Department of Plant and Microbial Biology, University of California, Berkeley, California, USA.
US Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Mol Plant Pathol. 2023 Sep;24(9):1168-1183. doi: 10.1111/mpp.13361. Epub 2023 Jun 20.
Spray-induced gene silencing (SIGS) is an emerging tool for crop pest protection. It utilizes exogenously applied double-stranded RNA to specifically reduce pest target gene expression using endogenous RNA interference machinery. In this study, SIGS methods were developed and optimized for powdery mildew fungi, which are widespread obligate biotrophic fungi that infect agricultural crops, using the known azole-fungicide target cytochrome P450 51 (CYP51) in the Golovinomyces orontii-Arabidopsis thaliana pathosystem. Additional screening resulted in the identification of conserved gene targets and processes important to powdery mildew proliferation: apoptosis-antagonizing transcription factor in essential cellular metabolism and stress response; lipid catabolism genes lipase a, lipase 1, and acetyl-CoA oxidase in energy production; and genes involved in manipulation of the plant host via abscisic acid metabolism (9-cis-epoxycarotenoid dioxygenase, xanthoxin dehydrogenase, and a putative abscisic acid G-protein coupled receptor) and secretion of the effector protein, effector candidate 2. Powdery mildew is the dominant disease impacting grapes and extensive powdery mildew resistance to applied fungicides has been reported. We therefore developed SIGS for the Erysiphe necator-Vitis vinifera system and tested six successful targets identified using the G. orontii-A. thaliana system. For all targets tested, a similar reduction in powdery mildew disease was observed between systems. This indicates screening of broadly conserved targets in the G. orontii-A. thaliana pathosystem identifies targets and processes for the successful control of other powdery mildew fungi. The efficacy of SIGS on powdery mildew fungi makes SIGS an exciting prospect for commercial powdery mildew control.
喷雾诱导基因沉默（SIGS）是一种新兴的作物害虫防治工具。它利用外源应用的双链 RNA，利用内源性 RNA 干扰机制，特异性降低害虫靶基因的表达。在这项研究中，针对广泛存在的专性生物营养真菌——白粉菌，开发和优化了 SIGS 方法，白粉菌感染农业作物，利用 Golovinomyces orontii-拟南芥病理系统中已知的唑类杀菌剂靶标细胞色素 P450 51（CYP51）。进一步的筛选导致了对白粉菌增殖重要的保守基因靶标和过程的鉴定：凋亡拮抗转录因子在必需的细胞代谢和应激反应中；脂代谢基因脂肪酶 a、脂肪酶 1 和乙酰辅酶 A 氧化酶在能量产生中；以及通过脱落酸代谢（9-顺式-环氧类胡萝卜素双加氧酶、黄烷酮脱氢酶和一种假定的脱落酸 G 蛋白偶联受体）和效应蛋白效应子候选 2 的分泌操纵植物宿主的基因。白粉病是影响葡萄的主要疾病，并且已经报道了广泛的白粉病对应用杀菌剂的抗性。因此，我们为 Erysiphe necator-Vitis vinifera 系统开发了 SIGS，并测试了在 G. orontii-A. thaliana 系统中使用的六个成功靶标。对于所有测试的靶标，在两个系统中都观察到白粉病相似的减少。这表明在 G. orontii-A. thaliana 病理系统中广泛保守靶标的筛选确定了成功控制其他白粉菌的靶标和过程。SIGS 在白粉菌中的功效使 SIGS 成为商业白粉病控制的一个令人兴奋的前景。
