus square-root transformed to improve normality. Pigment deposition and pigment deposition per open male flower from low-irrigation plants were right-skewed and thus log10 transformed to improve normality of the residuals. We used a multiple logistic regression to assess how soil moisture and pollination affected fruit set, and a general linear model to assess how soil moisture and pollination affected seed set. To assess the effect of bee visitation on fruit set, we used a multiple logistic regression containing two fixed factors: Apis mellifera visitation rate and Eucera visitation rate. For this analysis, plastic pots large we used visitation rates from surveys of bees visiting all plants in the plot .

To determine the effect of pollen and pigment deposition on seed set in bee and hand-pollinated plants, we used separate general linear models containing total pollen deposition or proportion pigment from high-irrigation plants as the fixed factor.The irrigation treatment affected Cucurbita pepo plants to some degree in terms of floral traits. Male flower production and pollen production linearly increased with increasing soil moisture , but no other measured floral trait exhibited a significant response to manipulated soil moisture levels . Video recordings confirmed that honey bees and squash bees were the most frequent visitors of squash flowers . Although there were no significant effects of plant soil moisture on the visitation rate of Apis and Eucera on male flowers , we did observe contrasting patterns of visitation on female flowers . Summed across Apis visits, both the time spent drinking nectar and contacting floral stigmas increased with plant soil moisture, as well . For Eucera, in contrast, visitation rate on female flowers was independent of soil moisture ; cumulative time spent drinking nectar and contacting floral stigmas followed suit . Pollen and pigment deposition on the stigmas of bee-pollinated plants increased with increasing plant soil moisture .

For pollen deposition we observed a linear increase with increasing plant soil moisture , but this relationship was no longer significant when we controlled for the number of open male flowers available to bees on the day that each stigma was collected . Pigment originating from male flowers on plants in the high-irrigation group was deposited in increasingly greater amounts on the stigmas of female flowers as the soil moisture experienced by recipient plants increased . This relationship remained significant after we controlled for the availability of male flowers from high-irrigation plants on the day that each measurement was made . For fluorescent pigment originating from male flowers on plants in the low-irrigation group, pigment transfer to stigmas was independent of the recipient plant’s soil moisture . Pollen and pigment deposition for hand-pollinated stigmas are listed in Appendix 2.2. Fruit set was independent of plant soil moisture and also did not differ as a function of pollination mode : bee-pollinated mean and hand-pollinated mean . For bee-pollinated plants, however, the likelihood of fruit set increased with increasing honey bee visitation but not with increasing squash bee visitation . Seed set increased with plant soil moisture and was higher for bee-pollinated plants than for hand-pollinated plants . For bee-pollinated plants, we further observed that seed set increased with the amount of pollen deposited on stigmas and with the proportion of pigment originating from plants in the high-irrigation group . In contrast, neither relationship was significant for hand-pollinated plants .We tested three predictions concerning how specialist and generalist pollinators interact with plants grown under a gradient of soil moisture conditions and how pollinator-specific behaviors affect plant reproduction.

We received partial support for our first prediction. The frequency of floral visits by honey bees increased with increasing plant soil moisture for female flowers but not for male flowers; visit frequency by squash bees was independent of plant soil moisture for both female and male flowers. Moreover, increased visitation by Apis in female flowers included increased duration of stigmatic contact with increasing plant soil moisture. Our second prediction was met; stigmatic deposition of pigment from plants in the high-irrigation group increased with increasing plant soil moisture. Although we were not able to directly track pollen deposition from plants in the high-irrigation group, pigment and pollen deposition were highly correlated. Support for our third prediction comes from the comparisons of bee-pollinated versus hand-pollinated plants. As expected, seed set increased with increasing plant soil moisture for both bee-pollinated and hand-pollinated plants. For bee-pollinated plants, seed set increased with increasing pollen deposition and with increasing pigment from plants in the high-irrigation group. Hand-pollinated plants, in contrast, received an even mix of pollen from plants in the low-irrigation and high-irrigation groups irrespective of the plant soil moisture of the recipient plant . These plants did not exhibit relationships between seed set and pollen deposition nor between seed set and the amount of pigment originating from plants in the high-irrigation group. These results taken together suggest that pollen from plants in the high-irrigation group was of higher quality than pollen from plants in the low-irrigation group .Given that the goals of this study were to investigate how the behavior of pollinators mediates the reproductive performance of plants that experience a gradient of soil-moisture environments, we did not attempt to identify the mechanisms responsible for why pollinators exhibit selectivity. Although an earlier study in this same system found that flower size and nectar production increased with increasing plant soil moisture, we found no evidence for similar responses in the current study and assume that inter-annual differences in temperature , the timing of planting, or differences in the experiment itself are responsible. One possible explanation for our finding of increased visitation by honey bees to female flowers with increasing plant soil moisture may have been caused by the emission or composition of floral VOCs attractive to Apis changing with increasing soil moisture . Irrespective of the mechanism of attraction, Apis often exhibit a preference for female flowers of C. pepo as well as those of C. foetidissima . Given that pollen production increased in male flowers with increasing soil moisture, increased visitation to female flowers with increasing soil moisture by honey bees is sufficient to explain higher deposition of pollen from plants in the high-irrigation group given that exhibited cumulative duration of stigmatic contact increased with visitation. We relied on fluorescent pigments to examine pollen movement among plants in this experiment. As a proxy for pollen deposition, black plastic nursery pots the use of pigments is established in the literature and in our study seems well justified given the close correlation between pollen and pigment deposition .

Nonetheless, this approach, by itself, does not permit direct identification of pollen from plants in low-irrigation and high-irrigation groups once it is deposited on the stigma. In fact, total stigmatic pollen deposition was independent of plant soil moisture after correcting for male flower availability. This result differed from the positive effect of increased soil moisture of the recipient plant on the deposition of pigment from plants in the high-irrigation group after correcting for available male flowers. However, total pollen deposition included pollen from plants in both the low-irrigation and high-irrigation groups. Therefore, the inclusion of pollen from all soil moisture levels in the measurement of total pollen deposition likely canceled out the effects from the high-irrigation pollen, as we found that pigment from plants in the low-irrigation group did not increase with increasing plant soil moisture of the recipient plant . As with our analysis of pollen deposition, analysis of fruit set also did not allow for an assessment of the contribution of pollen from different sources. It is noteworthy, however, that fruit set increased with increasing visitation by honey bees and was independent of visitation by squash bees. Other work in this system has revealed that Eucera outperform Apis with respect to per visit contributions to fruit set . Thus, the contributions of honey bees to fruit set in the present study provides additional evidence that Apis transport pollen from male flowers of plants in the high-irrigation group to the female flowers of plants in the high-irrigation group and in doing so enhance plant reproduction. Squash bees, in contrast, indiscriminately visited flowers with respect to plant soil moisture and likely transported pollen from plants across the soil moisture gradient. The findings of this study seem likely to apply to other plant-pollinator systems. Generalist bees often forage preferentially for high-quality floral resources, such as more viable pollen , pollen with a high protein content , nectar of increased volume and concentration , and nectar with increased amounts of amino acids . These preferences, in turn, can benefit plants through increased pollinator visitation and pollen deposition. Kasagi and Kudo , for example, found that preferences exhibited by bumble bees for increased nectar resources resulted in reduced pollen limitation in Phyllodoce. However, generalists need to not only be attracted to high-quality resources, but they also need to increase stigmatic deposition of high-quality pollen to augment plant reproduction. Mu et al. , for example, assessed that preferences for flower color by flies and honey bees coincided with increased pollen viability, which led to greater reproductive success in Gentiana leucomelaena. Compared to generalists, specialist pollinators are often considered more effective . Yet during stressful environmental conditions, such as droughts, generalists may preferentially visit high quality resources while specialists do not. Exhibiting distinct floral constancy amongst conspecifics of high-quality could thereby allow generalist pollinators to mediate successful reproduction in stressed plants. However, if all conspecifics lack resources of high enough quality for generalists, then these pollinators may disregard stressed plants altogether, leaving specialists as the sole floral visitors with the job of pollination. Reductions in the availability of high-quality floral resources may thus turn out to be detrimental for pollinators as well as their floral hosts. Plants species faced with drought may therefore increasingly trade-off allocation to separate, and perhaps competing, floral functions in order to attract a diversity of pollinators. Chapter 2, in part, is currently being prepared for submission for publication of the material. Gambel, Jess; Holway, David A. The dissertation author was the primary investigator and author of this material. The importance of day length for plant photosynthetic carbon gain has been widely recognized. Duration of the nocturnal period may also be important. During the nighttime, periods of minimal transpiration allow water potential gradients among plant parts to dissipate by internally redistributing water to tissues of lower water potential, including the movement of water to roots in dry soil layers . Nocturnal internal hydraulic redistribution may be very important to the maintenance of roots in surface soil layers where root densities are highest, as surface soil can be dry for extended periods in many regions . However, the rate at which water redistributes at night depends on a combination of environmental and plant factors, including the magnitude of differences in water potential among tissues and the hydraulic conductivity of the vascular system . Many factors influence the rates of tissue rehydration of roots at night. Nocturnal transpiration due to incomplete closure of stomata or high vapour pressure deficits can create and maintain high water potential gradients between the soil and leaves and thus limit water movement to other parts of the root system. In addition,rehydration of roots in dry soil layers may be slower where there are significant constraints to water movement, including embolisms, small-diameter and curved xylem vessels with extensive branching , high frequency of pits and end plate membranes , and the presence of heartwood, latewood, and rays . Lateral water movement may also be limited by the high resistances that form at the stem base–root junction . Vaccinium corymbosum has been reported to be a species that does not effectively distribute water laterally . Using a split-root water application, Abbott and Gough found that dyes did not move laterally from one stem to another and observed root mortality in the unwatered root container. The main objective of this study was to quantify patterns of nocturnal internal hydraulic redistribution and conductances among the first seven orders of V. corymbosum roots under severe drought conditions and its implications for root tissue rehydration.

Such practices might include the use of noncrop vegetation, such as insectary plantings, which might provide natural enemies with a secondary aphid species as a food source before cabbage aphids are established in a broccoli field. Alternately, the pollen and nectar resources provided by such micro-habitats might also attract beneficial insects before a crop has become colonized by aphids. Nieto also notes that it would be advantageous to integrate spray policies in conventionally managed systems that recognize aphid location on the plant as an important contributor to broccoli harvest, rather than utilizing a presence-absence or zero-tolerance threshold once heading has begun. Nieto’s advisors for the research include Jeffrey Honda and Shannon Bros of San Jose State’s Department of Biological Sciences, Bill Settle of the United Nation’s Food and Agriculture Organization, Center director Carol Shennan, and Rachel O’Malley of San Jose State’s Environmental Studies Department.Blueberries are traditionally grown in colder climates, growing raspberries in container where the plants become dormant in winter.

Finding varieties that perform well under Central Coast conditions is one of the trial’s main goals. So far, it’s brought some surprises. “The varieties with the highest chill requirements [northern highbush varieties requiring a relatively long period of cold temperatures to produce fruit] are dormant this winter, while those with low chill requirements [southern highbush varieties] never entered dormancy and are wanting to make fruit year round,” said Leap. “I was expecting just the opposite to happen.” In order to create the acidic conditions that blueberries prefer, Leap added the per-acre equivalent of two tons of sulfur along with redwood mulch to the trial site’s soil prior to planting last January. A mix of peat moss and raspberry pomace was placed in the planting holes for additional acidity, and the plants were mulched with more redwood chips to a depth of over three inches. These efforts increased the soil’s acidity from its original pH of 5.9 to its current level of 5.1. In addition, organic matter levels rose from 2.5 to 3.5, and the CEC [a measure of nutrient availability] increased from 8 to 11. “We’ve created a nice environment,” said Leap, who also noted that “it’s a lot of work.” Still, the rewards can be high—a successful blueberry crop can generate $30,000 to $50,000 per acre, making it a potentially lucrative cash crop for small-scale organic producers.The centipede-like garden symphylan presents a vexing challenge for organic and conventional growers alike.

Measuring less than a 1/4-long, these mobile soil dwellers feed on the roots of developing plants, weakening or killing stands of both seedlings and developed crops, and potentially increasing their susceptibility to soilborne diseases. Conventional growers currently rely on organophosphate insecticides to control GS populations. However, the EPA is considering removing OPs from use due to their impacts on human health and the environment. Because garden symphylans also feed on decomposing organic material, they often flourish in well-managed soils with high levels of organic matter. Although there is no approved material for organic control, anecdotal evidence has demonstrated that symphylan populations are suppressed in areas where potatoes are grown. This observation holds promise for using potatoes in crop rotations to limit symphylan damage, and in isolating the potato compound that affects symphylans. Based on this observation, a field study is currently underway to test the suppressive effects of potatoes in controlled, replicated trials at the Center’s organically managed UC Santa Cruz farm and at four other sites in California and Oregon. The work is part of a study coordinated by Jon Umble of Oregon State University and funded by a grant from the USDA’s Western Sustainable Agriculture Research and Education Program. Umble and UCSC Farm manager Jim Leap established three replicates of three treatments: potatoes only, potatoes intercropped with corn , and corn only. Similar plots were established at the four other study sites in order to examine potato’s effect on GS in different soil and climate conditions. As predicted, GS population levels correlated directly with the cropping system: they were highest in the corn crop, lowest in the potato crop, and intermediate in the corn/potato intercrop.

According to Umble, these results provide strong evidence that GS populations do decrease in potatoes, and that this effect is fairly consistent over variable conditions. This spring, Leap will incorporate the winter cover crop planted on the study site and then directly sow broccoli, known to be an excellent host for symphylans. According to Leap, the broccoli will serve as an “indicator crop”—weakened or dying plants will indicate plots where GS populations are highest, while healthy stands will likely correlate with low GS numbers. The researchers will also sample for symphylans to confirm these indications. Ongoing work on the trial includes lab tests to determine which compounds in potatoes affect symphylans, and when the observed decrease in GS numbers occurs during an 8- week study period. Umble hopes that results from these studies will give insights into the mechanisms of the effects being observed in the field, and eventually offer growers an option for preventing GS damage.The Center has received a grant of $6,985 from the Stanley Smith Horticultural Trust to fund construction of new mist house facilities for asexual plant propagation at the Center’s Farm and Alan Chadwick Garden sites. The new mist houses will include polycarbonate growing tables, biotherm heating systems, hot water heaters, and mist/benchtop watering systems with auto controllers. These facilities will be used to teach apprentices and undergraduates the skills involved in propagating plants from cuttings. One challenge of asexual propagation using organic techniques is how to stimulate root growth without the synthetic rooting hormones used in conventional operations, and how to prevent mildew and other fungal growth without using synthetic fungicides. The Stanley Smith Horticultural Trust grant will fund a trial of non-synthetic alternatives to both rooting hormone compounds and fungicides for mist house use. Synthetic rooting hormones are not allowed under the USDA National Organic Program for the production of organic nursery stock. Garden managers Orin Martin and Christof Bernau plan to research available alternatives such as willow extract, kelp, and others. With assistance from the Center’s research staff, they will set up and monitor trials of several of these alternatives. Martin and Bernau also plan to trial a non-synthetic alternative to fungicides for the mist house, a commercially-available biological product called Root Shield. Grant writer Ann Lindsey also made a successful appeal to the Gaia Fund, which is providing $5,000 toward core support of the Center’s organic farming and gardening Apprenticeship training program in 2005. The Apprenticeship also received a $25,000 grant from an anonymous foundation to provide core salary support for the program in 2005.They should recognize safety and nutrition as consumers’ top concerns, but they should also devote attention to ethical issues, particularly the humane treatment of animals, raspberry container size environmental impacts, and social justice issues. Because respondents identified labels as their preferred source of information, eco-labels may be an appropriate way to address these matters. A majority of respondents indicated a willingness to pay more for strawberries that embodied a living wage and safe working conditions, even at price premiums up to 71% higher. The rapid growth of organic food sales, as well as sales of fair trade products from other countries, suggests that promoting the ethical values represented in food will continue to be a promising marketing strategy. Consumers who are interested in ethical aspects of the food system should recognize that their purchasing decisions can influence the way their food is grown, processed, and distributed. Unfortunately, there are currently few avenues for individual consumers to let producers and retailers know about their concerns. “You can talk to a store manager and hope to influence their decisions about what to stock or not stock. You can try to write to the company, but for the 10 corporations that sell over 50% of the food and drink in this country, they’re probably not going to be very responsive to one person,” said Howard.

Groups such as the Organic Consumers Association offer a way for individuals to make their voices heard through such mechanisms as letter-writing campaigns and other lobbying efforts on a variety of topics. “For example, OCA was a partner in the coalition that successfully campaigned to encourage Trader Joe’s to eliminate genetically engineered ingredients from their store-brand products,” said Howard. Shoppers can also seek out and support existing ecolabels, such as “Organic,” “Humane Husbandry,” “Free Farmed,” “Buy Fresh, Buy Local,” or the “Black Eagle” label, which identifies produce from farms that have contracts with the United Farm Workers Union, indicating “decent wages, benefits, and working conditions.” For consumers seeking information on eco-labels, Consumer Reports has an excellent web site that evaluates the wide variety of labels now being used on both fresh and processed food, body products, and household products. The web site currently includes information on 137 different labels and general claims, such as “cruelty free.” Howard believes that the fact that people want to know more about their food, yet individually do not have much voice in the food system, can be seen as an opportunity. Farms, cooperatives, and businesses involved in the food system can develop a way to give consumers input. For example, a subscription organic food delivery service in Denmark serves nearly 50,000 households. This “humongous CSA” employs people in a “conversations department” to handle more than 6,500 phone calls and 10,000 e-mails a month. This has helped them to grow at a rate of 10,000 subscribers a year while rapidly adjusting to what consumers want. By being more responsive to their needs, farmers, cooperatives, and retailers have the potential to provide consumers with the information they’re seeking and meet demands that are not being met by multinational corporations.Peaches and nectarines hail from northwestern China . The specific name persica is a misnomer, probably attributed to its spread via trade caravans from China into Iraq and Iran and eventually to Europe. The fruit came to the Americas with the Spanish explorers in the 16th century on their conquering expeditions. It was then spread across the U.S. by Native Americans. The nectarine is as old as the peach, with records of cultivation dating back to 2,000 BC. It is either a chance seedling or a whole tree mutation . Commercially, peaches and nectarines are grown at latitudes between 25º–45º North and South of the equator. Major peach growing regions include Chile, China, Northern Italy, Spain, Turkey, California, Southeastern U.S., New York, Pennsylvania, and Michigan. They can be grown closer to the equator than any other species of temperate zone deciduous fruits because of their tolerance for heat and humidity, and their low chill requirements for breaking dormancy. The peach, often referred to in old pomology texts as the “Queen of Fruits,” ranks only behind the apple in worldwide production and economic worth. Their sweet flavor, aroma, and nectar set the bar very high for sun-warmed tree-ripe perfection that evokes the essence of summer. Peaches are the shortest-lived of all deciduous fruit trees, with an average life expectancy of only 20–40 years . Because the genetics of the peach are much less variable than any other fruit, the trees of almost every seedling bear edible fruit. There are also more cultivars of peaches than any other fruit owing to the ease of obtaining quality seedlings from peach crosses. Peaches and nectarines can be grouped into two basic flesh types—clingstone and freestone. Clingstones exhibit a firm-textured flesh that cannot be pulled off the stone and must be cut away with a knife. Because they hold their shape when cut or sliced, they are the logical candidates for canning, drying, or being used fresh, halved, or sliced. Freestones are softer-fleshed varieties with higher juice content, and separate easily from the pit. They lend themselves to fresh eating. Additionally, peach tastes can be linked to flesh color and “old school” vs. “new school” varieties. Old school varieties don’t color evenly or have as bright a sheen to their skin. They have a more balanced sugar/acid ratio contributing to a fuller old-timey peach flavor. They have a very limited shelf life, must be tree ripened to have full flavor, and bruise easily, giving rise to that old farmers’ market adage, “Real peaches don’t stack.” These “old school” varieties include Suncrest, Elberta, Babcock, J.H. Hale, Red Haven, Le Grand, Rio Oso, Sun Grand, and Baby Crawford .

If more and more meat eaters are entering the market as it matures, then PBM is reaching those who are more likely to replace meat in their diet. Seeing evidence of PBM going mainstream the meat eaters would be encouraging for future growth. In order to test how the market of first time buyers is changing in a comparable way to buy and rebuy, I created a 1/0 variable of early and late. The cut off is set at April 29th, 2019 and while this may seem partly arbitrary, it closely matches when the public became much more aware of PBM. This was close to when Burger King first began running national ads for its Impossible Whopper, which attracted an unprecidented amount of attention for PBM. In addition, by this time, PBM had been available in all stores across this chain of grocery stores for at least 6 months, but this week marked the begining of a nationwide increase in demand for PBM in this grocery chain. We can see both of these trends in figure 1.1. On the left is a Google Trends search of Plant Based Meat for 2017 to 2019, raspberry container growing showing the large spikes in interest in the spring of 2019. On the left, I show the quantity of Plant Based Meat sold per store per week.

At the beginning, sales per store are high, but that may reflect the limited geographies that the item was made available in. As more and more stores started selling PBM, sales per store leveled out at a much lower level, until the spring of 2019, when sales began to rise again as nationwide interest in PBM picked up. In the next section, I will use week of first purchase as an alternate that can pick up more variation. Table 1.1 displays all the covariables discussed and how they relate to the three variables of interest, as well as t stats and p values. With such large sample sizes, the covariates on the variables of interest are unsurprisingly almost always statistically different between the two groups of households. Of particular interest for the buy / no-buy groups, those who bought PBM were more likely to be younger, to have a higher income, and to be shopping in a more Clinton heavy area. The extra variables created from the pre-period market basket are also interesting, if not completely surprising. While households were more likely to be “low meat” households that spent less than 5% of their pre-period budget on meat, the difference was only a half percentage point. This supports the often repeated fact that PBM customers also buy meat. However, they still bought less meat, with a larger gap in their average market basket raw percent devoted to meat in the pre period. In addition, buyers of PBM bought about 3 times as much veggie burgers and tofu.

So while PBM buyers did, on average, also buy meat, they bought less of it, and also bought a lot more traditional meat replacements. Because the number of buyers of PBM is oversampled, I cannot say what percent of all customers have bought it at least once. The early/late comparison and rebuyers in the second and third section of table 1.1 only include those who bought PBM once, so the difference in these groups would not necessarily be the same as between the buyers and no buyers. The demographics of rebuyers were only slightly different: skewing slightly younger and shopping in slightly more Clinton voting areas. However, rebuyers were more likely to be “low meat” purchasers and had spent more on traditional replacements and a little less on meat in the pre-period. That seems to infer that PBM was more likely to catch on with customers who were already interested in replacing meat in their diets. Finally, it’s important to note that less than a third of customers rebought PBM in the three months after first trying it. Because I have trimmed the sample to only include households that make regular trips , each customer had plenty of opportunities to buy PBM again, and this shows that PBM has not yet converted many of the people who try it to become regular purchasers. There are also some clear patterns in how earlier first time buyers compared to later ones. Early buyers were younger and less wealthy . Earlier buyers were more likely to be ’low meat’, bought less meat and ground beef, and bought more meat replacements. This suggest that later purchasers were less predisposed to buying tofu or veggie burgers, so were better candidates to be replacing thier meat intake, but comparing late buyers to those who never bought PBM is more mixed.

Later buyers are slightly less likely to have bought low amounts of meat than non PBM customers, but on average, they still spent less total on meat and ground beef and spent more on tofu and veggie burgers. This means that the most meat heavy consumers are still holding out on trying PBM.In order to test whether there was any evidence of plant-based meat competing with meat, I identified a random source of price variation in the data: region level price promotions . In private conversations with the grocery chain, they explained that promotions are almost exclusively decided on by each of 11 regional pricing executives, with very little input from the corporate office. For small, less important UPCs, such as the PBM UPCs, these individual pricing executives are unlikely to consult any pricing models or other data, but instead often simply put certain items on sale by intuition, or for the reason that those items had not been on promotion in a long time. In order to test this lack of coordination, I calculated the correlation of PBM products by region with other PBM products and all other meat products across all regions. While some larger UPCs had reasonable correlations across regions, nearly all PBM UPC correlations with themselves AND with other UPCs were unremarkable. In addition, I ran regressions of lagged quantity of PBM on price across each region individually, to see if some regions were putting PBM on promotion when either demand had been high or low in the weeks earlier, and found no significant results in any region. As a result, I felt confident in the assessment of my corporate contact at this chain that promotional prices for PBM are as good as random. This is especially true when looking at each specific store: decisions to promote are not related to store specific facts, as they are set at a higher regional level. Promotional prices are identified in the data by two variables: net price and gross price. Gross price is the price listed on the price tag in the store as the price when the item is not on promotion . If the item is on promotion, the gross price is the price listed as “was”, while net price is the price actually paid. In theory, all these prices are identical within each region each week, but sometimes there is unexplained variation in the prices that customers actually pay. This may be explained by manufacturer’s coupons, blueberry plant pot but other times it is unexplained. In order to get a region level weekly promotion, I subtract the gross price from the net price across all purchases within a region in each week.According with the International Diabetes Federation , the prevalence of people with T2DM raised from 108 million to 537 million in the last 40 years. T2DM is a disease caused by the incapacity of the cells to respond to insulin effects. This disease is strongly linked with obesity resulting from the excess of body weight and sedentarism . Obesity is considered a complex chronic progressive disease defined as abnormal or excessive adipose tissue accumulation caused by an imbalance of energy intake and energy expenditure that leads to mild, chronic, systemic inflammation. In a cross-sectional study from 2015 in Canadian adults, it was reported that ultra-processed food consumption, which contributed to 24-73% of total daily energy intake, was associated with higher prevalence of obesity in the studied population . It was also reported that high fat diets and elevated intake of red meat as well as high sugar intake impairs health causing obesity and increase the risk of noncommunicable diseases such as type 2 diabetes mellitus , dementia, myocardial infarction, stroke, hypertension, fatty liver disease and cancer . In general, obesity is associated with a lower life quality and expectancy by an estimated of 5-20 years depending on the severity and NCD’s . Adipose tissue has a crucial role on systemic energy balance , playing an essential role in maintaining lipid and glucose homeostasis.

The fat stored as triglycerides tends to accumulate in the subcutaneous and visceral depots, incrementing their size and producing hypertrophy, hyperplasia, and systemic metabolic dysfunction . The main mechanism believed to link obesity with T2DM is insulin resistance derived from adipose tissue accompanied by impaired insulin secretion by β-cells in the pancreas. Free fatty acids stimulate NF-κB and P38 MAPK signaling pathway through MyD88 and TRIF mediated downstream pathways with a subsequent activation of TLR4 expression in adipocytes and macrophages, increasing ER stress and producing ROS, promoting the secretion of proinflammatory cytokines, causing the initial step of low-grade systemic inflammation . The pro-inflammatory adipokines secreted by adipose tissue are monocyte chemotactic protein- 1 , tumor necrosis factor α , interleukin 1-β , and interleukin-6 . The potential cellular mechanisms of obesity-induced insulin resistance start with increased systemic TNF-α which stimulates the activity of IKK, p38 MAPK, JNK, and PKC, insulin receptor substrate ; impairing tyrosine phosphorylation and increasing the risk of insulin resistance in adipose tissues, muscles, and liver . Lifestyle modifications with controlled calorie intake, healthy diet and increased physical activity are considered the fundamental basis of a successful treatment . Higher protein intake, foods with a low glycemic index and lower fat consumption may help diabetic patients . Moreover, recent studies suggest that the intake of fresh fruits and vegetables is related with a lower incidence of T2DM. In a randomized controlled trial, it has been explained and demonstrated how diets rich in fruits and vegetables can improve blood glucose and insulin secretion . Furthermore, studies also indicate that bio-active compounds present in fruits, vegetables, spices, legumes, edible flowers, mushrooms, and medicinal plants, are prospective candidates for the prevention and the control of T2DM . Fruits and vegetables are rich sources of bio-active compounds, particularly polyphenols, exerting numerous positive human health effects. Polyphenols are plant secondary metabolites that act as a defense against pathogens, diseases, predators, ultraviolet radiation, parasites, and oxidants . Polyphenols are widely used in the food industry as a natural antioxidant ingredient in foods by their effects on preventing lipid oxidation and oxidative rancidity . Polyphenols are classified as flavonoids and non-flavonoids; flavonoids are divided in 12 groups including, flavonols, flavanols, flavons, flavan-3-ols, anthocyanins, flavanones, isoflavones, and dihydrochalcone. Non-flavonoids include phenolic acids, lignans and stilbenes. Following intake of polyphenols, they undergo important metabolism , which start in the enterocytes of small and large intestines before being absorbed into the circulation. Once in the hepatocytes, the hydroxyl group in the flavonoids undertake glucuronidation, methylation and sulfation to be available to enter in blood circulation. The next step is their flux to all organs and finally their elimination through urine. In the large intestine, the colonic microbiota will produce an extensive breakdown of the original polyphenolic structure into a phenolic metabolites with low-molecular-weight, this will produce a better absorbability of the compounds, and its passage to the secondary phase metabolism .Anthocyanins are hydrophilic pigments contained in red, violet, and blue colors of fruits and vegetables. Most abundant anthocyanins are glycosylated forms of cyanidin, delphinidin, malvidin, peonidin, petunidin, and pelargonidin. Anthocyanins are commonly linked to a sugar molecule, frequently glucose, nevertheless, rhamnose, galactose, and rutinose can also be present. Anthocyanins play an important role in obesity and diabetes prevention. Different studies have suggested that, once absorbed, anthocyanins can positively modulate GLUT4 in skeletal muscle and adipose tissue; other authors conclude that these phytochemicals may affect gastrointestinal microbiota and impact the host health . One of the foods rich in anthocyanins are BB.

For each generation, a 3-week treatment window minimized application costs by combining the insecticide with preventative treatments for powdery mildew. This information was widely disseminated via conference calls with government agencies, a UC electronic newsletter, industry associations’ e-news blasts, and grower liaisons in Napa, Sonoma, Fresno and Mendocino counties. An analysis of the PUR datafrom 2011 and 2012 shows a high level of compliance for treatment timing among Napa County growers . Mating disruption. Mating disruption programs deploy synthetic – 7,9-dodecadienyl acetate — the main component of the female sex pheromone — in hand-applied dispensers . When applied as an area-wide control strategy in Palearctic regions, MD has provided sustained control of EGVM populations while decreasing reliance on insecticides and reducing conflict between agricultural and urban populations . Although MD does not completely inhibit EGVM mating, delayed mating reduces populations because older females produce fewer eggs than younger females . TWG scientists strongly supported the use of MD as a control tool , large pots plastic and by the second EGVM flight of 2010, Isomate EGVM pheromone dispensers became widely used .

After the MD and insecticide programs were implemented, trap catches and visual inspections revealed dramatic decreases in population size in Napa County . Beginning in 2012, all Napa County vineyards within the defined treatment areas received pheromone dispensers through federal, state and local funding programs. Because MD limits the reliability of sex pheromone–baited traps and makes it very difficult to detect residual populations, the EGVM program has avoided the use of MD in treatment areas as they transition to deregulation. Urban and residential treatment programs. CDFA personnel used multiple strategies to manage EGVM populations in noncommercial grapevines in urban and residential areas. The organic product B. thuringiensis was applied during the first and second generations if the crop was to be harvested; if not, flower and/or fruit clusters were removed in the spring or early summer . MD was also used in certain areas and select counties. Alternate host surveys Polyphagy by EGVM has been documented in the literature: Larvae may feed on up to 40 hosts in 27 plant families . However, larvae are rarely found on hosts other than Daphne gnidium and Vitis; exceptions seem to result from adaptations to local climate and flora or elevated pest pressure and presence of ripe fruit .

UC and USDA personnel used pheromonebaited traps and visual surveys to monitor a variety of plant species in Napa County that are reported to be EGVM hosts ; no EGVM life stages were found during these surveys, suggesting that these species currently pose little risk in California. In separate surveys of olive orchards, 10 eggs and 12 EGVM larvae were found, indicating that olive flowers were a minor host in Napa during the first EGVM generation, but olive fruit did not host the second or third generations. The main host of consequence in California continues to be cultivated grape, Vitis vinifera.To prevent the movement of EGVM on regulated articles grown inside quarantine areas, CDFA required businesses to sign compliance agreements that mandated specific activities prior to and during harvest, transport, processing and waste handling. Compliance agreements were also required for similar activities in raisins and regulated fruit other than grape, including olive, persimmon, pomegranate, most stone fruit and specific caneberries. Based on recommendations provided by the TWG, in 2012 the USDA revised the list of regulated articles to exempt olive fruit and Rubus spp., and limited the acreage of Prunus spp. affected. Management of winery waste The possibility that EGVM could survive in unfermented winery waste was addressed by requiring that waste be composted on site, transported to an approved compost facility or returned to the vineyard of origin, depending on where the fruit was sourced relative to the quarantine area and county.

Alternatively, if grapes were pressed to a minimum of 2 bars or 28 psi, then movement of waste was unrestricted within California. Investigations provided evidence that EGVM larvae could survive on unpressed green waste after destemming and determined that in general the processing equipment was likely a greater source of contamination than unfermented waste . Mature larvae may move from clusters to protected locations, emphasizing the need for thorough washing of all equipment used to harvest, transport and process infested loads of grapes. Outreach and educational program EGVM program leaders provided transparent, consistent, timely and coordinated communication to parties directly and indirectly affected by the EGVM program. USDA led international communications and jointly with CDFA coordinated statewide communication. County agricultural commissioners and UC advisors directed local communication and assisted in other efforts. The outreach program also collected relevant information from stakeholders to ensure the appropriateness of regulatory requirements and adapted the program to local conditions and concerns. This openness and flexibility to change fostered the development of trust, respect and cooperation among all parties. The proactive, local campaign to communicate, educate and collaborate with a diverse community became a hallmark of the EGVM program . Industry outreach and education. Many methods were used to communicate information to growers and winery personnel. Educational materials included two tri-fold brochures, a poster and training videos developed and distributed through UC Cooperative Extension. Mass marketing campaigns raised public awareness using postcards, door-hangers, magnets, billboards and campaign signs, public service announcements, and online and social media; cross-linked websites provided comprehensive and current information. The online UC IPM Pest Alert presented information on pest identification, biology and management. Grower liaison/outreach coordinators in Napa, Sonoma, Mendocino and Fresno counties played an important role in outreach and educational efforts. In each county, the EGVM grower liaison conducted targeted outreach and was a trusted, independent source of information for growers and winery personnel. Program information was communicated at meetings, seminars and field days, as well as through individual communications and a UC Cooperative Extension Napa County newsletter. Farmworkers — particularly important “first responders” capable of identifying potential threats during their daily work — were reached through Spanish language presentations, field days and outreach materials. Growers also shared information and related their experiences through peer networks that strengthened the formal educational program. Public outreach and education. An estimated 2.5 million acres of California farmland are adjacent or in close proximity to nonfarm residences . To address potential areas of urban-agricultural conflict, county agricultural commissioners supported by USDA, CDFA, UC and industry groups worked within established local networks of community leaders to develop a public outreach campaign. County supervisors and city council members, environmental, square planter pots community and commercial organizations, residents and tourists were the targets of the outreach efforts, which built trust between program and community leaders, growers and the public. Program update The conditions set forth by the TWG specified that in areas attempting to qualify for deregulation no moths must be captured during five consecutive generations, insecticide treatments must continue to target the first and second generation, MD may not be used during the final two generations that the area is under regulation and during these two generations, trap density must increase to 100 traps per square mile  in all vineyards within 1,640 feet of previous detections.

Under California conditions, only the first and second are considered full generations because a proportion of the second generation enters diapause . Four counties were removed from regulation at the beginning of 2012, and by the end of 2012, five additional counties had been deregulated partially or in full. Napa County and portions of neighboring Sonoma and Solano counties remained under regulation in 2013, an area encompassing 686 square miles . Due to the historically large and widely distributed populations in Napa County, the TWG recommended a revised approach to deregulation: High-density traps deployed county-wide must be free from detections during four full flights before areas will be considered for deregulation . Any moth captured will trigger the delimitation and establishment of treatment areas within 1,640 feet of the detection. Consequently, trap density nearly tripled in Napa County in 2013 and the area under MD shrank from 23,000 to 2,800 acres in 2013 and approximately 1,907 acres in 2014. The remaining EGVM population in Napa County has been drastically reduced in size and distribution . By mid-August 2014, southern Sonoma County, the remainder of Solano County, and a portion of southern Napa County were removed from quarantine. The remainder of Napa County and a portion of northern Sonoma County, an area totaling 446 square miles, will continue to be regulated in 2015. Pheromone-baited trap surveys continue in other grape-growing areas of California and the United States as part of CDFA/USDA early detection programs. These surveys detected one moth in Sonoma County in 2014; this was not sufficient to trigger establishment of a new quarantine boundary. A model collaborative effort Development and implementation of a successful regulatory program in response to the detection of an invasive species in California agricultural systems requires a concerted and coordinated effort to address the pest while balancing the needs of the agricultural industry, trading partners and the general public. Regulatory, fiscal, environmental and biological aspects must be weighed to develop goals and determine program activities. The leaders of the California EGVM program were sensitive to these issues and gained credibility and trust by involving a diverse community for dialogue, responding to the needs of local communities, considering the impact on the environment, adopting new scientific information, investing in relationships and networks, and ensuring the appropriateness of regulatory requirements . Factors such as the insect’s limited host range, which allowed the program to focus on commercial vineyards; the use of management tools that minimized the impact to non-target organisms and are compatible with organic and backyard vineyards; and the long history of EGVM management in the Palearctic region also contributed to the successes of the program. The EGVM program leaders were recognized with the USDA Administrator’s Award in September 2012, acknowledging the program’s successful collaborative approach. The program has clear direction and well-defined goals, is grounded in biology, engages a diverse community and is responsive to the changing needs of participants. The participation of USDA, CDFA, agricultural commissioners, UC and other university scientists, growers, industry groups, community leaders and the general public resulted in a model effort that has reduced EGVM populations to a few areas of Napa and Sonoma counties.Variation in plant reproduction is central to processes from forest dynamics to farmer livelihoods. In perennial plants, masting marks one extreme end of the spectrum of population-level variation, and constant yield marks the opposite end. To date, much of the research on synchronous seed production has been focused on mast-seeding by wind-pollinated trees in temperate regions. It may be that mast-seeding is more common in wind-pollinated taxa; theory suggests selection for enhanced pollination efficiency through synchronous flowering with conspecifics is more likely in wind-pollinated species. In insect-pollinated species, synchronous flowering may saturate insect pollinators and high pollination efficiency at low flowering density may select for a more constant production of flowers . An alternative explanation of the over representation of wind-pollinated species in the synchronous seeding literature is that much of the masting work, and indeed the bulk of ecological and evolutionary research, has been done in temperate regions where wind is the predominant pollination syndrome among forest trees. Early reviews on whether pollination syndrome predicts the tendency for masting had difficulty gathering sufficient data on insect pollinated and animal-dispersed taxa. A recent meta-analysis included data with approximately equal numbers of animal- and wind pollinated species, but there were more time series per species for the wind- than animal-pollinated ones . To help to fill this gap, we make use of an analogous pattern of highly variable reproduction in perennial crop plants which, unlike mast-seeding forest trees, are biased toward insect-pollinated taxa and span tropical, Mediterranean and temperate climates . Alternate bearing in fruit and nut crops is an intermediate pattern of perennial reproductive variability in which a year of high reproduction is followed by a year of low reproduction. While media and trade reports have cited alternate bearing in discussions of national crop yield, literature on the extent and drivers of synchrony among alternate-bearing individuals is scarce. Despite evidence of similar plant-level mechanisms in masting and alternate bearing, ecological research on the synchrony of mast-seeding has largely ignored, or explicitly excluded, alternate-bearing crops .

Cabernet Sauvignon, Sauvignon Blanc and Cabernet Franc are characterized by specific volatile thiols and methoxypyrazines. Enzymes involved in the production of these aromas have been recently characterized. Phenolic compounds play a central role in the physical mouthfeel properties of red wine; recent work relates quality with tannin levels. While the grape genotype has a tremendous impact on tannin content, the environment also plays a very large role in grape composition. The pathway for phenolic biosynthesis is well known, but the mechanisms of environmental influence are poorly understood. Ultimately, there is an interaction between molecular genetics and the environment. Flavor is influenced by climate, topography and viticultural practices. For example, water deficit alters gene expression of enzymes involved in aroma biosynthesis in grapes, which is genotype dependent, plastic gutter and may lead to increased levels of compounds, such as terpenes and hexyl acetate, that contribute to fruity volatile aromas. The grapevine berry can be subdivided into the skin, pulp and seeds.

A thick waxy cuticle covers the epidermis. The hypodermal cells contain chloroplasts, which lose their chlorophyll at veraison and become modified plastids; they are the sites of terpenoid biosynthesis and carotenoid catabolism. Anthocyanins and tannins accumulate in the vacuoles of hypodermal cells . Pulp cells are the main contributors to the sugar and organic acid content of the berries. Pulp cells also have a much higher set of transcripts involved in carbohydrate metabolism, but a lower set of transcripts involved in lipid, amino acid, vitamin, nitrogen and sulfur metabolism than in the skins. Hormones can influence berry development and ripening. Concentrations of auxin, cytokinins and gibberellins tend to increase in early fruit development of the first stage. At veraison, these hormone concentrations have declined concomitant with a peak in abscisic acid concentration just before veraison. Auxin prolongs the Stage 2 lag phase and inhibits anthocyanin biosynthesis and color development in Stage 3. Grapevine, a nonclimacteric fruit, is not very sensitive to ethylene; however, ethylene appears to be necessary for normal fruit ripening.

Ethylene concentration is highest at anthesis, but declines to low levels upon fruit set; ethylene concentrations rise slightly thereafter and peak just before veraison then decline to low levels by maturity. Ethylene also plays a role in the ripening of another non-climacteric fruit, strawberry. ABA also appears to be important in grape berry ripening during veraison when ABA concentrations increase resulting in increased expression of anthocyanin biosynthetic genes and anthocyanin accumulation in the skin. ABA induces ABF2, a transcription factor that affects berry ripening by stimulating berry softening and phenylpropanoid accumulation. In addition, ABA affects sugar accumulation in ripening berries by stimulating acid invertase activity and the induction of sugar transporters. It is not clear whether ABA directly affects flavor volatiles , but there could be indirect effects due to competition for common precursors in the carotenoid pathway. Many grape berry ripening studies have focused on targeted sampling over a broad range of berry development stages, but generally with an emphasis around veraison, when berry ripening is considered to begin. In thisstudy, a narrower focus is taken on the late ripening stages where many berry flavors are known to develop in the skin. We show that that the abundance of transcripts involved in ethylene signaling is increased along with those associated with terpenoid and fatty acid metabolism, particularly in the skin.

Cabernet Sauvignon clusters were harvested in 2008 from a commercial vineyard in Paso Robles, California at various times after veraison with a focus on targeting °Brix levels near maturity. Dates and metabolic details that establish the developmental state of the berries at each harvest are presented in Additional file 1. Berries advanced by harvest date with the typical developmental changes for Cabernet Sauvignon: decreases in titratable acidity and 2- isobutyl-3-methoxypyrazine concentrations and increases in sugar and color . Transcriptomic analysis focused on four harvest dates having average cluster °Brix levels of 22.6, 23.2, 25.0 and 36.7. Wines made in an earlier study from grapes harvested at comparable levels of sugars or total soluble solids to those in the present study showed clear sensory differences. Six biological replicates, comprising two clusters each, were separated into skins and pulp in preparation for RNA extraction and transcriptomic analysis using the NimbleGen Grape Whole-Genome Microarray. Thus, a 4 × 2 factorial experimental design was established. After standard microarray processing and data normalization, two-way ANOVA indicated that the transcript abundance of 16,280 transcripts statistically significantly changed across the °Brix levels below the adjusted p-value of 0.05 , the transcript abundance of 10,581 transcripts changed significantly across Tissue types, and the abundance of 2053 transcripts changed significantly with respect to the °Brix x Tissue interaction term p-value column: adjBrix, adjTissue or adjTissue*Brix. A note of caution must be added here. There are high similarities amongst members in certain Vitis gene families , making it very likely that cross-hybridization can occur with probes on the microarray with high similarity to other genes. We estimate approximately 13,000 genes have the potential for cross-hybridization, with at least one probe of a set of four unique probes for that gene on the microarray potentially cross-hybridizing with probes for another gene on the microarray. Genes with the potential for cross hybridization have been identified and are highlighted in light red in Additional file 2. The rationale to include them is that although individual genes can not be uniquely separated, the probe sets can identify a gene and its highly similar gene family members, thus, providing some useful information about the biological responses of the plant. An additional approach was taken, removing cross-hybridizing probes before quantitative data analysis . Many of the significant genes were unaffected by this processing, but 3600 genes were completely removed from the analysis. Thus, it was felt that valuable information was lost using such a stringent approach. The less stringent approach allowing for analysis of genes with potential cross hybridization was used here in the rest of the analyses. To assess the main processes affected by these treatments, the gene ontologies of significantly affected transcripts were analyzed for statistical significance using BinGO. Based on transcripts that had significant changes in abundance with °Brix level, 230 biological processes were significantly over represented in this group . The three top over represented processes were response to abiotic stress, biosynthetic process, and response to chemical stimulus, a rather generic set of categories. Tissue differences were more revealing at the stage when flavors peak; 4865 transcripts that were significantly higher in skins compared to pulp at 23.2 °Brix were tested for over represented GO functional categories . Some of the top GO categories included photosynthesis, isoprenoid biosynthesis, and pigment biosynthesis . Some of the transcripts with the largest differences between skin and pulp at 23.2 °Brix are β-ketoacyl-CoA synthase , taxane 10-β-hydroxylase , wax synthase, a lipase, an ABC transporter, blueberry container and phenylalanine ammonia-lyase . The abundance of 5716 transcripts was significantly higher in pulp than skin at 23.2 °Brix . Some of the top GO categories over represented were a variety of transport processes and small GTPase mediated signal transduction . Some of the transcripts with the largest differences in abundance with pulp greater than skin at 23.2 °Brix were polygalacturonase , flavonol synthase, stachyose synthase, an amino acid transporter, a potassium channel , and HRE2 . The transcript abundance of 2053 genes had significantly differential expression across °Brix levels and tissues .

The top GOcategories over represented in this set involved photosynthesis and phenylpropanoid metabolism, both associated with the berry skin . Other flavorcentric categories of the 57 categories over represented include aromatic compound biosynthesis, fatty acid metabolism and alcohol catabolism. This transcript set was further analyzed by dividing into 10 clusters using k-means clustering . The over represented GO categories were determined for each cluster . Eight of the 10 clusters had distinct over represented GO categories; two clusters did not have any over represented GO categories, meaning that the genes in these two clusters were assigned to GO categories of expected proportions when compared to the entire NimbleGen array. Clusters 1, 8, 9 and 10 had a large number of over represented categories. Many GO categories within a cluster are subsets of others in that cluster and were grouped together. For example, cluster 4 had four over represented GO categories, oxygen transport, gas transport, heat acclimation and response to heat. The four categories could be grouped into two, as two are subsets of the others; this is how they were listed in Table 1.It would be impossible to discuss here all the transcript abundance changes detected in these berries. As we were interested in compounds associated with berry flavors as they develop or change in the late stages of berry ripening, we took a more targeted approach for analysis with this in mind. Berries at 24° Brix are known to be near-optimal for flavor, thus we took a simple approach to look for genes that were peaking around this stage. We found some significant and large increases in transcript abundance between the 22.6 and 23.2 °Brix levels. A group of VviERF6 transcription factor paralogs represented 6of the top 10 transcripts increasing in transcript abundance from 22.6 to 23.2 °Brix in the skin, but not in the pulp . These VviERF6 TFs were also found in Cluster 8 . This is very interesting since many flavor compounds are derived from the skin and ERF TFs are known to be responsive to ethylene, a known fruit-ripening hormone. These VviERF TFs were named ERF105 in the annotation by Grimplet et al. , however they are more orthologous with AtERF6 as determined by a more comprehensive phylogenetic method using many plant species at Gramene . Annotation details of the V1 gene models of the VviAP2/ERF superfamily can be found in Additional file 8 including updated Vvi symbols according to its closest Arabidopsis ortholog as instructed by the Grapevine Gene Nomenclature System developed by the International Grape Genome Program Supernomenclature committee. This renaming of the AP2/ERF superfamily should facilitate comparative analyses and functions with other species, particularly Arabidopsis. To properly annotate the AP2/ERF superfamily of Vitis vinifera according to the IGGP Supernomenclature committee instructions, a phylogenetic tree was generated for the AP2/ERF superfamily of Arabidopsis thaliana and Vitis vinifera using the TAIR 10 and V1 gene models, respectively . The labeled family classifications were derived from the Arabidopsis naming scheme by Nakano et al.. There are 130 members in the VitisAP2/ERF superfamily in the Pinot Noir reference genome. However, the six paralogs of ERF6 discussed above belong to a Vitis vinifera clade in subfamily IX and are distinctly different or separate from any Arabidopsis subfamily IX ERF TFs . Using k-means clustering, VviERF6L1 fell within Cluster 8 with 369 transcripts, including five additional VviERF6 paralogs. The top GO categories associated with Cluster 8 were genes associated with terpenoid metabolism and pigment biosynthesis . Other interesting flavor associated categories included fatty acid and alcohol metabolism . Representative transcripts from Cluster 8 that were correlated with the transcript abundance profile of VviERF6L1 can be seen in Figure 4. These are ACC oxidase, which is involved in ethylene biosynthesis; a lipoxygenase, part of a fatty acid degradation pathway giving rise to flavor alcohols such as hexenol; α-expansin 1, a cell wall loosening enzyme involved in fruit softening, and two terpene synthases, which produce important terpenes that contribute to Cabernet Sauvignon flavor and aroma. The high similarity of these transcript profiles indicates that ethylene biosynthesis and signaling may be involved in the production of grape aroma. Supporting this argument, two recent studies have shown that a tomato ERF TF , falling in the same ERF IX subfamily, has a strong effect on ethylene signaling and fruit ripening. The transcript abundance of AtERF6 in Arabidopsis is strongly increased by ethylene, which is triggered by the MKK9/MPK3/MPK6 pathway. The transcript abundance of VviMKK9 in the Cabernet Sauvignon berries was higher in the skin than the pulp, but there were no significant differences for VviMPK3 or VviMPK6 . This is not too surprising since AtMKK9 activates AtMPK3 and AtMPK6 by phosphorylation. In addition, the transcript abundance of AtERF6 in Arabidopsis increases with ROS, SA, cold, pathogens, and water deficit. There were no visible signs of pathogen infection in these berries. Additional circumstantial evidence for ethylene signaling in the late stages of berry ripening was that the transcript abundance of many VviERF TFs was significantly affected by berry ripening and/or tissue .

Shannon diversity and evenness were fit with Gaussian distributions while all other variables were fit with Poisson distributions. In comparing the ratios of viable seeds to total seeds vs. the ratio of viable seeds to counted stigmas, we found that there was a strong correlation between these metrics. To look at the effect of land use type on seed-set, we therefore decided to utilize the ratio of viable seeds to total seeds in each seed head that did not experience seed predation, because of error in counting the number of stigmas . We then used a generalized linear mixed model fit with a Binomial distribution, with land use type as a fixed effect and site as a random effect. Finally, we tested for an effect of floral visitor observations on yellow starthistle seed set at each site. We averaged the number of visits from each morphotype across temporal observation events at the same site. Morphotypes that averaged at least one visit per 30 minute observation window were included as fixed effects in a linear mixed model fit with a binomial distribution, with site as a random effect and the ratio of viable to total seeds as the response variable. We also modeled the effects of total bee visitation, morphotype richness, and morphotype diversity on seed set ratios.Our results show that rates of bee visitation and seed set vary among urban, agricultural, square plastic plant pot and natural landscapes, demonstrating the importance of land use in the dynamics of plant pollinator interactions.

We suggest that these effects are at least in part explained by floral availability, a vital bee resource, which can be highly variable among different land use types. For example, in August there are few plants in flower besides yellow starthistle in the natural areas of Contra Costa County, California, whereas in urban and agricultural areas there are many exotic plants and supplementary inputs available . From pan-trapping of bee specimens in the region , we know that total bee abundance is highest in the spring in natural areas. However, towards the end of the summer when yellow starthistle is in flower, there is little difference in collected bee abundance between human altered landscapes and natural areas, and human-altered areas may even exhibit overall higher bee abundance. Our results of bee visitation to yellow starthistle support this pattern. Agricultural areas have large populations of managed honey bee colonies, so one would predict visitation to yellow starthistle by honey bees to be positively associated with surrounding agricultural land use. By contrast for native bees , the highest rates of visitation to yellow starthistle were in sites with more surrounding urban land use. Urban gardens have many exotic plants, often selected for aesthetic purposes, many of which are in flower later in the season than most California native plants. In addition, many of the plants in urban areas both directly and indirectly receive supplementary resources, particularly water, that further extend their flowering time.

Even though agricultural areas also have supplementary resources, the main crop in flower in East Contra Costa County later in the season is maize, which is wind pollinated. There may be multiple impacts of exotic plants in urban areas. By filling the phenological flowering gap noted above, they may help attract even larger populations of bees into the urban landscape. In addition, bees in urban sites may be behaviorally more likely to visit non-native plants due to the increased encounters they have with novel plants.In agricultural and natural landscapes, a positive correlation between pollinator visitation and seed set is typical. Surprisingly in our system, in human-altered landscapes, higher total observed bee visitation did not result in higher proportions of seed set, as would be expected. In fact, urban areas, despite receiving the highest rates of native bee visitation, exhibited the lowest rates of seed set. Conversely, natural areas, which received the lowest amount of total bee visitation, had the highest rates of seed set. We suggest 2 possible explanations for this discrepancy between pollinator visitation and rates of seed set: 1) pollinator efficiency; and/or 2) the composition of the local flowering community. Depending on the plant, certain pollinator species are much more effective than others. For example, Osmia, Habropoda, and Apis, have been found to produce varying amounts of seed set as a result of a single visit to blueberry, but these results vary slightly depending on the blueberry variety. In the case of yellow starthistle, it is likely that the most frequent visitors are perhaps not the most efficient. When we directly compared average seed set at each site against visitation rates, we found a significant positive association with the medium hairy leg bees.

The medium hairy leg bee morphotype includes those species which fall in both the Tribes Emphorini and Eucerini. Emphorini are known to largely be oligolectic , meaning they specialize on certain plant groups, which theory suggests would make them more efficient pollinators than generalists. It was also the only group that was observed most frequently during morning sampling, perhaps reflecting a difference in when yellow starthistle is most receptive to pollination. Despite the overwhelming abundance of honey bees in agriculture areas, we did not observe higher seed set in those regions, consistent with the observation that honey bees can be poorer pollinators than other species. It is also important to note that this study used a morphotype classification, and there may be multiple species that fit within the same morphotype that provide varying degrees of pollination services. It is possible there are rare, but highly efficient, pollinators that were rarely observed during the sampling period, or were lumped together with a more frequently observed morphotype. An alternative explanation for the lack of an association between floral visitation and seed set is that higher plant diversity in urban and agricultural areas may decrease pollinator efficiency. Previous research has shown that invasive alien plants can have a negative effect on native plant communities by acting as attractors for pollinators, or decreasing pollinator efficiency by providing a wider range of resources for pollinators to visit, with the consequence that visitors transfer pollen from non con-specifics, potentially clogging stigmas and reducing pollination success. In this case, our target plant, yellow starthistle is indeed considered an invasive alien plant, but the hypothesis of it being in a novel diverse community could lead to a similar effect on the frequency and quality of pollination services that it receives. In sites where there are many other potential plants to visit and accompanying decreased floral fidelity leading to diverse pollen loads, one predicts decreased pollinator efficiency. Abundant sources of exotic plant pollen could occur in areas where there is a greater diversity of nearby plants for pollinators to visit. This explanation might account for the observation that shield tipped small dark bees were negatively correlated with seed set. We selected yellow starthistle as the target plant for this study because of its ubiquitous distribution, reliance on pollination, 25 liter square pot and its attraction for a wide set of visitors; it is also a highly invasive and undesirable plant. Previous research on yellow starthistle has found that itsinvasion can be facilitated other non-native pollinator species such as the honey bee, Apis mellifera, and the starthistle bee, Megachile apicalis, which is included in the medium striped hairy belly bee morphotype. However, the abundance of bees in both of these 2 morphotypes were most closely associated with agricultural areas, which did not have the highest rates of seed set as would be predicted by visitation alone. Our results indicate clearly that bee visitation in human-altered landscapes can be higher than that in comparable natural areas, especially towards the end of the flowering season when there are few resources available in natural landscapes. Because the response of bee visitors to land use change depends on species-specific requirements and these pollinators also have variable effects on plants, understanding the effect of land use change on pollination services requires knowledge not only of which pollinator groups shift to the human-altered landscapes, but also the rate of pollination that those groups have on the plant species in those landscapes. Future research will benefit from looking at a wider range of plants with a different range of target pollinators and that flower earlier in the year to better tease out these hypotheses.

If the patterns of bee visitation and seed set that we observed are indeed consistent across other plant species, the novel plant communities created in these human-altered landscapes and the generalist bee species that are favored in such landscapes will lead to a reduction in overall pollination services. As a viscous soluble fiber, pectin is associated with lowering blood serum total cholesterol and low-density lipoprotein cholesterol , without changing high-density lipoprotein cholesterol . By increasing satiety, pectin can help weight control by reducing food consumption. . Due to its complex structure, pectin is a common drug formulation agent for oral delivery, in the form of tablets, gels, hydrogels, beads, aerogels, coated, and compression-coated dosage . It is especially beneficial for colonspecific drugs as the bio-active compounds, mostly short-chain fatty acids, and can be released after hydrolyzation of pectinolytic enzymes and microorganisms in the colon . The rhamnogalacturonan type I fragments within the pectin also have lower molecular weight to function as an anticancer down-regulator of galectin-3, which is a pro-metastatic protein in many cancer cells. Pectin has also been used to nano-encapsulate bio-active compounds, thereby enhancing shelf life and stability.Pectin is considered a safe food substance through various government food agencies. Even though its usage and composition are regulated under different food additive laws, the Food and Drug Administration of the U.S. considers it GRAS. At the Joint FAO/WHO Expert Committee Report on Food Additives and in the European Union, no numerical acceptable daily intake has been set, as pectin is considered safe. Its abundant benefits stimulated great demands and the global pectin market size was valued at USD 964.1 million in 2015 .Polyphenol compounds can bind with the cell wall polysaccharides to strengthen the structure . In return, CPSs can interact with polyphenols to modify their bio-accessibility, bio-availability, and bio-efficacy . Liu et al., summarized multiple research that has been done to investigate the binding effects between pectin and polyphenol. Noncovalent bonds are dominant in the polyphenol-pectin interactions, such as hydrogen bonds, hydrophobic interactions, electrostatic interactions. These weak bonds are sensitive to the intrinsic properties of polyphenol and pectin , as well as environmental conditions .Considering the binding property of pectin and polyphenols, a co-extraction of pectin and polyphenol should be considered because of the following advantages: Boosted stability of polyphenol and overall antioxidant activity; and Enhanced unique and beneficial characteristics for developing novel food products. As for stability, Oszmiański et al. proposed a cooperative hydrogen bond or hydrophobic interactions existed between the oxygen atom of pectin and the phenolic hydroxyl group. A natural pectin coating is speculated as less loss of polyphenols was observed due to the limited oxygen contact. The strength of pectin-polyphenol interaction is mainly determined by the non-covalent bonds, including hydrogen bonds, ionic bonds, and hydrophobic forces. Regarding the quality improvement, Hayashi et al., found the astringency of gallate-type polyphenol was reduced by the addition of pectin. A similar finding was reported in persimmons .Pectin polymer degradation is a fundamental step of extraction. The common degradation approaches include chemical hydrolysis, heating, radiation, and enzymatic reaction. Acid hydrolysis on pectin was one of the most conventional chemical degradation methods. Thibault et al. studied the mild acid hydrolysis of apple, beet, and citrus pectin. Inorganic solvents, including HCl and HNO3, have been widely used in the industry. However, they raised concerns in food applications and were difficult to recover, thus causing undesired pollution and high processing cost . Novel technologies with reduced pollution are in demand.Physical technologies, such as the non-thermal process, were superior due to their high efficiency, reduced pollution and cost, and ease of control. Ultrasound as a novel non-thermal technology has been widely applied in food industries, including extraction, preservation, emulsification, filtration, and cutting . Successful ultrasound applications in pectin extraction include citrus , apple , and sweet potato . Ultrasound extraction combined with acid hydrolysis was reported to further increase the pectin yield. Muñoz-Almagro et al. studied the pectin degradation under ultrasound power of 400 W in the presence of nitric acid and citric acid .

Diversify garden plantings Different types of pollinators have preferences for the plant type or flower structure that they visit to collect nectar or pollen resources. For example, hummingbirds have long beaks and tongues that can access nectar that other pollinators cannot. Additionally, other pollinators such as bats and moths visit flowers that are primarily open at night. Having a large diversity of plants ensures that pollen and nectar are accessible for a variety of pollinators. Some pollinators, like bees, require both nectar and pollen from flowers to meet their nutritional needs, whereas hummingbirds need a constant supply of nectar. A large variety of flower resource types allows for greater pollinator diversity.Use native plants Many pollinators have tightly interwoven relationships to native flowering plants. Pollinator emergence and life cycles are often synchronized with their preferred plants’ flowering patterns. It has been documented that native bees forage on native plants more frequently than on non-native plants . Native plants are also well adapted to the state or region’s localized climate and soil conditions.

Native California desert plants, such as desert globemallow , black plastic plant pots may be native to hot deserts; however, they thrive in cooler, coastal parts of the state as well, and the mallow attracts native bees wherever it is planted. Consider leaving flowering weeds Many plants that are considered weeds are actually attractive to pollinators. Leave some areas of the garden weedy for use by pollinators. This area can also serve as a bare-soil nesting area for native bees. Remove weeds immediately after flowering begins to decline to discourage them from setting seed or becoming invasive. Some weeds that are regularly used by pollinators include thistles , bristly ox tongue , and dandelions . Provide water Some pollinators, such as honey bees, need water for their survival. Maintain a shallow dish or bird bath with a landing surface to keep pollinators hydrated. Two methods to prevent honey bees from drowning are to place pebbles or stones in the dish or use a piece of floating cork board or wood. Some bee pollinators, including the blue orchard bee , require mud for nest construction . Provide shelter All pollinators need a protected place to either raise their young, lay eggs, or hide from predators. Types of cover include bird houses, bat houses, native bee boards or boxes, trees, shrubs, grasses or weeds .Avoid pesticides Pesticides not only kill the desired pest in your yard, but they can also harm valuable pollinators and beneficial insects that can provide pest control, given the chance.

Using pesticides in your garden may do more harm to your environment, including your pets and you, than what you might gain from using them. Avoid pesticides whenever possible, and use only cultural or biological controls to control the target pest. Also consider removing problematic plants and replacing them with another plant type. Refer to the University of California Integrated Pest Management website for information about controlling home garden pests safely by using the least toxic methods. Another resource is the brochure on pollinator-friendly pest solutions found on the Pollinator Partnership website .Recent news media has attracted public attention to honey bees since they have begun to decline under a series of pressures, including colony collapse disorder . Although native bees are not affected by CCD, they are susceptible to population decline from their own natural enemies and human disturbances. A growing public concern is currently about bee conservation. This new awareness, coupled with education surrounding native bees, has gardeners searching for ways to provide habitat. Honey bees are the most recognized and abundant bee in most gardens. Domesticated honey bees are generalized foragers, meaning that they visit a wide variety of host flowers. Native bees have more specific relationships to plant types and groups; they come in an array of colors, sizes, and shapes, and exhibit a wide range of fascinating foraging and nesting behaviors.

These predictable relationships between native bees and their flowers are the basis for planning a successful bee habitat garden. Select plants from the recommended plant list that attract both honey bees and a variety of native bees. A bee garden has four main components: 1) preferred plants, 2) single-flower types grouped into individual patches, 3) seasonal sequence of nectar and pollen , and 4) at least 20 plant types. A water source is necessary for honey bees but not for native bees. A highly variable habitat garden should start with at least 20 plant types to provide resources for a variety of bee species throughout the year. An ideal bee garden would have at least 20 patches of flowering plants with each plant type in its own patch. To select plants, consult table 1 as well as recommended plant lists from recognized online resources, such as the UC Berkeley Urban Bee Lab website . It is important to select a variety of plants with nectar or pollen, as they are both necessary to meet the nutritional needs of native bees. Native bees have taxonomic groups with distinct emergence times that can be matched with their preferred floral hosts. Native bees have four distinct foraging seasons, while honey bees continue to forage through all four seasons . For example, early spring bees emerge in February to visit plants such as Ceanothus species and Arctostaphylos species . Selecting plants to match multiple bees’ foraging seasons will encourage diverse groups of bees to visit your garden. Through careful placement of plants, one can increase flower foraging for bee visitors. Arranging plants in patches of one plant type, with patches measuring at least 4 by 4 feet, will catch bees’ attention as they search for flowers. Patches encourage bees to forage longer, and they make bees easier for gardeners to observe. Bees are always searching for new plants , which means that once the new habitat is completed they will be quick to appear.Soil-nesting bees Providing nesting resources for native bees transforms a habitat garden that allows bee populations to build up through time. Approximately 70 percent of native bees in California are solitary ground nesters, while 30 percent are cavity nesters. Ground-nesting bees look for patches of bare soil or soil with light leaf litter to construct nest tunnels to build their brood cells. Nesting is the least studied component of native bee ecology, and only general knowledge exists about the best soil type for native bees. Soil preferences vary from species to species, black plastic planting pots with preferences ranging from sandy soils to clay-based soils. The best approach to encourage bee nesting is to leave the entire yard or a sunny area mulch-free. Common gardening practices of heavy mulching create physical barriers that prevent native bees from nesting, so use mulch sparingly. Cavity-nesting bees Cavity-nesting bees include those that use existing holes or spaces in which to construct their nests. One can help these bees by providing artificial nest sites for them, which are easy to construct or purchase from commercial outlets. Most cavity-nesting bees prefer to use dead-end holes 4 to 6 inches deep and 3⁄16to 5⁄16inch in diameter. Hollow sticks, reeds, bamboo sections, and drilled holes in wood blocks meeting these measurements are suitable for use. Normally these are tied into bundles with wire and hung horizontally in a protected and warm, north-facing area. A tiny roof can be added for additional protection from rain. Note that exposure to direct sunlight for extended periods may overheat the nests and kill the brood. We also recommend cleaning or discarding nests after 2 years of use to prevent a buildup of parasites and diseases. On its website, the USDA Agricultural Research Service provides a wealthof information on techniques for constructing artificial nests and managing bees that use them .

One cavity-nesting bee, Osmia lignaria, commonly referred to as the blue orchard bee , has proven effective in pollinating crops such as apples, cherries, and almonds. This bee prefers 5⁄16-inchdiameter holes for its nest sites, but it also requires mud to construct the brood cells and close the entrance of completed nests. Growers and gardeners wishing to attract this bee can assist it by providing a nearby source of clay mud in a tub or trench . Besides the blue orchard bee, many other bees and beneficial predatory wasps may use the nests, providing a fascinating window into the little known world of garden ecology .A butterfly garden provides habitat for adult butterflies and their offspring to thrive . Host plant flowers are critically important to maintaining butterflies’ habitat. Providing preferred nectar-rich plants for adults is the starting point for providing habitat for these magnificent insects . It is important to plant various flowers that provide nectar in bloom from late summer into early fall when butterflies are most abundant. When adults visit a garden, they are not only looking for a sip of sweet nectar—they are also scoping out potential plant hosts upon which to lay their eggs. Butterflies do not feed and raise their offspring; once their eggs hatch, the larvae feed on the plant their mother has chosen for them until they are ready to pupate, transforming into a chrysalis. These eggs will be laid on specific host plants known instinctively by the adult to be a nutritious source of food for her caterpillar offspring. Caterpillars are messy eaters, leaving chewed-up leaves and droppings as evidence of their presence. While their feeding results in less than-picture-perfect plants, these host plants are a necessary food source for the growing caterpillar. If chewed leaves or flowers are undesirable, consider planting host plants in an out-of-the-way part of the garden .Plant attributes associated with hummingbirds include floral characteristics like a long trumpet shape, red color, large amounts of nectar, and little or no scent. Many tropical plants and at least 150 flowering plants in North America have evolved these characteristics to encourage hummingbird visitation . Hummingbirds use their uniquely shaped long bill and tongue to dip deep into flowers to access nectar that is inaccessible to most other pollinators. The flowers in turn have evolved to place pollen on the hummingbird’s head or body when it drinks nectar. Creating a garden with hummingbird plants, nesting habitat , and feeders will ensure visits from these fascinating birds. However, supplying them with a clean, nutritious food source requires weekly maintenance. Begin with a clean feeder and add a sugary solution that is one part sugar to four parts water. Using honey or a stronger sugar concentration could be harmful or even fatal. Adding red dye to the solution is discouraged; most hummingbird feeders have red parts that will catch the hummingbird’s eye. Feeders need to be taken down to be cleaned and refilled weekly. Wash the feeder with hot, soapy water, and rinse with hot, boiling water. Refill it with a fresh supply of solution, which can be made ahead and stored in the refrigerator. It is important to discard the old solution, as it can ferment into alcohol. If you live in a very hot area, the feeder should be cleaned and solution replaced every 2 to 3 days.The world’s population is predicted to reach nearly 11 billion by 2100; thus, sustainable agriculture and food safety are foremost issues . However, environmental pollution, increasing urbanization, and the gradual decrease in fertile soils have complicated these issues . It has become a critical priority to develop safe agricultural products to support environmental and human health. One innovative solution to the challenges presented by infertile soil and the need for water conservation may be soilless agriculture . Soilless agriculture is growing in popularity among commercial farmers because it eliminates soil-borne problems. The global market for hydroponic systems is estimated at $9.5 billion in 2020 and is predicted to reach $16.6 billion by 2025, growing at a five-year compound annual growth rate of 11.9% . The main strategy in soilless agriculture is to keep the system as clean as possible. However, a new trend is the inclusion of beneficial microorganisms to enhance the resistance to biotic and abiotic stress factors in the plants . Although beneficial microorganisms are found throughout the soil, they are most prevalent near plant roots in an area termed the rhizosphere . PGPRs are mostly obtained from the soil.

The recent growth in herbarium-based phenological research is arguably a product of the growing interest in climate change and phenology around the turn of 21st century . Researchers realized that herbarium specimens could potentially be used to detect and quantify long-term phenological shifts in response to climate change. This, in turn, led to the use of specimens to estimate phenological sensitivity to various environmental factors, including temperature, day length, and precipitation . To date, specimens have been used to estimate the onset of several phenophases, including first flowering, peak flowering, leaf-out, and fruit set, as well as the duration of entire growth phases. These phenophase estimates have been used to study long-term shifts in phenology and phenological sensitivity to interannual climate variation . A literature review focused on the modern use of herbarium specimens to study phenological responses to climate reveals interesting generalities and insights. First, drainage planter pot studies that have investigated longterm shifts in phenology have generally found that flowering and leaf-out times have advanced, in some cases dramatically, over the past century .

These long-term trends are often in agreement with studies that have used alternative sources, such as observational data, to study phenological shifts. Second, for most of the studies we reviewed the onset dates of spring flowering and leaf-out tended to be negatively associated with winter or spring temperatures ; that is, plants tended to flower and leaf-out earlier in warmer years. However, some species and regions exhibit delayed or mixed phenological responses under warmer temperatures, potentially because they did not experience sufficient winter chilling requirements or because the imprint of past climate conditions has resulted in a response lag. Third, given the span of time and geographic area that specimens encompass, they almost always capture a greater range of climatic variation experienced by a species than traditional long-term observational data, and thus can provide a more complete estimate of phenological shifts over time as well as phenological sensitivity to interannual or spatial variation in climate . Most studies that have used herbarium specimens have focused on a single phenological event, most commonly the date of onset for a single phenophase .

The most frequently studied phenophase in relation to climate change is flowering , with a specific focus on either mean flowering date or peak flowering date . Only a handful of studies have attempted to quantify different events within a phenophase, such as the onset, peak, and end of flowering date. Thus, the opportunities for expanded application of comparable and new techniques are abundant. For example, specimens can be used to assess multiple phenological characters at different stages of development , allowing researchers to estimate the sensitivity of different points in a given phenophase as well as determine how different phenophases are related. Additionally, most herbarium-based studies have been limited to northern, temperate biomes , mirroring geographic biases in long-term observational data. The potential to expand phenological investigation into nontemperate biomes using specimens, however, is considerable, as illustrated by the density of tropical and subtropical specimen records in the Integrated Digitized Biocollections database alone . Several recent studies have validated herbarium phenological estimates by comparing them with independent estimates of similar phenological phenomena . Generally, comparisons with independent phenological data – using photographs and field observations
show that herbarium-based estimates of both phenological timing and phenological sensitivity to climate are reliable . At a broader scale, additional validation of herbarium-based phenological data has come from comparisons with satellite observations of ‘green up’ .

While these studies provide important validation of herbarium-based phenological data, they are nonetheless limited in their phylogenetic scope and number of regional comparisons. As the use of herbarium-based phenological data grows, so too should efforts to independently validate these data.Herbarium-based data, like all sources of data, are subject to potential biases and limitations of which researchers must be aware. Such limitations are present from the specimen collection phase, to the digitization and processing of specimens, to the analysis and interpretation of specimen data. By understanding and addressing these challenges, researchers can make full and appropriate use of specimens for phenological research. Some limitations of using herbarium data for phenology are common to other observational datasets and originate at the time of specimen collection, including accurate species identification and phenological event and phase discrimination. While specimens are often correctly identified by experienced botanists, they may still be misidentified or labeled according to outdated taxonomy. Unlike with observational datasets, however, species and phenophase identifications for herbarium data can easily be confirmed by revisiting anomalous specimens.Herbarium data are known to contain additional, unique biases that stem from the opportunistic nature of their collection. Botanists often collect samples depending on their interests, schedule, and location and not to capture the phenological status of the plant perse. Collection biases relating to plant habit, morphology, and nativity may also occur in herbarium datasets; for example, Schmidt-Lebuhn et al. discovered strong biases against very small plants, plants with brown or green inflorescences, and introduced species in a sample of Australian Asteraceae. Rich and Woodruff noted that collections are biased towards common, showy plants that grow in clumps. Additionally, broader taxonomic, spatial, and temporal biases have been identified with Global Biodiversity Information Facility occurrence records, which include herbarium records . Specific to phenology, plants may be less likely to be collected at the very beginning or end of a reproductive season, especially if a species is difficult to identify during these stages or is inconspicuous. For example, Davis et al. found that first-flowering date estimates from specimens were, on average, 3 days later than first-flowering date estimates from field observations. Botanists may also collect only those individuals exhibiting a certain phenological stage to facilitate identification. However, it is also true that botanists may deliberately collect plants that are flowering or fruiting out of season and are therefore not representative of the overall phenology of the species. Another source of collection bias is the tendency for large numbers of specimens to be collected during single collecting trips, which can result in oversampling and the generation of duplicate specimens distributed to multiple institutions that are subsequently treated as independent samples. Duplication of records is a well-known problem, however, and efforts are currently underway to better account for duplicate records across databases and data portals. Finally, herbarium specimens often represent only a fragment of an entire plant , plant pot with drainage which makes it important to consider how accurately specimens represent the phenology of the whole plant or local population from which they are sampled.Data quality issues in herbarium data may also arise after collection, during label transcription, or due to digitization. For example, ambiguous handwriting or descriptions can lead to the incorrect transcription of a specimen’s location or collection date. In addition to transcriptionerrors, discriminating among phenophases can be even more difficult if observers are assessing digital images rather than the physical specimens themselves.

While these problems can often be resolved from other contextual clues , each of these aspects of data quality must be assessed and managed when studying phenology. Moreover, different countries and individuals have developed separate methods for recording specimen information, which presents a challenge for data aggregation. This topic has recently received renewed attention and methods to improve the standardization and integration of these data are currently being developed .Clearly, herbarium records are subject to error, as are all sources of data, and may contain geographic, phylogenetic, temporal, or other biases because they were not assembled to answer phenological questions. Nevertheless, one of the strengths of herbarium data is that their biases can be minimized by careful selection of species and phenological phases for assessment, rigorous training of observers, high-quality imaging, and the continued development of statistical methods to test and correct for biases.In the phenological studies that have been completed to date , researchers often evaluated phenological stages differently according to their research priorities and rarely madedata publicly available, thus limiting the utility of those data beyond the life of the individual projects. The most serious challenge for the future of herbarium-based phenological research is the standardization of phenological terms and methods for scoring phenophases and phenological events. Such standardization is important not only to ensure that herbarium-based studies are comparable but also to facilitate effective integration with other types of phenological data such as citizen science observations, satellite imagery, and stationary camera images. Biodiversity data standards for the biocollections community have already been established in the Darwin Core Data Standards. Most digitizing institutions generate data conforming to the Darwin Core, which comprises defined metadata properties and a small set of classes; however, phenological terms are not currently defined by the Darwin Core and instead are captured in unrelated fields such as ‘occurrenceRemarks’, ‘organismRemarks’, ‘dynamicProperties’, or ‘fieldNotes’. Many institutions capture flowering information in the ‘reproductiveCondition’ field, but this field lacks a standardized vocabulary. For example, we discovered 3900 unique terms to describe reproductive status in a search of the ‘reproductive Condition’ field of 5.7 million specimens in SEINet, a portal of digitized specimens for Arizona and New Mexico, USA. Lack of standardization complicates data integration and presents a huge obstacle to the mobilization and consolidation of herbarium data from multiple institutions for phenological research. The development of standards and ontologies is a vital step toward unlocking the research potential of digitized specimens. Standardization of herbarium specimen data, in combination with the availability of new data management tools, will facilitate the large-scale collection and use of phenological data from specimens. The task of scoring phenological data from millions of digitized specimens, however, is a monumental task. As noted above, herbarium-based phenological studies to date have typically focused on only a single phenophase and classified specimens in binary terms . This limited approach is due in no small part to the challenge of scoring phenology for a large number of specimens. Standardization can facilitate the collection of these data in two ways: by providing a template for scoring phenology that can be easily incorporated into the digitization or post-digitization workflow; and by providing guidelines for the conversion of raw count data collected via citizen science crowd sourcing into predefined phenophases.Efforts to scale up the collection of phenological data using new tools are already under way and would only benefit from the incorporation of a standardized ontology and data structure. The New England Vascular Plant project, for instance, has developed an extension of the specimen management system Symbiota that provides an interactive online platform to score a range of predefined phenophases based on coarse estimates of different phenological characteristics . This approach has the advantage of speed and efficiency and can be easily incorporated into an existing digitization pipeline where, along with transcribing the label information, technicians input phenological scores. Another tool, similarly meant to be implemented within an existing collection database, is the Phenological Predictability Index module in the Botanical Research and Herbarium Management System. The PPI module, however, is geared more toward standardizing estimates of phenological activity as opposed to scaling the collection of the data itself. Another avenue for scaling phenological data collection is the use of citizen science crowdsourcing. The popular citizen science platform Zooniverse has utilized crowdsourcing in the collection of data from digital specimens including label transcription [Notes from Nature ] and even phenological data [Orchid Observers ]. Another crowd sourcing tool that has been developed to collect phenological data from specimens is CrowdCurio. Preliminary results from CrowdCurio have demonstrated that phenological data collected from non-expert users are comparable to those compiled by expert users, suggesting that it has the potential to be a powerful tool for the collection of detailed, accurate phenological data. In addition to crowd sourcing, machine learning – the ability of computers to learn a task without being specifically programmed – offers an exciting new tool for the collection of large amounts of phenological data from specimens. Several recent studies have demonstrated that machine learning can be used to identify species with a high degree of accuracy based on leaf shape and venation. In either case data collected with these new and powerful tools should be made to conform to standardization efforts so that they can be easily incorporated into existing herbarium databases.