健康与感染青枯病烟株根际土壤与茎秆细菌群落结构与多样性

Bacterial community structure and diversity of rhizosphere soil and stem of healthy and bacterial wilt tobacco plants

摘要:

[目的]了解健康烟株与感染青枯病烟株在根际土壤、茎杆发病部位、茎杆病健交界部位以及未发病茎杆的细菌群落结构与多样性。[方法]分别对土壤与茎杆样品中细菌的16S rRNA基因V3-V4区进行扩增，采用Illumina MiSeq测序技术对扩增片段进行高通量测序，然后对健康烟株与感染青枯病烟株不同部位细菌群落结构与多样性进行分析。[结果]感染青枯病烟株发病茎杆及根际土壤的细菌群落多样性高于健康烟株茎杆及其根际土壤样品，病健交界茎杆样品细菌群落多样性低于健康烟株。变形菌门（Proteobacteria）在所有样品中均为优势菌门；所有烟株根际土壤的优势菌门为拟杆菌门（Bacteroidetes）、酸杆菌门（Acidobacteria）、放线菌门（Actinobacteria）和绿弯菌门（Chloroflexi）；健康烟株茎杆部位的优势菌门为蓝细菌门（Cyanobacteria）；感染青枯病烟株发病茎杆和病健交界茎杆部位的优势菌门为蓝细菌门（Cyanobacteria）和厚壁菌门（Firmicutes）。所有根际土壤样品的优势菌属为劳尔氏菌属（Ralstonia）、假单胞菌属（Pseudomonas）、鞘脂单胞菌属（Sphingomonas）、黄杆菌属（Flavobacterium）和代尔夫特菌属（Delftia），而感染青枯病烟株根际土壤的劳尔氏菌属（Ralstonia）和假单胞菌属（Pseudomonas）相对丰度显著高于健康烟株根际土壤，鞘脂单胞菌属相对丰度显著低于健康烟株根际土壤。烟株茎杆的优势菌属为劳尔氏菌属和假单胞菌属等。感染青枯病烟株病健交界茎杆中劳尔氏菌属、肠杆菌属（Enterobacter）和泛菌属（Pantoea）相对丰度显著低于健康烟株样品。[结论]健康与感染青枯病烟株茎杆样品细菌群落的丰富度和多样性明显低于相应的根际土壤样品。较健康烟株而言，感染青枯病烟株根际土壤和茎杆样品细菌群落丰富度和多样性均表现出不同程度地增加，且根际土壤细菌群落结构变化较茎杆样品明显，而病健交界茎杆样品细菌群落丰富度和多样性降低。烟草青枯病为典型土传病害，其病原茄科劳尔氏菌尽管能在烟株维管束中蔓延扩增，但主要还是分布于土壤中；它的存在似乎对土壤细菌群落的影响大于茎杆样品的。该研究结果提示对于青枯病的防治不能局限于烟株本身，田间土壤也应加大防治力度。

Abstract:

[Objective] To understand the bacterial community structure and diversity of healthy and bacterial wilt tobacco plants in rhizosphere soil, diseased stem tissue, symptomatic-asymptomatic junction stem tissue, and non-infected stem tissue. [Methods] The V3-V4 region of the bacterial 16S rRNA gene in soil and stem tissue was amplified, and the amplified fragments were sequenced using Illumina MiSeq high-throughput sequencing technology. [Results] Proteobacteria were the dominant phylum in all samples. Bacteroidetes and Acidobacteria were the dominant phyla in all rhizosphere soil. Cyanobacteria was the dominant phylum in non-infected stem tissue; and Cyanobacteria and Firmicutes were dominant in diseased stem tissue and symptomatic-asymptomatic junction stemtissue. The dominant genera in all rhizosphere soil were Ralstonia, Pseudomonas and Sphingomonas. The dominant genera of tobacco stem were Ralstonia and Pseudomonas. [Conclusion] Compared with non-infected tobacco plants, the richness and diversity of bacterial community in rhizosphere soil and stem tissue of bacterial wilt tobacco plants were generally greater. Bacterial wilt is a typical soil-borne disease, although the pathogen Ralstonia solanacearum can spread and expand in the vascular bundles, it mainly distributes in the soil. Therefore, the control of bacterial wilt should not be limited to the tobacco plant itself, but targeting the pathogen in field soil may strengthen the control efforts.