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成功沉默产毒基因并观察到在 VIGS 和 SIGS 实验中毒力降低。

Successful Silencing of the Mycotoxin Synthesis Gene in and Observation of Reduced Virulence in VIGS and SIGS Experiments.

机构信息

Central Research Institute of Epidemiology, Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Novogireevskaya Street 3A, 111123 Moscow, Russia.

Department of Phytopathology, Institute of Phytomedicine, Faculty of Agricultural Sciences, University of Hohenheim, Otto-Sander-Straße 5, 70599 Stuttgart, Germany.

出版信息

Genes (Basel). 2022 Feb 23;13(3):395. doi: 10.3390/genes13030395.

Abstract

Crops constantly experience various biotic stresses during their life cycle, and spp. remain one of the most serious groups of pathogens affecting plants. The ability to manipulate the expression of certain microorganism genes via RNAi creates the opportunity for new-generation dsRNA-based preparations to control a large number of diseases. In this study, we applied virus-induced gene silencing (VIGS), and spray-induced gene silencing (SIGS) to silence the trichothecene-producing gene in as a means to reduce its aggressiveness on spring wheat. Treatment of the fungus with dsTRI5RNA in vitro reduced deoxynivalenol (DON) and 3-acetyldeoxynivalenol (3-A-DON) accumulations by 53-85% and 61-87%, respectively, and reduced expression by 84-97%. VIGS decreased the proportion of infected wheat spikelets by 73%, but upregulation was observed for . SIGS on wheat leaves and ears using certain dsTRI5RNA amounts negatively impacted growth. However, when performing in vivo analyses of mRNA levels, the upregulation of the gene was determined in the variants where fungal colonization was restricted, suggesting a compensatory reaction of the pathogen to RNAi.

摘要

作物在其生命周期中不断经历各种生物胁迫,而 spp. 仍然是影响植物的最严重的病原体群体之一。通过 RNAi 操纵某些微生物基因的表达为新一代基于 dsRNA 的制剂控制大量疾病创造了机会。在这项研究中,我们应用病毒诱导的基因沉默 (VIGS) 和喷雾诱导的基因沉默 (SIGS) 来沉默 中的产毒基因 ,以降低其对春小麦的侵袭性。体外用 dsTRI5RNA 处理真菌可使脱氧雪腐镰刀菌烯醇 (DON) 和 3-乙酰脱氧雪腐镰刀菌烯醇 (3-A-DON) 的积累分别减少 53-85%和 61-87%,并使 的表达减少 84-97%。VIGS 将受感染的小麦小穗比例降低了 73%,但 的表达上调。在小麦叶片和穗上用一定量的 dsTRI5RNA 进行 SIGS 会对 的生长产生负面影响。然而,当对体内 的 mRNA 水平进行分析时,在真菌定植受到限制的变体中确定了该基因的上调,这表明病原体对 RNAi 有补偿反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fbca/8953179/51aef77aa90f/genes-13-00395-g001.jpg

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