Enhancing tomato disease resistance through endogenous antifungal proteins and introduced nematode-targeting dsRNA of biocontrol agent Bacillus velezensis HS-3

IF 3.8 1区农林科学 Q1 AGRONOMY Pest Management Science Pub Date : 2024-03-21 DOI:10.1002/ps.8087

Juan Han, Jinchi Zhu, Shuyuan Liu, Xuehan Sun, Shunchang Wang, Guopeng Miao

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Abstract

BACKGROUND

As a type of biological control agent (BCA), Bacillus velezensis possesses the efficacy of inhibiting pathogenic microorganisms, promoting plant growth, and overcoming continuous cropping obstacles (CCOs). However, there is limited reporting on the optimization of the cultivation conditions for such biocontrol agents and their role as double-stranded RNA (dsRNA) delivery vectors.

RESULTS

In this study, a Bacillus velezensis strain HS-3 was isolated from the root zone of tomato plants with in vitro anti-Botrytis cinerea activity. The investigation into active compounds revealed that HS-3 predominantly employs proteins with molecular weights greater than 3 kDa for its antifungal activity. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis identified various proteases and chitosanase, further suggesting that HS-3 most likely employs these enzymes to degrade fungal cell walls for its antifungal effect. To optimize the production of extracellular proteins, fermentation parameters for HS-3 were systematically optimized, leading to an optimized medium (OP-M). HS-3 cultured in OP-M demonstrated enhanced capacity to assist tomato plants in withstanding CCOs. However, the presence of excessive nematodes in diseased soil resulted in the disease severity index (DSI) remaining high. An RNA interference mechanism was further introduced to HS-3, targeting the nematode tyrosine phosphatase (TP) gene. Ultimately, HS-3 expressing dsRNA of TP in OP-M effectively assisted tomatoes in mitigating CCOs, reducing DSI to 2.2% and 17.8% of the control after 45 and 90 days of growth, respectively.

CONCLUSION

The advantages of Bacillus velezensis in crop disease management and the mitigation of CCOs become even more pronounced when utilizing both optimized levels of endogenous enzymes and introduced nematode-targeting dsRNA. © 2024 Society of Chemical Industry.

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通过内源抗真菌蛋白和引入生物控制剂 Bacillus velezensis HS-3 的线虫靶向 dsRNA 增强番茄的抗病性。

背景：作为生物防治剂（BCA）的一种，韦氏芽孢杆菌（Bacillus velezensis）具有抑制病原微生物、促进植物生长和克服连作障碍（CCO）的功效。然而，关于优化此类生物控制剂的培养条件及其作为 dsRNA 运送载体的作用的报道却很有限：结果：本研究从番茄植株根区分离出一株 B. velezensis 菌株 HS-3，该菌株具有体外抗灰霉病活性。对活性化合物的研究表明，HS-3 主要利用分子量大于 3 kDa 的蛋白质来发挥其抗真菌活性。LC-MS/MS 分析确定了多种蛋白酶和壳聚糖酶，进一步表明 HS-3 很可能利用这些酶来降解真菌细胞壁，从而产生抗真菌效果。为了优化胞外蛋白的生产，对 HS-3 的发酵参数进行了系统优化，从而产生了一种优化培养基（OP-M）。在 OP-M 培养基中培养的 HS-3 显示出更强的能力，可帮助番茄植株抵御 CCO。然而，病害土壤中线虫过多导致病害严重指数（DSI）居高不下。针对线虫酪氨酸磷酸酶（TP）基因，HS-3 进一步引入了 RNA 干扰机制。最终，在 OP-M 中表达 TP 的 dsRNA 的 HS-3 能有效帮助番茄减轻 CCO，在生长 45 天和 90 天后，DSI 分别降至对照的 2.2% 和 17.8%：结论：当利用优化的内源酶和引入的线虫靶向 dsRNA 时，枯草芽孢杆菌在作物病害管理和缓解 CCO 方面的优势更加明显。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Pest Management Science 农林科学-昆虫学

CiteScore

7.90

自引率

9.80%

发文量

553

审稿时长

4.8 months

期刊介绍： Pest Management Science is the international journal of research and development in crop protection and pest control. Since its launch in 1970, the journal has become the premier forum for papers on the discovery, application, and impact on the environment of products and strategies designed for pest management. Published for SCI by John Wiley & Sons Ltd.