中国农业科学 ›› 2022, Vol. 55 ›› Issue (9): 1781-1789.doi: 10.3864/j.issn.0578-1752.2022.09.007

• 植物保护 • 上一篇    下一篇

亚油酸乙醇胺诱导番茄对灰葡萄孢抗性的作用及机制

邵淑君(),胡璋健,师恺*()   

  1. 浙江大学农业与生物技术学院,杭州 310058
  • 收稿日期:2021-11-06 修回日期:2021-12-10 出版日期:2022-05-01 发布日期:2022-05-19
  • 通讯作者: 师恺
  • 作者简介:邵淑君,E-mail: ssjun@zju.edu.cn
  • 基金资助:
    国家自然科学基金(32172650);浙江省重点研发计划(2021C02040)

The Role and Mechanism of Linoleyl Ethanolamide in Plant Resistance Against Botrytis cinerea in Tomato

SHAO ShuJun(),HU ZhangJian,SHI Kai*()   

  1. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058
  • Received:2021-11-06 Revised:2021-12-10 Online:2022-05-01 Published:2022-05-19
  • Contact: Kai SHI

摘要:

【背景】灰葡萄孢(Botrytis cinerea)引起的灰霉病是危害番茄(Solanum lycopersicum)的重要病害之一,防治不及时可造成30%—40%减产。目前生产上多以化学防治为主,但存在农产品安全及环境污染的风险。N-酰基乙醇胺(NAE)是植物体内天然存在的一类脂质生物活性化合物,其在哺乳动物中具有多种免疫功能,但其在植物免疫中的作用和机制尚不清楚。【目的】探讨N-酰基乙醇胺在诱导番茄对灰葡萄孢防御中的作用,为研发番茄灰霉病绿色防控技术提供依据。【方法】将灰葡萄孢分别接种在含有硬脂酰乙醇胺(NAE 18:0)、亚油酸乙醇胺(NAE 18:2)、廿二碳五烯酸乙醇胺(NAE 22:5)的培养基上,观察灰葡萄孢的生长情况。在此基础上,以番茄‘Moneymaker’植株为材料,硬脂酰乙醇胺、亚油酸乙醇胺、廿二碳五烯酸乙醇胺外源处理番茄叶片后接种灰葡萄孢,统计病情指数,测定荧光参数。采用qRT-PCR技术明确亚油酸乙酰胺处理后番茄叶片灰葡萄孢Actin的相对表达量,进一步测定番茄叶片中主要抗病基因PI IPR-1NPR1NrACO1、PYR1a等的相对表达量及茉莉酸(jasmonic acid,JA)、水杨酸(salicylic acid,SA)、乙烯(ethylene,ETH)、脱落酸(abscisic acid,ABA)、生长素(indoleacetic acid,IAA)含量。并以乙烯信号转导突变体植株never ripenr)及对照植株Pearson(PB)为材料,在外源亚油酸乙酰胺处理后接种灰葡萄孢,测定番茄叶片叶绿素荧光参数和灰葡萄孢Actin的相对表达量。【结果】体外培养试验结果表明灰葡萄孢生长并不受外源N-酰基乙醇胺的影响,外源施用N-酰基乙醇胺能显著提高番茄植株对灰霉病的抗性,缓解由灰葡萄孢侵染导致的番茄叶片光系统II实际光化学效率(ΦPSII)的下降。3种N-酰基乙醇胺中,亚油酸乙醇胺施用后番茄叶片灰霉病病情指数下降最为明显,灰葡萄孢Actin的相对表达量下调60%,其诱导灰霉病抗性效果最佳。番茄植株接种灰葡萄孢后抗性基因PI IPR-1NPR1、NrACO1的表达水平均有不同程度上调。外源亚油酸乙醇胺处理使得番茄响应灰葡萄孢接种后PI INrACO1的表达进一步增强,其中乙烯合成基因ACO1的表达水平最高。番茄植株接种灰葡萄孢后叶片水杨酸、茉莉酸、生长素和乙烯含量增加,但外源亚油酸乙醇胺处理并接种灰葡萄孢后只有乙烯含量显著增加。进一步研究发现,番茄乙烯突变体nr植株中,外源亚油酸乙醇胺对灰葡萄孢的抗性诱导作用显著受到抑制。【结论】外源施用亚油酸乙醇胺能够提高番茄内源光合效率和抗病基因的表达及内源激素乙烯的含量,增强番茄植株对灰霉病的抗性,推测其诱导抗性作用可能与乙烯信号路径相关。

关键词: 番茄, N-酰基乙醇胺, 亚油酸乙醇胺, 灰霉病, 灰葡萄孢, 乙烯

Abstract:

【Background】Gray mold caused by Botrytis cinerea is one of the important diseases of tomato and causes significant yield losses up to 30%-40%. Nowadays, chemical pesticide is usually used in tomato production, which is effective but increases the risk of food safety and results in environmental pollution. N-acylethanolamines (NAEs) are a kind of naturally lipid bioactive compounds in plants, which have been identified to have a variety of immune functions in mammals, however, its function and the underlying mechanism in plant immunity are still unclear.【Objective】The objective of this study is to investigate the effects of NAEs on tomato plant defense against B. cinerea infection, and to provide a basis for the development of green control technology of tomato gray mold.【Method】The B. cinerea was cultured in medium containing NAE 18:0, NAE 18:2, NAE 22:5, respectively, to evaluate their effects on B. cinerea growth. Tomato ‘Moneymaker’ plants were infected by B. cinerea with or without exogenous NAE 18:0, NAE 18:2, NAE 22:5, and disease index and fluorescence parameters of tomato leaves were measured. qRT-PCR was used to analyze the relative gene expression of B. cinerea Actin in tomato leaves that infected by B. cinerea with or without NAE 18:2 treatment. Transcript abundance of defense-related genes (e.g. PI I, PR-1, NPR1, Nr, ACO1, PYR1a), and contents of plant hormones (e.g. JA, SA, ETH, ABA, IAA) were measured. Fluorescence parameters of tomato leaves and the relative gene expression of B. cinerea Actin were analyzed in ethylene-insensitive mutant infected by B. cinerea with NAE 18:2.【Result】The growth of B. cinerea was not affected by exogenous NAEs treatment during in vitro culture. Exogenous application of NAEs could significantly improve the resistance of tomato plants to B. cinerea, and alleviate the decrease of photosystem II photochemical efficiency (ΦPSII) caused by B. cinerea infection. NAE 18:2 had the best effect on tomato plant defense against B. cinerea among the NAEs, which obviously reduced the disease index and the Actin transcript level of B. cinerea by 60%. The expression levels of PI I, PR-1, NPR1, Nr and ACO1 could be induced by B. cinerea infection but not by NAE 18:2 treament. The expression levels of PI I, Nr and ACO1 were up-regulated when plants were pre-treated by NAE 18:2 before B. cinerea infection, and the expression level of ACO1 was the highest. Compared to the control, the contents of SA, JA, IAA and ETH in the leaves were increased significantly after B. cinerea infection, while only the contents of ETH were further increased when pre-treated by NAE 18:2. Moreover, exogenous NAE 18:2 pre-treatment could not improve the defense against B. cinerea in the ETH-insensitive mutant nr.【Conclusion】Exogenous NAE18:2 treatment can increase leaf photosynthesis, transcript abundance of defense-related genes, and the content of plant hormone ETH. It induce the resistance of tomato plants to gray mold, which may depend on the ETH signaling pathway.

Key words: tomato, N-acylethanolamines (NAE), linoleyl ethanolamide (NAE 18:2), gray mold, Botrytis cinerea, ethylene (ETH)