中國農業科學 ?? 2020, Vol. 53 ?? Issue (5): 942-954.doi: 10.3864/j.issn.0578-1752.2020.05.007

? 植物保護 ? 上一篇    下一篇

花鈴期棉花黃萎病抗病與感病品種對 土壤細菌群落結構的影響

趙衛松,郭慶港,李社增,王培培,鹿秀云,蘇振賀,張曉云,馬平()   

  1. 河北省農林科學院植物保護研究所/河北省農業有害生物綜合防治工程技術研究中心/農業農村部華北北部作物有害生物綜合治理重點實驗室, 河北保定 071000
  • 收稿日期:2019-08-29 接受日期:2019-09-26 出版日期:2020-03-01 發布日期:2020-03-14
  • 通訊作者: 馬平 E-mail:[email protected]
  • 作者簡介:趙衛松,Tel:0312-5927076;E-mail:[email protected]
  • 基金資助:
    國家重點研發計劃(2017YFD0200601);國家公益性行業農業科研專項(201503109);國家棉花產業技術體系(CARS-15-17)

Effect of Wilt-Resistant and Wilt-Susceptible Cotton on Soil Bacterial Community Structure at Flowering and Boll Stage

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

  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:2019-08-29 Accepted:2019-09-26 Online:2020-03-01 Published:2020-03-14
  • Contact: Ping MA E-mail:[email protected]

摘要:

【目的】研究花鈴期棉花黃萎病抗/感品種土壤細菌群落結構,了解抗/感品種土壤細菌群落結構與土壤理化性質之間的關系,為棉花黃萎病的監測與綠色生態防控打下理論基礎。【方法】通過田間小區試驗,以感病品種(鄂荊1號,EJ)和抗病品種(冀863,J863)為試驗材料,采用實時熒光定量PCR(real-time PCR)和高通量測序(Illumina MiSeq)技術分別測定花鈴期不同階段(盛花期、開花后期和結鈴期)土壤中大麗輪枝菌(Verticillium dahliae)數量和土壤細菌群落結構,結合冗余分析(RDA)明確細菌群落結構與土壤理化性質的相關性。【結果】棉花黃萎病的發生與土壤中大麗輪枝菌ITS基因拷貝數量存在不同程度的相關性,其中感病品種EJ的發病率和病情指數與土壤中病原菌數量呈正相關,而抗病品種J863的發病率和病情指數與病原菌數量相關性不大。除盛花期外,棉花開花后期和結鈴期抗病品種J863土壤中的病原菌數量低于感病品種EJ。高通量測序分析表明,除開花后期,抗病品種J863在盛花期和結鈴期的細菌豐富度Chao1和ACE指數均高于感病品種EJ。主成分分析表明,抗/感品種之間及其在花鈴期不同階段的土壤細菌群落結構存在差異。群落組成方面,在門水平上,感病品種EJ的部分優勢菌群平均相對豐度低于抗病品種J863,如放線菌門(Actinobacteria)、芽單胞菌門(Gemmatimonadetes)、綠彎菌門(Chloroflexi)、擬桿菌門(Bacteroidetes)、硝化螺菌門(Nitrospirae)、Patescibacteria和裝甲菌門(Armatimonadetes),降低幅度分別為16.38%、4.05%、2.25%、6.58%、7.10%、20.60%和35.78%;在屬水平上,感病品種EJ的部分優勢菌群平均相對豐度低于抗病品種J863,包括鞘氨醇單胞菌屬(Sphingomonas)、芽單胞菌屬(Gemmatimonas)、BryobacterIamiaPseudarthrobacter、芽球菌屬(Blastococcus)、紅色桿菌屬(Rubrobacter)、類諾卡氏屬(Nocardioides)、Pontibacter、鏈霉菌屬(Streptomyces)、Gemmatirosa、微單孢菌屬(Micromonospora)和Solirubrobacter,下降幅度分別為5.09%、19.41%、13.79%、2.36%、10.78%、34.47%、46.76%、61.84%、52.75%、48.61%、74.79%、9.13%和26.42%。冗余分析(RDA)表明,土壤細菌群落結構受硝態氮(NO3 --N)、速效磷(AP)、銨態氮(NH4 +-N)、無機磷(IP)、pH和有機質(OM)指標影響。【結論】土壤中大麗輪枝菌的數量與棉花抗/感品種黃萎病發生的相關性存在差異,感病品種黃萎病的發生程度與土壤中病原菌數量呈正相關。抗病品種在盛花期、開花后期和結鈴期土壤的細菌群落結構優于感病品種,并且不同生育時期的優勢菌群存在一定程度的差異。土壤中細菌多樣性、相對豐度和組成受有機質、pH、氮素類型、速效磷等指標影響。同時,棉花不同生育時期對土壤中細菌群落結構有明顯影響。

關鍵詞: 棉花黃萎病, 大麗輪枝菌, 抗病性, 花鈴期, 細菌群落結構, 高通量測序, 土壤理化性質

Abstract:

【Objective】The objective of this study is to research soil bacterial community structure of cotton verticillium wilt resistant and susceptible varieties at flowering and boll stage, understand the relationship between soil bacterial community structure and soil physicochemical properties, and to provide a theoretical basis for monitoring of cotton verticillium wilt and green ecological control. 【Method】The susceptible (EJ) and resistant (J863) varieties were used in field plot experiments. The ITS gene copy number of Verticillium dahliae and soil bacterial community structure at different flowering and boll stages (flowering, late flowering, and boll-forming stages) were studied by real-time fluorescence quantitative PCR (real-time PCR) and high throughput sequencing (Illumina MiSeq), respectively. The redundancy analysis (RDA) was used to determine the correlation between bacterial community structure and soil physicochemical properties. 【Result】There were different degrees of relevance between the occurrence of verticillium wilt and the ITS gene copy number of V. dahliae in soil. The incidence and disease index of susceptible variety EJ were positively correlated with the ITS gene copy number of V. dahliae, while those of resistant variety J863 were not significantly correlated with the ITS gene copy number of V. dahliae. The ITS gene copy number of V. dahliae in soil of resistant variety J863 was lower than that of susceptible variety EJ at late flowering and boll-forming stages. The high throughput sequencing analysis showed that the bacterial richness indices (Chao1 and ACE) of resistant variety J863 were higher than those of susceptible variety EJ at flowering and boll-forming stages. The principal component analysis showed that there were differences in soil bacterial community structure between resistant and susceptible varieties and at different flowering and boll stages. At phylum level, the average relative abundance of some dominant bacteria in susceptible variety EJ was lower than that of resistant variety J863, such as Actinobacteria, Gemmatimonadetes, Chloroflexi, Bacteroidetes, Nitrospirae, Patescibacteria, and Armatimonadetes, which was decreased by 16.38%, 4.05%, 2.25%, 6.58%, 7.10%, 20.60%, and 35.78%, respectively. At genus level, the average relative abundance of some dominant bacteria of susceptible variety EJ was lower than that of resistant variety J863, including Sphingomonas, Gemmatimonas, Bryobacter, Iamia, Pseudarthrobacter, Blastococcus, Rubrobacter, Nocardioides, Pontibacter, Streptomyces, Gemmatirosa, Micromonospora, and Solirubrobacter, which was decreased by 5.09%, 19.41%, 13.79%, 2.36%, 10.78%, 34.47%, 46.76%, 61.84%, 52.75%, 48.61%, 74.79%, 9.13%, and 26.42%, respectively. The redundancy analysis (RDA) showed that the bacterial community structure was affected by nitrate nitrogen (NO3 --N), available phosphorus (AP), ammonium nitrogen (NH4 +-N), inorganic phosphorus (IP), pH and organic matter (OM). 【Conclusion】The correlation between the ITS gene copy number of V. dahliae in soil and the occurrence of cotton verticillium wilt of susceptible and resistant varieties is different, and the incidence of verticillium wilt in susceptible variety is positively correlated with the ITS gene copy number of V. dahliae in soil. The bacterial community structure of resistant variety is better than susceptible variety at flowering, late flowering, and boll-forming stages. There are some differences in the dominant bacterial community at different growth stages. The diversity, relative abundance and composition of bacteria in soil are affected by organic matter, pH, nitrogen type and available phosphorus. Meanwhile, the growth stage of cotton had a significant influence on the bacterial community structure in soil.

Key words: cotton verticillium wilt, Verticillium dahliae, disease resistance, flowering and boll stage, bacterial community structure, Illumina MiSeq, soil physicochemical properties

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