温度对棉花黄萎病发生及寄主防御反应的影响

doi:10.3864/j.issn.0578-1752.2023.23.009

中国农业科学 ›› 2023, Vol. 56 ›› Issue (23): 4671-4683.doi: 10.3864/j.issn.0578-1752.2023.23.009

温度对棉花黄萎病发生及寄主防御反应的影响

张亚林¹^,²(), 蒋艳², 赵丽红², 冯自力², 冯鸿杰², 魏锋², 周京龙², 朱荷琴²(), 马峙英¹()

¹ 河北农业大学农学院/华北作物改良与调控国家重点实验室/河北省作物种质资源重点实验室，河北保定 071000
² 中国农业科学院棉花研究所/棉花生物育种与综合利用全国重点实验室，河南安阳 455000

收稿日期:2023-08-02 接受日期:2023-09-03 出版日期:2023-12-04 发布日期:2023-12-04
通信作者:
朱荷琴，E-mail：heqinanyang@163.com。
马峙英，E-mail：mzhy@hebau.edu.cn
联系方式: 张亚林，E-mail：zhangyalin@caas.cn。
基金资助:
国家自然科学基金(32201752); 第七师胡杨河市财政科技计划项目(2023C07); 中国农业科学院科技创新工程(CAAS-ASTIP-ICR)

Effect of Temperature on the Occurrence of Cotton Verticillium Wilt and Host Defense Response

ZHANG YaLin¹^,²(), JIANG Yan², ZHAO LiHong², FENG ZiLi², FENG HongJie², WEI Feng², ZHOU JingLong², ZHU HeQin²(), MA ZhiYing¹()

¹ College of Agronomy, Hebei Agricultural University/State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory for Crop Germplasm Resources of Hebei, Baoding 071000, Hebei
² Institute of Cotton Research, Chinese Academy of Agricultural Sciences/National Key Laboratory of Cotton Bio-Breeding and Integrated Utilization, Anyang 455000, Henan

Received:2023-08-02 Accepted:2023-09-03 Published:2023-12-04 Online:2023-12-04

摘要/Abstract

摘要：

【目的】解析不同温度对棉花黄萎病发生的影响以及调控寄主防御反应的机理，揭示温度对病原菌和寄主的双重作用，为该病害的绿色防控和温度调控研究提供理论依据。【方法】以棉花抗病品种中植棉2号（ZZM2）、感病品种冀棉11（JM11）为试验材料，采用室内实验与病圃实验相结合，设置恒定温度（22、25、28和32 ℃）、自然变温两个处理，检测温度对大丽轮枝菌（Verticillium dahliae）生长、侵染定殖、棉花黄萎病发生的影响；通过调查活性氧（ROS）爆发、过氧化氢（H₂O₂）含量、胼胝质积累、防御相关基因表达水平等植物防御相关指标，解析温度调控棉花寄主防御反应的机制。【结果】在培养基上，25 ℃是大丽轮枝菌菌丝生长最适温度，22—28 ℃范围内适宜产孢；与培养基相比，不同棉花品种的叶片提取液对大丽轮枝菌生长均有促进作用，感病品种JM11促进作用更强；当温度为25—28 ℃时，ZZM2和JM11植株黄萎病发生均较重，低温22 ℃、高温32 ℃均不利于黄萎病发生。同时，在25 ℃处理下，棉花中大丽轮枝菌侵染定殖能力强，感病品种JM11比抗病品种ZZM2更易受到大丽轮枝菌的侵染，与棉花黄萎病病情指数的结果基本一致；进一步，温度显著影响了棉花寄主防御反应：与22—28 ℃相比，无论是否接种大丽轮枝菌，ZZM2和JM11在32 ℃处理下产生的ROS爆发更强烈；在25 ℃处理下，ZZM2和JM11叶片中H₂O₂含量最低；在32 ℃处理下，ZZM2和JM11叶片胼胝质积累量较高，分别为817、575个/mm²，是未接菌对照的2.04、1.80倍；棉花叶片中PAL、POD、PPO防御相关基因在25—28 ℃处理下表达量降低，低于22、32 ℃处理。【结论】温度对大丽轮枝菌生长、寄主防御反应具有双重作用，进而影响棉花黄萎病发生。其中无论恒定温度、自然变温，25—28 ℃均有利于大丽轮枝菌在棉花中侵染定殖，显著降低寄主防御反应，导致棉花黄萎病严重发生。

关键词: 温度, 棉花黄萎病, 大丽轮枝菌, 防御反应, 侵染

Abstract:

【Objective】The objective of this study is to analyze the effect of different temperatures on the occurrence of cotton Verticillium wilt and the mechanism of regulating host defense response, reveal the dual effects of temperature on pathogen and host, and to provide a theoretical basis for the green control and temperature regulation of this disease.【Method】Cotton Verticillium wilt resistant variety Zhongzhimian 2 (ZZM2) and susceptible variety Jimian 11 (JM11) were employed as experimental materials, indoor experiments and disease nursery experiments were jointly used to set constant temperatures (22, 25, 28, and 32 ℃) and natural temperature variation to determine the effect of temperature on the growth, infection and colonization of Verticillium dahliae, and the occurrence of cotton Verticillium wilt. The mechanism of temperature regulation of cotton host defense response was analyzed by utilizing indicators such as reactive oxygen species (ROS) outbreak, H₂O₂ content, callose accumulation, and the expression of defense-related genes.【Result】On the culture media, 25 ℃ was the optimal temperature for the mycelial growth of V. dahliae, and the range of 22-28 ℃ was suitable for spore yield. Compared with the culture media, the leaf extracts of resistant variety ZZM2 and susceptible variety JM11 had a promoting effect on the growth of V. dahliae, and JM11 promoted stronger. When the temperatures were between 25 and 28 ℃, both ZZM2 and JM11 suffered from severe Verticillium wilt, and low temperature 22 ℃ and high temperature 32 ℃ were not conducive to the occurrence of Verticillium wilt. Meanwhile, under 25 ℃ treatment, the colonization ability of V. dahliae in cotton was strong, JM11 was more susceptible to V. dahliae infection than ZZM2, which was basically consistent with the results of the cotton Verticillium wilt disease index. Furthermore, temperature significantly affected the host defense response of cotton: compared with 22-28 ℃, whether inoculated with V. dahliae or not, the ROS outbreak produced by ZZM2 and JM11 under 32 ℃ treatment was stronger; under 25 ℃ treatment, the H₂O₂ content in ZZM2 and JM11 leaves was the lowest; under 32 ℃ treatment, the accumulation of callose in ZZM2 and JM11 leaves was higher, which was 2.04 and 1.80 times higher than that of the untreated control, respectively; the expression level of PAL, POD, and PPO defense-related genes in cotton leaves decreased under treatments at 25-28 ℃, which was lower than that under treatments at 22 and 32 ℃.【Conclusion】Temperature has a dual effect on the growth of V. dahliae and host defense response, which in turn affects the occurrence of cotton Verticillium wilt. Regardless of constant temperature or natural temperature variation, 25-28 ℃ are beneficial for the colonization of V. dahliae in cotton, significantly reduce the host defense response, and lead to severe cotton Verticillium wilt.

Key words: temperature, cotton Verticillium wilt, Verticillium dahliae, defense response, infection

张亚林, 蒋艳, 赵丽红, 冯自力, 冯鸿杰, 魏锋, 周京龙, 朱荷琴, 马峙英. 温度对棉花黄萎病发生及寄主防御反应的影响[J]. 中国农业科学, 2023, 56(23): 4671-4683.

ZHANG YaLin, JIANG Yan, ZHAO LiHong, FENG ZiLi, FENG HongJie, WEI Feng, ZHOU JingLong, ZHU HeQin, MA ZhiYing. Effect of Temperature on the Occurrence of Cotton Verticillium Wilt and Host Defense Response[J]. Scientia Agricultura Sinica, 2023, 56(23): 4671-4683.

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基因名 Gene name	引物序列 Primer sequence (5′→3′)
β-tubulin	F: CCGCCGGTCCATCAGTCTCTCTGTTTATAC
β-tubulin	R: CGCCTGCGGGACTCCGAGCGAGCGTAAC
Ubiquitin	F: GAGTCTTCGGACACCATTG
Ubiquitin	R: CTTGACCTTCTTCTTCTTGTGC
PAL	F: TGGTGGCTGAGTTTAGGAAA
PAL	R: TGAGTGAGGCAATGTGTGA
POD	F: CCGCATAACCATCACAAG
POD	R: ACTCTCATCACCTTCAACA
PPO	F: ATATCCTTGTTCTGTCTGCTA
PPO	R: CTCCTTCTACCGTCTCTTC

温度对棉花黄萎病发生及寄主防御反应的影响

Effect of Temperature on the Occurrence of Cotton Verticillium Wilt and Host Defense Response

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