中国农业科学 ›› 2019, Vol. 52 ›› Issue (24): 4505-4517.doi: 10.3864/j.issn.0578-1752.2019.24.006

• 植物保护 • 上一篇    下一篇

西兰花残体还田对棉花黄萎病防治效果及其对不同生育时期土壤细菌群落的影响

赵卫松,郭庆港,李社增,王亚娇,鹿秀云,王培培,苏振贺,张晓云,马平()   

  1. 河北省农林科学院植物保护研究所/河北省农业有害生物综合防治工程技术研究中心/农业农村部华北北部作物有害生物综合治理重点实验室, 河北保定 071000
  • 收稿日期:2019-06-03 接受日期:2019-06-28 出版日期:2019-12-16 发布日期:2020-01-15
  • 通讯作者: 马平
  • 作者简介:赵卫松,Tel:0312-5927076;E-mail:zhaoweisong1985@163.com。
  • 基金资助:
    国家重点研发计划(2017YFD0200601);国家公益性行业农业科研专项(201503109);国家棉花产业技术体系(CARS-15-17)

Control Efficacy of Broccoli Residues on Cotton Verticillium Wilt and Its Effect on Soil Bacterial Community at Different Growth Stages

WeiSong ZHAO,QingGang GUO,SheZeng LI,YaJiao WANG,XiuYun LU,PeiPei WANG,ZhenHe SU,XiaoYun ZHANG,Ping MA()   

  1. Plant Protection Institute of Hebei Academy of Agricultural and Forestry Sciences/IPM Centre of Hebei Province/Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture and Rural Affairs, Baoding 071000, Hebei
  • Received:2019-06-03 Accepted:2019-06-28 Online:2019-12-16 Published:2020-01-15
  • Contact: Ping MA

摘要:

【目的】研究土壤中添加西兰花残体(broccoli residues,BR)对棉花黄萎病及不同生育时期土壤细菌群落结构的影响,为棉花黄萎病的绿色生态防控和化学农药的减量施用提供新途径和思路。【方法】通过田间小区试验,分别以西兰花残体还田和不含有西兰花残体(CK)的耕层土壤中播种的棉花为试验对象,监测不同处理的棉花黄萎病的发生动态。采用实时荧光定量PCR(real-time PCR)和高通量测序(Illumina MiSeq)技术分别测定不同时期土壤中大丽轮枝菌(Verticillium dahliae)DNA拷贝数量和土壤细菌群落,进而分析西兰花残体还田对土壤中病原菌数量变化和土壤细菌群落的影响。采用主成分分析探索西兰花残体还田对不同生育时期土壤细菌群落在门和属水平的结构变化特征与规律。【结果】西兰花残体还田后棉花黄萎病的发病率和病情指数均呈现出下降趋势,且发病高峰时间推迟,在发病高峰期对黄萎病的防治效果达到70.77%,建立了整个生育期内病情指数-时间病程发展曲线,平均防治效果为57.21%。与空白对照相比,西兰花残体还田处理在棉花生长季和非生长季的4个阶段土壤中大丽轮枝菌DNA拷贝数量分别降低了10.96%、11.11%、25.95%和11.25%。高通量测序分析表明,不同生育时期西兰花残体还田处理显著提高了土壤细菌多样性,丰富度指数Chao1均显著上升,ACE指数在播种前期、花铃期和吐絮期显著上升。与空白对照相比,在不同生育时期内,西兰花残体还田后土壤中的放线菌门(Actinobacteria)菌群的相对丰度呈现显著上升。同时研究表明,西兰花残体还田后优势菌群组成在不同生育时期存在差别,在播种前期和苗期,厚壁菌门(Firmicutes)菌群上升为优势菌群;在蕾期和花铃期,蓝藻细菌门(Cyanobacteria)菌群成为优势菌群。主成分分析表明,西兰花残体还田改变了苗期、蕾期、花铃期和吐絮期的细菌群落结构。进一步分析表明,西兰花残体还田显著增加了链霉菌属(Streptomyces)和芽孢杆菌属(Bacillus)菌群的相对丰度。【结论】西兰花残体还田能够有效抑制棉花黄萎病的发生,降低土壤中大丽轮枝菌DNA拷贝的数量,改变土壤细菌群落结构,增加有益微生物的相对丰度,是一种有效控制棉花黄萎病的绿色措施。

关键词: 西兰花残体, 大丽轮枝菌, 棉花黄萎病, 细菌群落结构, 高通量测序, 生育时期

Abstract:

【Objective】The objective of this study is to research the effects of broccoli residues (BR) on cotton verticillium wilt and soil bacterial community structure at different growth stages, and to provide new ways and ideas for green ecological control of cotton verticillium wilt and reduction of chemical pesticides.【Method】In the field plot experiment, the soil planted with cotton was treated with broccoli residues and no broccoli residues (CK). The incidence dynamics of cotton verticillium wilt in different treatments were monitored at different growth stages. The number of DNA copies of Verticillium dahliae was determined by real-time quantitative PCR, and the structure of soil bacterial communities was measured by high-throughput sequencing (Illumina MiSeq). The effects of broccoli residues returning to soil on the quantity of pathogen and the community of soil bacteria were analyzed. Principal component analysis (PCA) was used to explore the characteristics and rules of structural changes of soil bacterial community at the level of phylum and genus in different growth stages.【Result】The incidence and disease index of cotton verticillium wilt showed a downward trend after the return of broccoli residues to soil, respectively. Moreover, the peak period of verticillium wilt was delayed, and 70.77% control efficacy was achieved at the peak period. The development curve of disease index and time course of disease in the whole growth period was established, and the average control efficacy was 57.21%. Compared to the blank control, the number of DNA copies of V. dahliae was decreased by 10.96%, 11.11%, 25.95% and 11.25% at the 4 stages of cotton growing and non-growing season, respectively, after the return of broccoli residues to soil. Illumina MiSeq analysis showed that broccoli residues treatment significantly increased soil bacterial diversity. The richness index Chao1 was increased significantly at all growth stages, and the ACE index was significantly increased at the pre-sowing stage, flowering and boll stage and boll-opening stage. Compared to the blank control, the relative abundance of Actinobacteria was increased significantly at different growth stages after the return of broccoli residues to soil. Meanwhile, the results showed that there were differences in the composition of dominant microorganisms at different growth stages after the return of broccoli residues to soil. For example, Firmicutes increased to the dominant microflora at the pre-sowing stage and seedling stage, and Cyanobacteria became the dominant microflora at the bud stage and flowering and boll stage. Principal component analysis showed that broccoli residues to soil changed the structure of bacterial community at seedling stage, bud stage, flowering and boll stage and boll-opening stage. Further analysis showed that the relative abundance of Streptomyces and Bacillus was significantly increased after the return of broccoli residues to soil.【Conclusion】Broccoli residues returning to soil can effectively inhibit the occurrence of cotton verticillium wilt, reduce the number of DNA copies of V. dahliae in soil, change the structure of soil bacterial community, and increase the relative abundance of beneficial microorganisms, which is an effective green measure to control cotton verticillium wilt.

Key words: broccoli residues, Verticillium dahliae, cotton verticillium wilt, bacterial community structure, Illumina MiSeq, growth stage