Charles Opperman

These studies will enable us to determine specific formulations of nCAP-PGMs that will promote growth in lowland and upland switchgrass varieties.

Overview Wrap and Plant Technology (WPT) involves the conversion of fiber from banana plants into a solid phase matrix used to formulate pesticides. This matrix has been demonstrated to be more efficacious in delivering nematicides for crop protection than current commercial formulations. It is a sustainable environmentally friendly method that reduces the amount of chemicals needed to control pests and increase crop yields. The intended customers are pesticide manufacturers developing new products that contain reduced amounts of active ingredients used in agriculture while achieving the same or greater efficacy. In addition, field trials have determined that WPT can enhance the performance of agrobiological microbes providing an additional application for this material. This disruptive technology places an active ingredient in the right place at the right time and has the potential be a financial success as well as having a positive impact on the environment. Intellectual Merits WPT is a proprietary solid phase matrix for the formulation of agrochemicals. For every pesticide there are two components: the Active Ingredient (AI) that affects the target pathogen and the formulation. The formulation is the carrier that increases solubility, stability, shelf life, UV protection and determines the method of delivery. We have demonstrated that AI formulated with WPT are able to provide improved pest control at lower concentration of same AI than current application rates used by growers. WPT���s increased efficacy of delivering pesticides provides value to the agrochemical manufacturer customer and end user farmer. A prototype using this technology has been used in over 100 field trials over seven years. In over 90% of the trials, the protype increased yield and marketability as compared to current farmer practice. The results of this technology have been reported in multiple conference presentations and peer-reviewed manuscripts. For one nematode species (potato cyst nematode), it was demonstrated that WPT has a novel mechanism of action. Potato cyst nematode is a major pathogen in Europe, Africa and the USA. An International PCT/US2021/071810 has been filed as a result of this work. To date six peer review articles have been published. Broader Impacts Field trials demonstrate WPT reduces the amount of pesticides needed for efficacious crop protection. The global impact of this feature is that less toxic compounds will be used and introduced into the environment. This will reduce the amount of environmental contamination caused from use of pesticides. Another impact for WPT is derived from its use of banana fiber. Banana plants have a 3-year growth cycle. After producing a banana crop the plant dies. Typically, the plant is not recycled, releasing excess nutrients that pollute the environment. By processing banana plants, the excess nutrients can be gathered as a useful fertilizer and the fiber refined which extends the length of the plants��� carbon cycle, reducing carbon pollution. Another impact of reducing the amounts of pesticides used is increased safety to farm workers. The conditions associated with pesticide exposure include hematological alterations, respiratory issues, endocrine dysfunction, neurotoxicity, infertility and, most concerningly, an increased risk of some types of cancer. The use of pesticides and their impact on the health of agricultural workers is a global concern.

Smallholder farming practices in sub-Saharan Africa (SSA) include land-raised seed (piece) use, continuous cultivation (often monoculture) with limited inputs, and virtually non-existent seed (piece) treatment techniques. Yam (Dioscorea spp.) is the primary example of this cropping system and is an extremely important and valuable crop for smallholder farmers in SSA. These practices result in nutrient-depleted soils, nematode infestation, and ultimately low crop yields. Reducing the nematode population in crop soil will dramatically increase crop yield and quality. In many cases, smallholder farmers in SSA lose greater than 50% of their crop to plant-parasitic nematodes, primarily due to lack of available and affordable control options. For this BMG GCE Phase III proposal, we will build upon our promising results from Phase II trials, including increased yields and higher tuber quality and storability for both yam and potato, and use our expertise and connections for pulping banana fiber to validate and prepare for commercialization a developing-world transferable product platform enabling a field deployable paper-like seed (piece) treatment to combat plant pathogenic nematodes. In addition, this platform will be amenable to delivering other crop production moieties, including natural products and oils, necessary minerals and nutrients, or insecticides and fungicides, with its application. Our laboratories������������������ expertise in nematology and lignocellulosic fibrous materials enables us to target the delivery of beneficial small molecules during seed (piece) germination and plant establishment. Importantly the incorporation of active ingredients into a lignocellulose matrix, such as banana tissue paper, allows for widespread distribution of crop protection agents without interfering in smallholder farming practices. The shelf-stable light-weight banana tissue paper can be applied at the point of seed (piece) planting where farmers can use the concept of ����������������wrap and plant��������������� with their own seeds/pieces. Our ���������������wrap and plant������������������ product will be an ����������������active��������������� paper sheet pretreated with ultralow concentrations of active nematicidal ingredient to simply wrap and protect the seed (piece) at planting. The localization of active ingredients carried directly within the paper targets specific plant pathogenic nematodes versus beneficial organisms. Nematodes are primarily a seedling disease, so protection early is critical to the success of the crop, although post harvest losses do occur in yam due to the yam nematode (Scutellonema bradys). Reduction in nematode populations by deploying the ���������������wrap and plant������������������ product protects yam from significant infections that may lead to these post-harvest losses. Our ultimate goal is to validate the ���������������wrap and plant������������������ product in Phase III and to translate the product manufacture to a regional African company for commercialization and distribution.

Banana and plantains (Musa spp.) are major sources of carbohydrates throughout East Africa, especially in the cool highland areas (Gaidashova et al., 2010). In Tanzania, issues such as poor soils, pests and diseases mean that the average yield of banana is only around 9 t. ha-1. However, with pest control and improved management, a yield of 30 t. ha-1 is possible (Mbwana et al,. 1994). One of the major pests affecting banana production are plant parasitic nematodes (PPN), where one species of which alone can cause yield losses of more than 50%, (Speijer & Kajumba, 2000, Atkinson, 2003). Severe infection of banana causes toppling, delayed flowering and reduced yields (Qu��������n��������herv�������� et al., 2012). Plant parasitic nematodes were first reported in Tanzania in 1969 (Bridge 1988). Since then, however, the number of genera/species of PPN found in the country has increased and PPN species have spread to new areas. This is having a major impact, as migratory endoparasitic nematodes have caused banana production in the East African highlands to decline from 8t/ha in 1970 to less that 6t/ha in 1990s (Gold et al, 1999). Moreover, in the Kagera region of Tanzania, between 25 and 95% of bananas sampled from more than 50 farms were found to be impacted by nematode infections (Bridge, 1988). In Tanzania, there is scarce information on important banana nematodes such as species, spread and yield losses among agro-ecological regions. Although a number of nematode species attack banana the most destructive are Pratylenchus spp. (primarily P. goodeyi and P. coffeae), Radopholus similis, and Helicotylenchus spp. (Gowen et al.,2005, Elsen et al., 2004). Previous studies reveal that in Tanzania migratory endoparasitic nematodes, principally R. similis (Cobb, 1893, Thorne, 1949) and P. goodeyi are widespread (Walker et al 1984). Plant parasitic nematodes in banana were first reported in Tanzania in 1969 when Radopholus similis was isolated from diseased materials (Bridge 1988). However, experience shows that from region to region the number of genera have been varying over time and spreading to new areas. Moreover, with climate change the pathogenicity and aggressiveness of strains may also vary. The status of P. coffeae, a widespread and devastating pest of bananas in Latin America and other closely related species such as P. thornei, is unknown in Tanzania. The scant information on nematodes in Tanzania has been recently highlighted in a review by Coyne (2009) and Blomme et al. (2013). Pratylenchus goodeyi is indigenous to Tanzania whereas R. similis is an introduced pest and appears to be currently confined to the humid lowlands. Banana contributes to food and income for small scale farmers in Kagera, Kilimanjaro, Mbeya and Zanzibar, where more than 30% of the total population rely on banana as a staple food crop for their nutrition. Some management efforts (such as crop rotation and use of nematicides) have previously been implemented to control nematodes with varying success. However the overall success has been low and farmers continue to suffer losses causing a shift from their traditional food due to shortage of banana. This threat to food security makes it urgent that we find a nematode management solution. This work will reduce food insecurity to Tanzanians, not only those who rely on banana as a staple food but also others who are in the large majority that prefer banana as food. The benefit and impact of this project will be substantial to countries like Uganda, Rwanda and in West Africa where banana is one of the major food and commercial crops. Diversity within and between populations of the burrowing and lesion nematodes affecting banana can differ widely. Diagnostics of PPN have continued to rely on morphological methods and the use of microscopies that makes it difficult to determine species and develop management measure for the target nematode species. In Tanzania a detailed study on burrowing and lesion nematodes affecting banana has yet to be conducted. The objectives of this study are to (i) Establish the geographical distribution of key plant parasitic nematodes of banana in major banana growing areas (ii) Characterize the phenotypic and genetic diversity of burrowing and lesion nematodes affecting banana (iii) Identify potential microRNAs

IDEA: Develop and validate a biodegradable cellulose matrix platform technology for seed treatment with active ingredients for crop protection enabled by plant viral nanoparticle and traditional cellulosic pulping processes. Phase 1 of this project demonstrated proof-of-concept for this platform by both demonstrating nematode control efficacy in the tomato test system in growth chamber experiments and establishing a banana pulping strategy to create paper matrices. We also established scientific connections with host communities in sub-Sahran Africa insuring translational implementation of the prototype concept during Phase 2 for abating nematode stresses in subsistence farming that reduce crop yield and quality. GOALS: Our immediate follow-up is to foster the translation of the seed treatment technology by implementing the identified infrastructure for adaption by host communities. This includes workshops with local collaborators demonstrating how the simple act of tearing a section of the active paper, encasing the seed by wrapping, and planting will protect the seed throughout germination. We will also build on our success in pulping banana fibers in Rwanda to implement a low-cost production scheme with locally-relevant materials. This will be done by university-exchange and extension services. As a land-grant university, this know-how of translating high-end technology to developing countries is strongly encouraged and supported, if not expected.

Plant parasitic nematodes are one of the world's major agricultural pests, causing in excess of $125 billion in worldwide crop damage annually. Nematodes are primarily a seedling disease, so protection early is critical to the success of the crop. Traditionally, control has depended on highly toxic contact and fumigant pesticides which have now been restricted or eliminated in the United States by the EPA. Issues such as ground water contamination, mammalian and avian toxicity, and residues in food have caused much tighter restrictions on the use of these agricultural chemicals. Current corporate focus for nematode control in major crop plants (maize, soybean, cotton, etc.) is largely seed treatment approaches that provide early protection, which is critical for nematode management. In theory, seed treatment for nematode control is an excellent idea, but in practice it has been largely ineffectual due to delivery problems post-germination. We propose to solve this problem by concentrating nematicides within plant virus-based nanoparticles (PVN) which would be incorporated into a biodegradable cellulosic matrix as a seed coating. The matrix would dissolve and spread with the growing root bundle providing a zone of protection not available with current seed coatings. Furthermore, this approach enables a reduction in the amount of active ingredient applied, exposes the target organism to a much higher concentration of active ingredient, and substantially reduces non-target effects and environmental impact. This proposal is a significant advancement of a project currently funded by the Bill & Melinda Gates Foundation Grand Challenges Explorations Round 8 (Field-Deployable Nutrient-Rich Matrix for Crop Protection). It differs from the current project by testing a larger range of nematicides and crop species specific for the United States (vs. Africa), focusing on first world cropping systems, while also seeking to determine the fate of the PVN within the environment. The overall goal of the project is the development and characterization of PVNs to enhance the performance of a range of nematicides while reducing their environmental impact through decreases in application levels. Specifically, we will: 1) Determine the ability of PVNs to cargo nematicides. We will test a broad range of therapeutically approved nematicides (including ones not currently used for agricultural purposes) for their ability to be loaded onto and released from PVNs. Bioassays using formulated PVNs will be performed on the model nematodes Caenorhabditis elegans and Meloidogyne hapla. Prior to application in soil tests, deactivation procedures for the viral genome will be assayed for their effect(s) on viral infectivity, PVN integrity and nematicide loading and release. 2) Characterize changes in nematicide properties within loaded PVNs in soil. The concentration of nematicides within the PVN will undoubtedly alter their properties with respect to solubility, mobility, persistence and minimum effective dosage. These differences will be assayed by comparing free nematicide to PVN bound nematicide in various soil types via flowthrough assays. Bioassays will then be performed on the collected fractions. 3) Develop and optimize cellulose matrices of various compositions for PVN uptake/release. By controlling packing density via fiber diameter and PVN concentration, locality of the PVN distribution throughout the fiber, biodegradability of the matrix, and functionalization protocol, we will understand the enabling features of the PVN coupled with its carrier matrix. Ultimately, this project will produce a seed treatment that is both efficacious and economical that would both revolutionize and enhance the nematicide market while achieving a crucial movement toward environmentally sustainable agriculture.

We propose to screen a variety of corn lines for resistance to sting nematodes at both juvenile and adult plant stages. We hope to identify germplasm with useful sting nematode resistance.

In this proposed project, we aim to support ongoing and future bioinformatic annotation and curation efforts of the striped bass (Morone saxatilis) and white bass (M. chrysops) genome sequence assemblies, principally the means to make these important resources available online to the research community and to the public in an easily searchable and user friendly format.

We propose to develop an assay for sting nematode resistance and use it to screen at least 300 maize lines. This will constitute a preliminary study for a larger effort to identify and utilize sting nematode resistance.

We propose to purchase an ABI SOLiDTM sequencing platform with ancillary equipment for sample preparation and array-based targeted genomic capture. The equipment brings deep-read sequencing technology to NCSU, which is needed for continued competitiveness and research productivity. It will be housed with other core genomic equipment in NCSU's Genome Science Lab (GSL). The new equipment will serve a diverse user community and will be an essential tool in research requiring i) full genome and targeted resequencing, ii) SNP detection, genotyping, genetic mapping, iii) small RNA discovery, and iv) transcriptional profiling. Research projects directly benefiting from the acquisition of the ABI SOLiDTM focus on understanding the relationship between genetic variation and phenotypic diversity and range from understanding how life persists in extreme environments to determining the mechanisms by which adaptive variation in butterfly wing patterns arise. The acquisition of the ABI SOLiDTM will be coupled with the development of targeted genomic capture methodologies and the establishment of core bioinformatics support for the GSL community. This combination of state-of-the-science equipment, technical expertise, and informatic support, makes the GSL unique among core genomic facilities, facilitates technology transfer among our diverse user community, and provides novel training experiences for our undergraduate and graduate students and postdoctoral researchers.