FgPMA1-dsRNA防治小麦赤霉病的研究

doi:10.1016/j.jia.2023.11.046

摘要/Abstract

摘要：

小麦赤霉病（Fusarium head blight）是一种世界性的病害，在全球小麦产区广泛发生，造成严重的经济损失，并且小麦赤霉病菌会产生真菌毒素污染小麦，降低小麦品质。由于缺乏抗病品种，防治小麦赤霉病的主要手段是使用杀菌剂。但是，由于化学药剂的长期和大量使用，抗性问题日益严重，因此，筛选具有新型作用机制的药剂十分重要。前期研究发现禾谷镰刀菌中的质膜ATP酶FgPMA1与氰烯菌酯的靶标肌球蛋白互作，并且各项生命活动中发挥许多重要的功能，因此具有成为靶标的潜力。近年来，研究发现植物与病原菌之间存在一种新的沟通机制，称为跨物种RNA干扰（RNA interference, RNAi），目前，许多研究发现根据RNAi机制开发的核酸农药可以用于有害生物的防治。因此，本研究利用RNAi的方法探究了FgPMA1作为靶标防治小麦赤霉病的可能性。通过将FgPMA1基因划分为6段（PMA1RNAi-1，-2，-3，-4，-5，和-6），并构建了干扰载体及干扰突变体，发现FgPMA1RNAi-1，-2，和-5突变体中FgPMA1的表达量显著降低，菌丝生长受到抑制；产子囊壳能力出现缺陷；分生孢子的产量及形态等受到抑制；毒素的合成受到抑制；致病力下降。通过表型的筛选，我们发现PMA1RNAi-1，-2和-5这三个区段能够起到干扰效果。因此，我们在体外的条件下合成了以这三个区段为模板的dsRNA。在离体条件下，当用25 ng µL^-1 PMA1RNAi-1，-2和-5的dsRNA处理后，禾谷镰刀菌PH-1、亚洲镰刀菌2021和氰烯菌酯抗性菌株YP-1的菌丝生长受到抑制，菌丝中出现膨大的畸形结构，并且失去了产孢能力。在田间，当用PMA1RNAi-1，-2和-5的dsRNA处理麦穗后，禾谷镰刀菌侵染的病斑长度显著下降。这些结果表明：FgPMA1在禾谷镰刀菌中可以作为一个新型的药剂靶标，并且在田间直接施用FgPMA1的dsRNA可以减轻病害的发生。

Abstract:

Fusarium graminearum is a fungal plant pathogen which causes Fusarium head blight (FHB), a devastating disease on cereal crops. Here we report that FgPMA1 could be a new target to control FHB by the application of double-stranded RNA (dsRNA) of FgPMA1. FgPMA1 was divided into 6 segments to generated RNA interference (RNAi) constructs (FgPMA1RNAi-1, -2, -3, -4, -5, and -6), and these constructs were transformed in F. graminearum strain PH-1. The expression of FgPMA1 reduced by 18.48%, 33.48% and 56.93% in FgPMA1RNAi-1, FgPMA1RNAi-2 and FgPMA1RNAi-5, respectively. FgPMA1RNAi-1, -2, and -5 mutants inhibited fungal development, including mycelium growth, mycelial morphology, asexual and sexual development, and toxin production. The length of lesions on wheat leaves, wheat coleoptiles and wheat ears were shorter after infection with FgPMA1RNAi-1, -2, and -5 mutants than wild-type PH-1. These results showed that three segments (FgPMA1RNAi-1, -2, and -5) exhibited effective silencing effects. After treatment with 25 ng µL^-1 dsRNA of these segments in vitro, the growth rate of mycelium growth was significant decreased, mycelium became deformed with bulbous structure at the tip, and the mycelium lost the ability to produce conidia in F. graminearum strain PH-1, Fusarium asiacitum strain 2021 and phenamacril-resistant strain YP-1. After application of FgPMA1RNAi-1-dsRNA and FgPMA1RNAi-2-dsRNA to wheat ears, pathogenicity reduced 34.21-35.40%.

Key words: RNA interference , , FgPMA1 , , Fusarium graminearum , , Nucleic acid pesticide

. FgPMA1-dsRNA防治小麦赤霉病的研究[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2023.11.046.

WU Luo-yu, CHEN Fu-rong, WANG Peng-wei, XU Chong-jing, WEN Wei-dong, HAHN Matthias, ZHOU Ming-guo, HOU Yi-ping. Application of dsRNA of FgPMA1 for disease control on Fusarium graminearum[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2023.11.046.

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