Virginia Agricultural Experiment Station

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https://hdl.handle.net/10919/24235
The Virginia Agricultural Experiment Station conducts research on food and fiber systems, their impact on the environment, and their relation to the future needs of Virginia, the nation, and the world.

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Browsing Virginia Agricultural Experiment Station by Subject "0607 Plant Biology"

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    Application of Exogenous dsRNAs-induced RNAi in Agriculture: Challenges and Triumphs
    Das, Protiva Rani; Sherif, Sherif M. (Frontiers, 2020-06-25)
    In recent years, RNA interference (RNAi) machinery has widely been explored by plant biologists for its potential applications in disease management, plant development, and germplasm improvement. RNAi-based technologies have mainly been applied in the form of transgenic plant generation and host-induced-gene-silencing (HIGS). However, the approval of RNAi-based transgenic plants has always been challenging due to the proclaimed concerns surrounding their impacts on human health and the environment. Lately, exogenous applications of double-stranded RNAs (dsRNAs), short interfering RNAs (siRNAs), and hairpin RNAs (hpRNAs) has emerged as another technology that could be regarded as more eco-friendly, sustainable, and publicly acceptable than genetic transformation. Inside the plant cell, dsRNAs can undergo several steps of processing, which not only triggers RNAi machinery but may also involve transitive and systemic silencing, as well as epigenetic modifications. Therefore, along with the considerations of proper exogenous applications of dsRNAs, defining their final destination into plant cells is highly relevant. In this review, we highlighted the significance of several factors that affect dsRNA-induced gene silencing, the fate of exogenous dsRNAs in the plant cell, and the challenges surrounding production technologies, cost-effectiveness, and dsRNAs stability under open-field conditions. This review also provided insights into the potential applications of exogenous dsRNAs in plant protection and crop improvement.
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    Conidial production and viability of Calonectria pseudonaviculata on infected boxwood leaves as affected by temperature, wetness, and dryness periods
    Avenot, Herve F.; Baudoin, Antonius B.; Hong, Chuanxue (Wiley, 2021-10-31)
    Calonectria pseudonaviculata causes lesions on boxwood leaves and twigs. Controlled-environment experiments were conducted to determine the effects of temperature and leaf wetness period on C. pseudonaviculata sporulation on diseased (cv. Suffruticosa) leaves and of dryness periods and high temperature on conidial survival. Infected leaves were incubated in moist chambers and subjected to six temperatures (9, 13, 17, 21, 25, and 29°C) and six leaf wetness periods (0, 12, 24, 40, 48, and 72 h). Spore production was influenced significantly by wetness period, temperature, and their interaction. Increasing duration of leaf wetness and increasing temperature generally increased sporulation, with no sporulation occurring at 29°C or 9 and 13°C, except at 72 h of wetness exposure, while it was optimal at 21°C. Detached leaves with profuse conidia were subjected to a range of drying (relative humidity at 65%) times (0, 2, 4, 6, and 8 h) at two temperatures of 21 and 29°C. Conidia were then harvested and plated on water agar. Germinating conidia were counted to measure the spore viability. Spore mortality increased with increasing dryness duration at both temperatures but occurred more quickly and severely at 29 than 21°C. Overall, this study extended biological knowledge of conditions required for crucial stages of the C. pseudonaviculata disease cycle and the obtained results will be vital for developing boxwood blight forecasting and management tools.
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    Proof of Concept for Shoot Blight and Fire Blight Canker Management with Postinfection Spray Applications of Prohexadione-Calcium and Acibenzolar-S-Methyl in Apple
    Aćimović, Srđan G.; Meredith, Christopher L.; Santander, Ricardo Delgado; Khodadadi, Fatemeh (Scientific Societies, 2021-12-03)
    To reduce the severity of shoot blight and prevent the resulting development of cankers on perennial apple wood, we evaluated eight fire blight postinfection spray programs of prohexadione-calcium (PCA) alone or with acibenzolar-S-methyl (ASM) over 2 years. On mature trees of cultivar Royal Court, a single application of the high PCA rate (247 mg/liter) at 2 to 3 days after inoculation resulted in 89.5 and 69.5% reduction of shoot blight severity after inoculation. Two applications of PCA 247 mg/liter 12 or 14 days apart, with the first one applied 2 to 3 days after inoculation, resulted in 78.8 and 74.5% reduction of shoot blight severity in both years. A 100% control of canker incidence on perennial wood from infected shoots in both years was achieved with a single application of PCA (247 mg/liter) applied at 2 or 3 days after the inoculation, and three applications of PCA (125 mg/liter) + ASM (25 mg/liter) 12 to 16 days apart reduced canker incidence by 83.5 and 69% in the 2 years. The other programs with lower PCA rates and frequencies of application reduced shoot blight severity 50.8 and 51.8% (PCA) and 62.6 to 72% and 59.3% (PCA + ASM) over 2 years, respectively. Reduction of canker incidence on wood by the other programs was 66.5% and 69 to 90.4% in the two years, respectively. As fire blight cankers lead to death of dwarf apple trees and serve as primary sources of inoculum, our effective PCA and PCA + ASM programs could serve as viable postinfection management options. These treatments can reduce or prevent canker development and thus significantly abate tree losses in high-density apple orchards after fire blight epidemics occur.
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    Strain-level identification of bacterial tomato pathogens directly from metagenomic sequences
    Mechan Llontop, Marco Enrique; Sharma, Parul; Aguilera Flores, Marcela; Yang, Shu; Pollock, Jill; Tian, Long; Huang, Chengjie; Rideout, Steven L.; Heath, Lenwood S.; Li, Song; Vinatzer, Boris A. (Scientific Societies, 2019-12-12)
    Routine strain-level identification of plant pathogens directly from symptomatic tissue could significantly improve plant disease control and prevention. Here we tested the Oxford Nanopore Technologies (ONT) MinIONTM sequencer for metagenomic sequencing of tomato plants either artificially inoculated with a known strain of the bacterial speck pathogen Pseudomonas syringae pv. tomato (Pto), or collected in the field and showing bacterial spot symptoms caused by either one of four Xanthomonas species. After species-level identification using ONT's WIMP software and the third party tools Sourmash and MetaMaps, we used Sourmash and MetaMaps with a custom database of representative genomes of bacterial tomato pathogens to attempt strain-level identification. In parallel, each metagenome was assembled and the longest contigs were used as query with the genome-based microbial identification Web service LINbase. Both the read-based and assembly-based approaches correctly identified Pto strain T1 in the artificially inoculated samples. The pathogen strain in most field samples was identified as a member of Xanthomonas perforans group 2. This result was confirmed by whole genome sequencing of colonies isolated from one of the samples. Although in our case, metagenome-based pathogen identification at the strain-level was achieved, caution still needs to be exerted when interpreting strain-level results because of the challenges inherent to assigning reads to specific strains and the error rate of nanopore sequencing.