Horticulture

It was for the first time proved by National Institute of Allergy and Infectious Diseases that significant immunogenicity can be induced safely by an edible vaccine. After two years, antigens of Norwalk virus were expressed in potato .The nor walk virus antigens were expressed in transgenic tomatoes as well. Similarly,rabies glycoprotein and hepatitis B surface antigen were expressed in spinach and lettuce respectively. Plant as a bioreactor is cheaper, easy-to-handle, requires no sophisticated or cold storage facilities, easy to scale up, cost-effective or less-expensive production, involves effective, convenient and easy route of administration. Bacterial, viral, parasitic and immune-contraceptive vaccines can be produced in plants as edible vaccine .The first chimeric gene expressed in plants was human growth hormone expressed in sunflower and tobacco plants, transformed through Agrobacterium-mediated transformation.

First generation plant-based vaccines were produced against influenza virus, human papilloma virus and norovirus by modifying PVX or TMV. In second generation plant-based vaccines, deconstructed viral vectors devoid of different viral elements needed for its replication and infectivity were used. Recombinant viral vectors with heterologous coat protein, with cell surface presentation of foreign antigen in the viral coat protein and sub-genomic promoters etc were more stable, environmentally safe and provided high yield.Potato and tobacco were used as model organisms initially in development of plant-based vaccines. Potato was used as model plant in edible vaccine production since it was easy and efficient to transform, tuber-specific promoters could be used to express transgene/antigen gene, outcrossing risk was low, clonal propagation to produce stable transgenics was possible, tubers could be eaten, were used in food industry and tubers could be stored for long periods without refrigeration.

Tobacco and alfalfa have leaves which are major source of biomass; banana, tomato, apple, guava and strawberry were the fruit crops; peanut,corn, soybean and chickpea were seed-based crops; cabbage, lettuce, potato,carrot and spinach are the vegetable plants which were used for the production of vaccine antigens.Plant vaccines have been developed for many human diseases such as hepatitisB, Human Immunodeficiency virus , rabies etc. More than 25 vaccines licensed for use in humans with many more in the development pipeline . In March 2018, Medicago Inc. conducted phase III clinical trials to develop flu vaccine in tobacco and is expected to be launched in the market by 2020-2021. In June 2018, researchers from the University of Nottingham, Malaysia, launched a project to develop plant-based vaccine against dengue fever, caused by Aedesmosquitoes. Researchers from Arizona State University’s Biodesign Institute,developed a norovirus vaccine from tobacco plant in August 2018. Other than tobacco, many edible crop plants are also used such as the dengue virusvaccine produced in lettuce through chloroplast transformation in 2016.

MedicagoInc., iBio Inc., Icon Genetics-GmbH, Creative Biolabs etc. are involved in plant-based vaccine development. The Queensland University of Technology,Australia plans to use the genome sequence information of Nicotiana benthamiana to use the plant as biofactory to produce antibodies, vaccines and therapeutics,to develop protein-based diagnostic products in bulk quantities at a lowcost against COVID-19 or similar viruses or pathogens.Although plant-based human vaccines are not approved yet, vaccines against influenza, norovirus, hepatitis B virus, rabies virus etc. have been successfully produced in various transgenic plants and tested for their safety and efficacy under clinical trials . Vaccines against several animal diseases have been developed in plants, tested in animal models and in target animal species with the disease. Vaccines against anthrax, Bovine Herpes virus 1, enterotoxigenic E. coli etc. were produced in tobacco. The Foot and Mouth Disease virus VP1 epitope were expressed in alfalfa and Arabidopsis leaves and potato tubers, while the FMD virus polyproteinP1-2A/protease 3C was expressed in tomato. Epitopes of mink enteritis virus, murine hepatitis virus and rabbit haemorrhagic disease virus were produced through cowpea mosaic virus display in cowpea, Tobacco Mosaic Virusepitope display in tobacco and Plum pox virus epitope display in tobacco respectively.The infectious bronchitis virus S1 glycoprotein was expressed in potato tuber while transmissible gastroenteritis coronavirus glycoprotein N terminal domain was expressed in maize grains.

Bovine trypsin derived from maize has been commercialized since 2002.Neutralizing antibody responses were elicited against homologous and heterologous Newcastle Disease virus by inoculating plant-produced fusion protein antigen ,into Specific Pathogen Free chickens. Newcastle disease vaccine derived from tobacco cells was first approved for poultry use by United States Department of Agriculture .In addition to expression of antigen for vaccine production ,pharmaceuticals such as antibodies, enzymes, therapeutically important proteins or peptides and growth hormones are produced in plants . The vaccines used for immunisation against contagious disease are mostly costly and not easily accessible. On the contrary, the plant bioreactors are cost-effective and cheaper.

Red and red-far red combination resulted in the highest stem elongation but with stem fragility. When Lactucasativa of variety red curly lettuce was grown under different light spectrum, it was found that anthocyanin synthesis, protein content and phenylalanine ammonia-lyase enzyme activity were highest in combined radiation of blue and red-light treatment . In another study, where redand green basil microgreens were grown with blue and redLED, it was found that growth of microgreens was enhanced with predominantly blue illumination showing larger cotyledon area and higher fresh mass,enhanced chlorophyll a, and anthocyanin pigments contents. Stimulation of phenolic synthesis and free radical scavenging activity were improved by predominantlyred light in the green cultivar and blue light in the red cultivar,which indicates that LED light has an influence on the colour of the leaf.

Lettuce crops grown with red and blue LED lighting used50% less energy per unit dry biomass accumulated than under traditional light sources, which indicates that the significant reduction in energy consumption for plant-growth by using LED than traditional light sources . In an experiment on the indoor cultivation of basil and strawberry, it was found that the plants expressed increased biomass, fruit yield, antioxidant content and reduced nitrate content when treated with LED with highest energy use efficiency than traditional fluorescent lamps and spectral red: blue ratio of 0.7 was essential for proper plant growth with improved nutraceutical properties .From the literature review, it is found that when the microgreens are treated with red and blue LED combination, they showed highest photosynthetic activity but the exact ratio of blue and red-light combination at which the results are obtained are not evident.

Thus, it is essential to consider the production of microgreens at different ratios of red and blue combination under different light intensities. The environmental factors such as carbon dioxide, temperature and humidity play a crucial role in the growth and development of plants throughout their lifecycles. Their levels are specific and critical for different stages of plant growth. In an experiment conducted to determine the impact of free air carbon dioxide enrichment on development and progress in cotton , the results were found that carbon dioxide enrichment resulted in significant increase in photosynthesis and biomass of leaves, stems and roots, reduced evapotranspiration,changes in root morphology and increased soil respiration .When experiments have been conducted to determine the effects of atmospheric humidity on sugar beet, wheat and kale, the results were found that the plants showed increases in growth with increased atmospheric humidity.

The leaf area of sugar beet and kale plants was also increased with increased humidity due to the increase in number of cells . Each plant species will require optimum temperature to show maximum rate of plant growth and development. Both the atmospheric temperature and the soil temperature will influence different stages of plant development such as vegetative and reproductive stage. For instance, the vegetative growth of the plant requires comparatively higher optimum temperature than reproductive stage .There are many literatures pertaining to the effect of climatic factors such as carbon dioxide, temperature and humidity on plant growth and development both indoor and outdoor conditions but there are no such studies on microgreens. From the previous related works, it is important to consider their impact of climatic factors and their regulation to produce maximum growth and yield of microgreens which are grown indoor. Plants require both macro- and micro-nutrients for their growth and development.Vertical farming technology involves the soilless culturing of plants with application of nutrient solution, especially, inorganic fertilizer to the plants through soilless medium. The Hoagland solution developed by Hoagland and Arnon with definite composition is a hydroponic nutrient solution,which provides every essential nutrients for plant growth .

The composition of Hoagland solution is given in Table 2 .The commercial crops grown under soilless culture yield good quality produce when supplemented with the nutrient solution of specific composition based on the crop, the growth stage, the climatic conditions, the substrate, or hydroponic system used. Thus, the standard formula is not always applicable to all the crops . When experimenting the plants with continuous flow of nutrient solution, it was found that there is a certain minimum concentration below which uptake is no longer possible at the required rate. At the other extreme of concentration, luxury consumption may lead to internal toxicity . Thus, it is not necessary to apply nutrients unless required by the crop for its growth.

A total of one hundred and nineteen structured questionnaires were developed and thirty percentage of the root crop farmers,were randomly selected in each of the seven sampled community for interviewing.Questionnaires were distributed to educated farmers young and old, or who had educated relatives in their compounds after thorough explanations of the aim and objectives of the study to the respondents through an interpreter. The implication of including the younger farmers is that younger farmers are likely to adopt new innovation faster than the older ones. This is in agreement with that majority of farmers within the age range of 41 to 50 years are still in their active age, more receptive to innovation and could withstand the stress and strain involved in agricultural production and ease adaptation to climate change.Data analysis was carried out using simple proportion for qualitative variables while mean and standard deviation were used in analyzing the qualitative variable.

The outputs were further presented in tables and charts. The data was structured and presented from the responses to the research questions in line with the aim and objectives of the study. The analysis of the data both primary and secondary data was aimed at bringing to the knowledge of both informed and uninformed stakeholders, the basic understanding, adaptive strategies and experiences of climate change and its effects on root crop farming system in the sampled communities of the Ikwerre Local Government Area, of Rivers State,Nigeria. Analysis of how farmers adjusted to long-term shifts in two major climatic variables that affected root crop farming revealed that apart from Ozuaha all the other six communities indicated 100% wait for a favorable period before planting during a long-term shift in rainfall and replanting of crops that died off during long-term shifts in temperature. However, in Ozuaha, 3.4% will plant at their usual planting periods during long-term shifts in rainfall. But during long-term shifts in temperature, 96.6% of the farmer’s reported that they replanted those crops that died off and 3.4% do not replant.

Findings further showed that the four major root crops cultivated include sweet potatoes, cocoa yam, yams and cassava. Amongst these four major root crops, cassava is the most widely root crop cultivated, meaning that each sampled farmer cultivates cassava followed by yam, cocoayam and sweet potatoes.It therefore, showed that cassava was the mostly affected root crops by climate change especially the changes in rainfall and temperature. There is now unequivocal evidence that the climate of the Earth has demonstrably warmed up since the pre-industrial era and that most of the warming over the last 50 years is very likely to have been as a result of increase in Greenhouse Gases concentrations in the atmosphere .Three major physical impacts of climate change, according to IPCC in Ghana, are temperature rise, changing rainfall regime towards a longer dry season and a varnishing wet season. also underscored the fact that there has been a noticeable shift in the rainfall pattern in Ghana towards a longer dry season and the disappearance of short dry spells between major and minor seasons.

Agriculture is the pillar of Ghana’s economy providing employment to the masses in the rural areas and sustaining the vast majority of the population. Only about 0.89% of the country’s arable land, equivalent to23,657 hectares is cultivated under irrigation, benefitting only rice production .According to , maize yields in Ghana are low compared to that of developed countries. The national average yield is about 1.6 mt per hectare as compared to a country like Brazil . This is primarily due to the inadequate input of fertilisers and non-existing irrigation facilities in the maize production areas .Among the staples cultivated in the country, maize has been identified as one of the most essential food crops within the grains . Maize is grown on more than 997,661 hectares across all the various agro-ecological regions in Ghana .Generally, high temperatures have been reported as detrimental to grain yields. It was projected that maize yields would drop from its estimated level 0.5 to 6.9 in the year 2020. The study of indicated that each day’s temperature above 30˚C would reduce the final yield by 1% under optimal rain-fed conditions, and by 1.7% under drought conditions. In the transition and forest belts, though droughts are not the main drawback, variations in rainfall deficits, particularly in the number of rainy days pose new challenges to rural livelihoods . There is a decrease in the farming season in several places as well as a gradual waning of the secondary growing season in the transition zone . Both primary and secondary data were used in obtaining the data.

With regards to primary data, two focus group discussions were organized in two communities within the Municipality. Twelve key informants comprising seven maize farmers, two sub-chiefs, one meteorologist from GMet-Wenchi branch, one agricultural extension officer from Nkoranza SouthMunicipal Assembly and the 2015 best farmer of the Municipality were also interviewed.Secondary data consisted of meteorological data from GMet which covered 1960 to 2014. This data was divided into two tranches, 1960 to 1982 and 1988 to 2014.

Among the precipitations, the Northeast precipitation is caused by occluding cyclones above the Sea of Japan, while the East border precipitation is induced by the same cyclone but formed at sea level. The Southwest precipitation is wrought by the frontal effects between southwest moist wind and the northwest dry wind. The West to east precipitation is due to the change of pressure pattern inside the Region, and the Inland precipitation is induced by ground breezes.Comparing to other typical precipitations through the 30 winters, the Coastal precipitation in January 2017 is not seen, while the West to east precipitation and Inland precipitation are additional types in January 2018. The former can also be contributed to existence of cyclones above the ocean surface because ofits location, and the latter is mainly due to the pressure change and local breezes of the Region.

The high amount of monthly total precipitation in January 2018resulted from a relatively high precipitation frequency, rather than an augmentation of precipitation intensities. Thus, the high precipitation amount in January 2018 is caused by active atmospheric circulation above the Region. For the Region in January 2018, the maximum of the monthly total precipitation is 4 times more than that in other winter months for the recent 30 years. The highest value of precipitation mainly concentrates along the mountainous areas.For these precipitations, there are five major types of precipitations named after their locations: Northeast precipitation, Southwest precipitation, West to east precipitation, East border precipitation and Inland precipitation. It is found that a continuous moderate precipitation phenomenon is dominant in the Region in January 2018, instead of increase in either precipitation intensities or number of intense precipitations. Such precipitations are mainly caused by cyclones in the vicinity of the studied Region.

The Northeast precipitation and the East border precipitation are both induced by cyclones above the Sea of Japan, but the former is caused by an occluding cyclone while the latter by a small cyclone formed at sea level. The Southwest precipitation is frontal precipitation between southwest moist wind and northwest dry wind. The West to east precipitation is generated by the change of atmospheric pressure above the Region, and the Inland precipitation is risen by ground breezes.For future work, the causes of such cyclone frequent weather in the Region should be exploited from a global perspective. More work should be investigated in the relationship between the cyclone occurrence frequency in the Region and the Indian Ocean Dipole, the El Nino Southern Oscillation and the Pacific Decadal Oscillation. Climate change and other global drivers of socio-economic, energy, global trade, resources and demographic changes are set to affect present and future human development including the vulnerable areas such as southern Africa region.

Faced with imperatives of increased food production and poverty alleviation, present day conditions call for high vigilance in developing and preserving the raw materials for food production, specifically through land and water management. Faced with imperatives of increased food production and poverty alleviation, present day conditions call for high vigilance in developing and preserving the raw materials for food production: land and water. Agricultural water holds significant scope for addressing climate change vulnerability and adaptation needs as well as water and food insecurity in the region.The observed human-induced changes to climate pose a threat to food security the world over and Southern Africa is no exception. Climate model studies show that the average temperature of earth’s surface is expected to increase by 3˚C over the next century, if greenhouse gas emissions continue to rise at the current rates. This has a negative impact on crop yields and food security in the Southern African region where 60% -80% of the population is directly dependent on agriculture for their livelihoods .Crop and livestock production systems will have to change in response to the changing agro-ecological conditions.This manuscript aims at developing a set of practical approaches to agriculture in order for farmers to beresilient and adapt to the predicted climate changes.

Here, a regional framework for the implementation of climatesmart agriculture concepts is also illustrated.Also it will support investment decisions in climate change adaptation. This paper seeks to inform scientists and experts in the fields of agriculture, climate change and socio-economics to collectively build a strong base of evidence on climate change and variability impacts on rain fed cropping systems. It also tries to devise agricultural interventions and practices that enhance general resilience in the quest to overcome climatic shocks and develop adaptation strategies.

The latest record from government agency shows that there is an estimate of 6.8hectares of forestlands with a net loss of 400,000 hectares every year.Therefore, to maintain sustainable food production while providing housing and industrial uses to lands, House Bill 4750 or the “Urban Agriculture Act of 2011”, and House Bill No. 720 known as the “Urban Agriculture Act of 2013”, mandate the Department of Agriculture and Department of Science and Technology, to promote the use of urban agriculture and vertical farming. These two laws aim to ensurefood security and rejuvenate the ecosystem functions. The law further mandates that abandoned government lots and buildings owned by the national or local governments or available land resources of state universities and colleges shall be considered for growing crops, raising lives tocks purposely to sustain the demand for food production using vertical farming and urban gardening.

Furthermore,the methods would be included in the academic curricula in secondary and tertiary levels of education both in the private and public schools . The Philippines is an agricultural country with a land area of 30 million hectares.47 per cent of which is agricultural land. Of the total area used for food crops,Two Hundred Seventy Thousand hectares are used to grow vegetables and root crops. Among them is Bulb onion . It is a favorite culinary in gredient in the Philippines and in Southeast Asia. There are about 12,000 hectares of farm lands planted with onion in Nueva Ecija and Ilocos region. The province of Nueva Ecija is the leading producer of onion accounting for 99 percent of the region’s onion production. The weather condition in the province experiences two seasons a year. The dry season during the months of November to April while the wet season occurs during the rest of the months of the year.Among the municipalities in Nueva Ecija, Bongabon is known as the onion capital of the Philippines because of the volume it produced.In this study,shallot variety of onion is used because it can be grown almost throughout the year.

At present, onion production in Nueva Ecija is threatened by importation,smuggling, and the emergence of pests bringing havoc to potential earnings of onion farmers. There are about 3000 hectares of onion plants affected by “armyworms”resulting in the dramatic decline in the production of this crop in theregion. There are three reasons cited for the decrease in onion production in Nueva Ecija: 1) The prolonged rainy season; 2) Farmers losing their marketsh are because of the coming in of imported onion; 3) Emergence of insects causing the drop in harvest.At this juncture, the literature shows that development of the onion industry in the Philippines is facing a challenge. The declining production of the industry demands that new model and a new system of farming needs to be reconsidered.Despite the present state of the industry, there is no sufficient number of researchers applying new technology and method of making onion production viable.A necessary means to maintain its mark as the onion capital of Southeast Asia. Therefore, the present study aims at using the technology of vertical farming and hydroponics as a viable system of producing onion. Investigate on the possibility of adopting vertical farming technology which might serve as a potential solution to the problems facing onion production in Nueva Ecija.

In view of the above discussion, the study posed the following statements of the problem: 1) How may the need for a sustainable system of production of onion be described in terms of viability; 2) What is vertical farming using hydroponics;3) How may vertical farming using hydroponic system be described in terms of, production cost; usability for onion production; manageability of farm activities and acceptability to farmers and onion growers? 4) How may the vertical farming using hydroponics be described in Nueva Ecija; 5) How may the findings of the study contribute to agricultural productivity in the onion industry in Nueva Ecija? Vertical farming or high-rise farming is a proposed indoor, urban farming technology involving large-scale agricultural production in multi-story buildings. While, hydroponics and aeroponics methods are tools to produce crops insoil-less nutrient solutions.Vertical Farming promises to eliminate external natural processes since crops will be grown under carefully selected and well-monitored conditions, ensuring an optimal growth rate for each species of plant and animal year-round . Iteliminates the need for fossil fuels to run machinery during the different stages of farming .It is estimated that one acre of vertical farming is equivalent to ten to twenty soil based farmlands .Vertical Farming has the following advantages compared to traditional farming practices: 1) Year-round crop production as it requires no soil rejuvenation and soil mineral recovery ordinarily observed in land-based farming. 2) Significantly reduces farm inputs and the use of fossil fuels causing damage to the environment.Land-based agriculture requires land plowing using farm implements burning gasoline and other petroleum products. 3) The technology can be put up anywhere; 4) it minimizes weather and crop failures as it is under a controlled environment the farmer himself sets; 5) it helps in restoring the farmland to the ecosystem. Hydroponics is one of the many methods of soilless agriculture through which plants are grown in mineral nutrient water .

Specifically, water stress resulted in the lowest increase in leaf area development , and leaf size tended to decrease with decreasing water supply. A study found that leaf area was strongly negatively correlated with leaf water loss,suggesting that bigger leaves of P. sibirica and U. pumila in 4 and 8 L h−1 irrigation levels may be less prone to water loss via evapotranspiration. The efficient use of irrigation and fertilizer has recently gained much attention in reforestation efforts in arid regions because of the highly variable or limited rainfall events. In this study, we revealed that the interaction of 4 and 8 L h−1 with NPK or COMP positively increased LB of all species. This can be attributed to enhanced cell divisionand physiological activities under water-sufficient or high turgor pressure conditions and improved nutrient availability.

Our result is consistent with that of some water and fertilizer experiments conducted in drylands . Similarly, Zhang and Xi observed that the aboveground plant biomass increased when precipitation increased and decreased when precipitation fluctuated significantly. A significant increase in aboveground biomass and leaf area was also detected when the plants were supplied with nitrate, and such a result was associated with the effects of nitrate on cell turgor pressure . Thus, the current result suggests that sustainable reforestation in arid and semiarid regions in Mongolia can be achieved with the use of a combination of the appropriate amount of water and fertilizer. This is because excess water and application of both organic and inorganic fertilizers can exacerbate land degradation and environmental damage in arid and semiarid regions. Contrary to our expectation, CONT and a low amount of irrigation have generally shown to have a higher total chlorophyll content compared with a high amount of irrigation with/without NPK or COMP, particularly in 2019 for both species.

The result of the present study agrees with the findings of Hassanzadeh et al. , who reported that irrigating the plots under flooding conditions led to a decrease in chlorophyll content and senescence, whereas drought stress increased chlorophyll content. Our result is attributable to the effects of moisture on photochemical activity and chlorophyll synthesis in leaves. Conditions such as excess water and nutrient availability may have hindered the chlorophyll synthesis of irrigated plants, particularly those grown in 4 or 8 L h−1 with NPK and compost. Several studies explained that low levels of chlorophyll in leaves might be poorly related to water conditions in the field because of interacting effects of the other environmental factors, such as soil characteristics, light, and air temperature . A change in the amount of far-red radiation and light reflectance of leaves was also cited as one of the reasons for the change in chlorophyll content with increasing soil moisture .

A study by Hamblin et al. also mentioned that the reduction in leaf chlorophyll content can reduce the heat load, thereby reducing water requirement to cool leaves. Nitrogen is an essential part of the chlorophyll molecule; hence, the lack of it in the mineral nutrient supply may significantly affect chlorophyll synthesis in plants. Here we revealed that adding NPK or compost to a higher amount of irrigation significantly decreased the chlorophyll content compared with CONT in all species, implying that some factors such as mineralization, leaching, and volatilization may have come into the picture. Because of the high amount of irrigation, the applied fertilizer may have been leached out, leading to N deficiency in plants. Another possible reason is that the high amount of irrigation may have facilitated the mineralization of the applied fertilizer in the soil, leading to a luxury consumption of N by plants. Such a luxury consumption produced excess N that may not be metabolized into functional or structural compounds necessary for chlorophyll synthesis. One study, however, reported that the stress-induced loss of chlorophyll was not linked to a lack of nitrogen .

This is probably because of the effect of water stress on nitrogen mineralization and uptake. Further, the significant reduction in leaf chlorophyll content may be a consequence of increasing the LA, SLA, and LB of the plants grown in a higher amount of irrigation with NPK or compost. The scientific name of the oriental persimmon is Diospyros kaki, and it is classified as a deciduous tree belonging to the family Ebenaceae of the genus Diospyros. The persimmon originates from northern China and is said to have been introduced to Japan during the Nara period, which took place over the years 710–794. The astringency of the peels of persimmon is caused by tannin, which is a condensation product of epigallocatechin,epicatechin, and their galloyl compounds. When persimmons ripen, acetaldehyde is generated within the flesh of the fruit and forms cross-links with the water-soluble tannin to form polymers, making the tannin insoluble and thus reducing the persimmon’s astringency .

Australia is the largest producer, with 47.1% of the global production, while Europe is second according to FAOSTAT . In Europe L. angustifolius L. and L. luteus L. are the predominant cultivated species in the north and countries with more than 10,000 ha of lupins are Poland, the Russian Federation, Germany, Belarus, and Ukraine. In the south, where L. albus L. is predominant, Italy , France , and Spain are the main lupin-producing countries . Modern breeding efforts to improve white lupin agronomic characteristics are very recent . Thus, there is still significant variability even among commercial germplasm for the most essential breeding targets, namely, the seed’s nutritional value and the toxic, bitter secondary metabolites QAs . The total alkaloid content in white lupin varies from 0.02 to 12.73% of the seed’s dry weight. Cultivars possessing the pauper gene contain 0.02–0.05% alkaloids of the seed dry weight . The recently published white lupin pangenome study demonstrated that pauper locus has a key role in the species domestication and breeding .

Other important agronomic characters of white lupin, which attracted the breeders’ attention, are vernalization insensitivity , anthracnose resistance , yield stability , and abiotic-stress acclimation . White lupin’s global commercial potential has incited the breeding interest to focus not only in yield boost, but also in expanding its cultivation to agroclimatic regions, other than the Mediterranean basin, as extreme climate-change-related phenomena push the cultivation of some crops northwards . On top of that, restricted precipitation levels during spring and frequent dry spells, throughout the Mediterranean basin, as a consequence of climate change , have a detrimental impact on pollen fertility , pollinator-flower interactions , pod filling, and seed development, resulting in premature harvesting and yield losses.Anthracnose is a global fungal disease, responsible for devastating epidemics, characterized by signifificant yield losses . Temperatures over 10 ◦C and humid weather promote conidia germination, with 25 ◦C being optimal for fungal growth; whereas dry summer conditions are favorable for the preservation of inoculum on unharvested plant tissues .

Colletotrichum lupini, is mostly identified as the responsible pathogen for lupin anthracnose. Nonetheless, it has been reported in several other crops such as olive . The pathogen emerges as an alarming polyphagous phytopathogenic strain for the Mediterranean agriculture. White lupin breeding has been directed to the creation of elite anthracnose resistant cultivars, employing map construction, genomic screening, phenotyping tools, fifield experimentation, and generation of molecular markers, to detect anthracnose resistant accessions . Thus far, Ethiopian landraces have been extensively studied, revealing a highly diverse germplasm and embodying unique loci that confer resistance to anthracnose . Successful breeding significantly depends on the extent of the available genetic resourcesPhenotypic and molecular markers have already been used in few studies, to estimate the genetic diversity between wild and breeding white lupin germplasm, and to enable incorporation of potentially valuable alleles from distantly related wild accessions tothe genetic pool of elite cultivars .

Additionally, molecular markers are continuously developed, for the effective selection of germplasm with desired traits . Furthermore, genomic resources are now available in white lupin and their use will greatly advance our understanding of the species diversity. In such an effort, very recently genome sequences of 39 accessions were used to establish a white lupin pangenome that can be used as resource to identify genes linked to important agronomic traits and analyze genetic variability. Although such progress will inevitably lead to development of more sophisticated tools to explore genetic variation in white lupin genetic resources, up to now SSR markers have been proved integral tools to investigate species diversity. The Balkan Peninsula represents a yet untapped germplasm diversity center for white lupin, concealing potentially valuable loci in landraces and natural populations that could promote adaptability to climate-change-relevant extreme conditions . In this study we applied available SSR molecular markers linked to agronomically important traits and morphological seed characteristics to explore the genetic diversity of white lupin Greek landraces and compare it with that observed in commercial varieties and breeding lines.

Results of the study may facilitate marker assisted breeding in white lupin and enable identification and introgression of valuable alleles into new elite cultivars. All seed morphological characters’ measurements were statistically analyzed and a summary of the statistics is shown in Table 1. The Multivariate Analysis of Variance indicated that all 45 accessions are distinguished from one another, when regarding their seed morphology, with statistical significance . A Pearson’s correlation matrix revealed strong correlation between seed area with TSW, perimeter and width and between mean Gray Value with maximum and median Gray Value , suggesting, that a single character measurement is sufficient to represent highly correlated characters.

We previously showed that this compound has a remarkable antibacterial activity at low concentrations: 0.24 µg/mL against Staphylococcus aureus and 3.9 µg/mL against Escherichia coli . As such, we have strong reasons to consider that BrCl-flav has a good potential for the development of new antimicrobial agents. Therefore, further studies using four clinical Candida isolates were employed to investigate BrCl-flav antifungal properties and mechanism of action. A potent antifungal effect was evidenced for BrCl-flav against all Candida strains tested in vitro using the determination of minimum inhibitory concentration. The method allowed us to assess the lowest concentration of BrCl-flflav which inhibited the growth of the tested fungal strains—15.62 µg/mL.

When compared with the reference drug fluconazole, a considerable higher antifungal activity was recorded against fluconazole resistant C. albicans, C. krusei and C. glabrata isolates. Compared to most of the reported natural and synthetic flflavonoids, our compound displayed a stronger antifungal activity, being up to 28-fold more active against Candida spp.. We must emphasize that BrCl-flav exhibited an antifungal activity comparable to some chalcone, flavones and flavanones derivatives considered to be the most potent synthetic flavonoids against Candida reported so far. The fungistatic effect was dose-dependent, increasing concentrations of BrCl-flflav progressively inhibited the fungal growth of all tested Candida isolates.

The cells incubated with 7.81 µg/mL BrCl-flflav showed no significant growth inhibition compared with control. On the other hand, a signifificant growth delay represented by prolonged lag phases occurred when Candida spp. cells were incubated in the presence of BrCl-flav at 15.2 µg/mL, corresponding to MIC. We must point out that no turbidity was recorded by spectrophotometric measurements for all Candida cells exposed to 31.25 µg/mL within the time span of the experiments , denoting a strong fungicidal activity. A time-kill kinetics assay was employed to determine the fungistatic or fungicidal activity of BrCl-flav over time. The analysis confirmed also the important antifungal activity of BrCl-flav against all Candida spp. after exposure to concentrations equivalent to MIC and 2 × MIC . One exception occurred for C. albicans strain—a fungistatic effect was recorded by the growth experiments up to 24 h, while time kill studies revealed a fungicidal effect at 24 h.

We must emphasize that no viable cells were detected starting with 12 h after BrCl-flav exposure at 31.25 µg/mL , suggesting an important fungicidal potential. The fact that this activity has been recorded against fluconazole-resistant Candida strains makes BrCl-flav a more interesting compound for practical applications. Our conclusion is supported by a comparative literature survey which revealed that BrCl-flflav has higher fungicidal activity compared to many natural flavonoids.Also, the recorded activity was higher or comparablewith other synthetic flavonoids such as different chalcone, 1,3-thiazole and 2-hydrazinyl- 1,3-thiazole derivatives.We must emphasize that BrCl-flav acted as a more potent fungicidal compared with fluconazole, considered to be a gold standard of antifungal agents . Sorbitol binding affinity assay was used to investigate the interference of BrCl-flav with the Candida cell wall. Sorbitol is an osmotic protector which can support cell growth when the fungal wall is targeted by antifungal agents. In the absence of sorbitol, the fungal growth is inhibited due to the disruption of the cell wall. The effect is detected by an increase of the MIC value in the presence of sorbitol compared to the MIC value determined in medium without sorbitol.In our study, MIC values of BrCl-flav did not change in the presence of sorbitol, suggesting that the tested synthetic flavonoid does not target the cell wall. Therefore, other potential cellular targets were investigated. Penetration of PI into dead or injured C. albicans cells was evidenced using fluorescence microscopy. PI is a cell membrane-selective permeable dye that can only pass through damaged or permeabilized cell membranes, binding to DNA and exhibiting characteristic red fluorescence.

Our results revealed a gradual increase of the fluorescent cells number with the increasing concentration of the tested antifungal. After 24 h all cells exposed to BrCl-flav at a concentration equivalent to 5 × MIC were fluorescent, suggesting that the tested antifungal significantly damaged the fungal cell membrane integrity. Usually, damages to the cell membrane are related to cell lysis. SEM image analysis showed severe alteration of the cell morphology, with collapsed cells, wrinkled surfaces, along with cellular debris resulting from the disintegration of the BrCl-flav treated cells. These morphological changes are most likely caused by cell lysis.

The global recognition of the importance of this approach in also reflected in the G20 Rome Declaration from the recent Global Health Summit, held in May 2021 . The locution of One Health is undoubtedly not novel to parasitologists, who, by vocation, study infections and their causative agents and hosts, at the interface with human and veterinary medicine. With special regards to parasitic conditions, Africa harbours a plethora of them that require to be addressed according to the One Health approach, due to the dependence of parasites and their potential vectors or intermediate hosts on numerous environmental factors , habitat conditions such as temperature, humidity, availability and type of waters , etc.. For instance, malaria, the parasitic disease par excellence of the African continent, provides a compelling example of a condition that needs to be tackled through One Health efforts, due to the zoophilic behaviour of several competent mosquito vectors .

This is the approach taken by recent investigations aiming at treating cattle and other livestock with ivermectin or macrocyclic lactone-based “endectocides”4 in order to reduce the fitness of competent malaria vectors feeding on them . Similarly, the application of pyrethroid-based insecticides or arthropocides to cattle not only aims to control livestock-specific ectoparasites but also to reduce populations of zoophilic mosquitoes responsible for malaria transmission . These methods could be considered as complementary tools that could help reduce malaria incidence in a given area, although alone they may not suffice to eradicate the disease. It is nonetheless essential that control campaigns consider all hosts on which competent mosquitoes feed and the outdoor component of malaria transmission. Other major parasitoses of the African continent are ascribed in the group of the so-called Neglected Tropical Diseases , of which Africa bears ~40% of the world’s burden . These are typical diseases of poverty, highly endemic in rural areas, not coincidentally where hygiene conditions and water sanitation are poor and where tight cohabitation between humans and animals occur .

Out of the 20 NTDs recognised by the WHO, 19 occur in Africa, with 11 of them being of parasitological aetiology. Of these, at least seven include zoonotic agents . Although not involving a parasitic causative agent, NTDs like dengue and chikungunya still require parasitic arthropods as vectors , further highlighting the relevance of parasitological and entomological expertise in the study, management and control of most NTDs in Africa and elsewhere .Undoubtedly noteworthy in the area of One Health are also the so-called Neglected Zoonotic Diseases , that are deemed as “neglected” due to under-reporting on their occurrence, leading to an underestimation of their relevance to policy-makers and donors . The NZDs comprise a subgroup of NTDs , all of which except rabies are of parasitic aetiology. Currently, rabies, echinococcosis, foodborne trematodiases and taeniasis/cysticercosis are considered as priority NZDs by the WHO Department of Control of NTDs ; the fact that three of them are parasitic infections further highlights the topicality and heavy burden of parasitesin the realm of zoonoses. In addition, zoonotic causes of non-malarial febrile illnesses such as anthrax, brucellosis and leptospirosis are also recognised by the WHO to be of growing importance and are therefore monitored accordingly . The control of NTDs is a fifirm component of the UN 2030 Agenda for Sustainable Development, as reflflected in its Sustainable Development Goal target 3.3 aiming, by 2030, to “end the epidemics of AIDS, tuberculosis, malaria and NTDs ” .

In the past decade, some substantial progress was made in these regards, with 31 countries having succeeded in eliminating at least one NTD, eight of them being in Africa. However, despite these results, many of the targets set by the WHO in the 2020 roadmap were not met, in Africa and other endemic regions . In particular, the overall control of NTDs was negatively affected by the disruptions caused by the COVID-19 pandemic in the year 2020, with major hindrances being recorded in the delivery of health services . Accordingly, a new NTD roadmap was defined for the period 2021–2030, identifying critical gaps and actions required in order to prevent, control, eliminate or eradicate the 20 target diseases and disease groups . Importantly, the new agenda fully acknowledges the value of adopting an integrated One Health strategy for NTDs, promoting stronger multisectoral collaborations among agriculture, livestock, wildlife, environment, food safety, health and other ministries . Such a strategic development derives from the consideration that all zoonotic NTDs, in order to be fully controlled in human populations, need to be managed appropriately and possibly synergistically in animal hosts and/or reservoirs. Notably, tackling NTDs involving livestock species in their epidemiology , can provide dual benefifits in terms of human and economic development, by reducing risks of zoonotic infections, on one hand, and improving communities’ livelihood, through better animal health and productivity, on the other .

Previous documents show that increased photosynthetic capacity due to Mo supply contributes to the accumulation of carbohydrates, such as fructose and glucose.We observed that lipids, fibers, and saponins were significantly increased in CA1. Flavonoids were increased reasonably in CA3.Different impacts of Mo were observed in different species/cultivars, which might be due to different genetic backgrounds and the variable uptake of Mo.Phenolics play essential roles in plant development; these aromatic benzene ring compounds with one or more hydroxyl groups are produced by plants mainly for protection against stress .

In our study, the Mo priming of Canavalia seeds significantly increased the phenolic content of CA3 sprouts; a non-significant increase was found in CA2 and, surprisingly, a slight decrease was noticed in CA1. This might be due to differences in endogenous molecular pathways of different species. This link to the inherent levels of mechanisms may also lead to differences in the enhancing patterns of the individual phenolic compounds after Mo treatment, which we observed in our study. A similar trend was observed in DAHPS, the first enzyme of the shikimate pathway, which converts PEP and E4-P into 3-dehydroquaianate. Further, to assess the impact of Mo seed priming and enhanced phenolic production on the pharmacological properties of plants, we performed antioxidant and antidiabetic assays. FRAP, ABTS, and DPPH assays were used to evaluate the antioxidant capacity of species samples spectrophotometrically.

An increase in antioxidant potential was observed after Mo treatment in all species. In the FRAP assay, the increase was significant in both CA1 and CA3. CA3 also expressed a significant changein antioxidant activity, measured via ABTS assay. This correlates to the above findings that more impacts of Mo on synthetic pathways were observed in CA3. Antioxidant activity has recently become a target for product development in the pharmaceutical and cosmetics industries .Canavalia species are of medicinal importance due to their potential antioxidant properties. In a study, Canavalia gladiata extract, at the concentration of 2 mg/mL, showed an antioxidant effect comparable to that of ascorbic acid of the control group . Plant growth and antioxidant capacity are greatly dependent upon N availability. Higher N improves the stress tolerance of plants via enhancement of the antioxidant ability and inhibition of lipid peroxidation. Mo primarily improves the nitrogen fixation to the plant and increases its antioxidant potential, which we observed in our study. This also might be the reason for the enhanced antidiabetic potential of Canavalia species assessed by GI, α-amylase inhibition activity, and α-glucosidase inhibition activity.

Terpenoids and flavonoids of Canavalia gladiata are reported to play a role in lowering glucose levels and possessing antioxidant potential.The Mo-mediated enhanced concentrations of terpenoids and flavonoids might have led to the increased antioxidant and antidiabetic activities of Canavalia sprouts in our study. For sample preparation, we used an ETA 0067 grinder with grinding stones, VIPO mini grinder, followed by homogenization by Vibrom S2 , and a cryogenic grinder accompanied by liquid nitrogen. Supercritical fluid extraction using SE-1 extractor and a steam distillatory apparatus according to CSN 58 0110 and CSN 6571 were successively applied for extraction and subsequent determination of the total content of Canavalia oils. Approximately 500 mg of each sample was transferred into an extraction column for SFE extraction. The extraction was performed at 40 MPa for 60 min, and extractor and restrictor temperatures were 80 and 120 ◦C, respectively. The extract was further trapped into a hexane layer inside a trapping vessel.The determination of phenolics and their precursor metabolites was carried out using an ultra-performance liquid chromatography system coupled with a quadrupole mass spectrometer provided with an ESI source according to Wang et al.’s method.

Phenolic and flavonoid levels were identified by comparing the standard mixture of different phenols and flavonoids to the relative retention time. The concentration of each compound was calculated using the peak area of the corresponding standard. In addition, deoxy-d-arabino 201 heptulosonate-7-phosphate synthase activity was analyzed according to Wang et al., . This enzyme catalyzes the reactions in cinnamic and shikimic acid pathways that are involved in phenylpropanoid biogenesis and therefore in the biosynthesis of coumarins. Samples were first homogenized in 3 mL 50 mM Tris-HCL buffer . The assay mixture contained 0.1 mM erythrose-4-phosphate, 0.2 mM phosphoenolpyruvate, and 0.1 mM MnSO4/0.1 mM CoCl2. In total, the reaction was initiated by enzyme addition and terminated by 25% trichloroacetic acid addition.