As such, more work can be done on how disturbances alter seed microbiome assembly processes and outcomes.The genus Vitivirus was created in 1997 for the classification of type member grapevine virus A , a plant virus discovered in grapevine with a filamentous flexuous particle differing from trichoviruses in its genomic arrangement. Vitiviruses have a single-stranded RNA genome encoding five genes: replicase , movement protein, coat protein , nucleic-acid-binding protein and a 20 kDa protein of unknown function. In the 2018 International Committee of Virus Taxonomy Master Species List , nine species of vitivirus infecting grapevine are recognized: Grapevine virus A, Grapevine virus B, Grapevine virus D, Grapevine virus E,Grapevine virus F, Grapevine virus G, Grapevine virus H, Grapevine virus I and Grapevine virus J. Since 2019, two more proposed vitiviruses were discovered in grapevine. Grapevine virus L was initially identified in RNAseq data and later detected in multiple plants in Croatia, New Zealand and the United States. Grapevine virus M was also discovered by high throughput sequencing in an American hybrid grapevine. Three different vitiviruses have been associated with the etiology of rugose wood disease in grapevine, a disease with world-wide distribution.
GVA is associated with stem grooving on the variety Kober 5BB, grapevine virus B was identified as the causal agent of corky bark in the variety LN33, procona florida container and grapevine virus D was implicated in growth reduction in the rootstock Freedom. Additionally, these vitiviruses are frequently detected in coinfection with grapevine leafroll viruses, resulting in synergistic interactions that can lead to lethal effects in several scion and rootstock combinations. The potential pathogenic role of the remaining grapevine vitiviruses, including proposed members, is still unknown. Reliable diagnostic methods are critical in determining the viral infection status of a grapevine. Multiple tests are available for the detection of vitiviruses, including biological indexing, real-time or end-point reverse transcription PCR and HTS. Biological indicators do not show symptoms for all vitiviruses infecting grape, and RT-PCR assays can fail to detect vitivirus variants containing nucleotide differences at critical primer binding locations. HTS is the most effective means of detecting all vitiviruses but its high cost at large scale limits its use as a screening tool. HTS data is helpful to inform and update RT-PCR primer design as new virus strains are continually being characterized. In this study, a universal end-point RT-PCR assay involving degenerate primers with the capacity of detecting all the known grapevine vitiviruses was developed.
To validate the new assay, eleven grapevines each infected with one of the vitiviruses were tested. Moreover, a field survey was conducted of known vitivirus-infected grapevines. Following the first reports of vitiviruses in grapevine, several vitiviruses have been discovered in other hosts; consequently, we investigated if the universal assay can detect these vitiviruses.A universal assay able to detect all known grapevine vitiviruses and potentially other members of the genus Vitivirus was developed here based on sequence data available in GenBank. The presence of highly conserved motifs in the REP protein allowed the design of end-point RT-PCR primers, providing an alternative assay to reduce the work associated with the diagnosis of vitiviruses. The extensive sequence divergence existing among grapevine vitiviruses, observed at the nucleotide and aa levels, makes it difficult to design a test with broad-range detection. RT-PCR with degenerate primers is a simple strategy that is frequently used for the specific and simultaneous detection of multiple viruses. Assays involving degenerate primers targeting grapevine vitiviruses have been described before, however, these studies were conducted in the pre-HTS era, when fewer vitiviruses were known and sequence data was limited.
Although Vitis spp. is recognized as the main host associated with the genus Vitivirus, vitiviruses have been identified in other perennial hosts, the majority of which are woody plants. For example, vitiviruses have been reported in blackberry, mint, agave and recently in blueberry. The universal assay successfully detected MV-2 in mint, however failed to detect BGMaV in blueberry. Additional investigation revealed a variation in motif A of BGMaV , and a similar scenario was observed in AVV. Based on our PCR results we predict that the universal assay will miss AVV during diagnosis, though, the rest of the known vitiviruses do not display any a discrepancy in motifs A or B and they should be detected by the assay. The family Betaflexiviridae comprises twelve different genera , including Vitivirus, Trichovirus and Foveavirus. After in-silico and in-vitro analyses of trichoviruses and foveaviruses, we did not find evidence for cross-reaction by the universal assay. A single test for all known grapevine vitiviruses can be a useful tool for improving efficiency and reducing costs of large-scale surveys. Potentially, this generic assay may detect novel Vitivirus species in grapevine and other hosts given its unbiased nature. Similar assays have been developed for carlaviruses, nepoviruses and different members of the family Betaflexiviridae. Grapevine is clonally propagated, consequently, to prevent the spread of vitiviruses, it is critical to use virus tested material. The assay developed here will be made available to diagnostic labs and will facilitate the production of certified virus-tested propagation material and the effective control of vitiviruses.Over the last few decades, the field of nutrition has grown and evolved. Although we continue to define the critical roles that nutrients play as fuel sources, enzyme cofactors, signaling molecules, and vital infrastructure for our bodies, the cutting edge of nutrition research is pushing beyond simply meeting our bodies’ basic needs. Indeed, as the population is living longer, an emerging focus for nutrition has been on obtaining and maintaining optimal health over the life course. On 10 October, 2022, the Council for Responsible Nutrition held their annual Science in Session conference entitled Optimizing Health through Nutrition – Opportunities and Challenges. The audience consisted of scientists and executives from dietary supplement and functional food companies as well as nutrition graduate student awardees of a CRN and ASN Foundation educational scholarship to attend the symposium. CRN is a trade association representing dietary supplement and functional food companies. The goals for this meeting were to propose a definition for optimal nutrition and identify strategies and tools for evaluating optimal health and nutrition outcomes while highlighting the gaps in this emerging space. Now more than ever in history, our population’s health has emerged as a global priority. Currently, 6 in 10 adults in the United States have a chronic disease, and 4 in 10 have 2 or more. In <10 y, the number of older adults is projected to increase by ~18 million. This means that by 2030, 1 in 5 Americans is projected to be 65 y old. As the major risk factor for many chronic illnesses is age, it is anticipated that the rates of all age-related diseases, especially chronic diseases, will skyrocket, potentially overwhelming the health care system. We need to enable the health care system—and the population—to be more proactive rather than reactive toward health outcomes. There is a critical need to help find solutions to optimize health across the lifespan to support living better longer, i.e., health span. Ensuring optimal nutrition is a significant and easily modifiable variable in the solution for maintaining and improving health span. We need to advance concepts beyond essential health and consider meeting the nutritional needs for optimal health. Although the nutrition science community is moving toward the vision of nutrition to support optimal health, many challenges and gaps still exist, but there are also recent advances and exciting opportunities.
The goal of the CRN “Science in Session” workshop was to discuss these challenges, gaps, and opportunities in order to advance the concept of nutrition for optimal health. This review summaries these findings and discussions.The DRIs for individual nutrients, procona London container including the Estimated Average Requirement and the RDA, are life stage- and sex specific recommendations for Americans and Canadians. These reference intakes were established in the 1990s by the Food and Nutrition Board of the National Academies of Sciences, Engineering, and Medicine to prevent deficiency disease and to reduce the risk of chronic diseases. However, incorporating chronic disease endpoints has been extremely challenging, primarily because data are largely lacking. Thus, the current DRIs, including the RDAs that are aimed to cover the nutrient needs of 98% of the population, do not account for the amount of a nutrient that one needs in order to achieve and maintain ‘optimal’ health.The science of resilience is not a new concept—this scientific concept was documented in the literature as early as the 1800s; the terminology entered the biomedical sciences in the mid- 1900s and emerged in the early 2000s as a concept to be interconnected in multiple health domains. The questions dominating its broad use and applicability tend to focus on how to define resilience. In 2019, the Trans-NIH Resilience Working Group was formed with a goal to develop an NIH-wide definition of resilience and to achieve consistency and harmony on the design and reporting of resilience research studies. In 1993, an introductory manuscript to a special issue published on the science of resilience included a quote stating, “resilience is at risk for being viewed as a popularized trend that has not been verified through research and is in danger of losing credibility within the scientific community”. The authors of the manuscript also warned against definitional diversity with respect to measures of resilience and urged researchers to clearly operationalize the defi- nition of resilience in all research reports. Remarkably, this call to action served as a primary aim of the Trans-NIH Resilience Working Group when it was organized >25 y after the 1993 special issue on resilience. One of the first activities of the Trans-NIH Resilience Working Group was to host a workshop, in March 2020, which led to the development of a definition of resilience and a conceptual infographic. The definition was intended to be applicable and useful across multiple domains, and it states that resilience encompasses “A system’s capacity to resist, recover, grow, or adapt in response to a challenge or stressor” . A system can represent different domains, levels, and/or processes. Over time, a system’s response to a challenge might show varied degrees of reactions that likely fluctuate in response to the severity of the challenge, the length of time exposed to the challenge, and/or innate/intrinsic factors. To show applicability of the definition in resilience research studies, the Resilience Research Design Tool was later developed to help improve consistency in resilience research reports and to facilitate harmony with respect to measures of resilience outcomes. One of the goals of the resilience framework is to reframe the way we ask research questions, particularly about nutritional interventions like dietary supplements, so that we can better understand health outcomes that are not based solely on disease end points. Going forward, as researchers across various scientific domains and sectors come closer to a unified definition of resilience and perhaps agree to the use of a standard checklist for designing and reporting on resilience studies, there is greater opportunity to harmonize the science and develop more empirical evidence of resilience outcomes.Optimizing performance also includes building resilience in order to enhance the ability to perform tasks and ensuring resilience in order to prevent illness, injury, and disease. Within the US Department of Defense, researchers are able to study different models of physical and psychological stress and the application of different nutritional interventions with Service Members throughout their careers. Various models of stress are introduced, including initial military training , advanced military training courses , service academies , and extreme environments , along with examples of various interventions and outcome measures collected to date. The importance of nutrition on readiness and resilience was identified in military populations more than a decade ago and continues to be of interest. Two specific examples are provided to further explore nutrition interventions aimed at optimizing performance in the Department of Defense. The first, a completed double-blind, randomized, placebo-controlled trial, used a calcium and vitamin D fortified food product to optimize bone health during initial military training of Marine Corps recruits. Using a supplement or food intervention for calcium and vitamin D, participants received 2000-mg calcium and 1000-IU vitamin D per day. The primary outcomes of the study showed that bone markers and vitamin D status improve, but the supplementation did not affect skeletal parameters.