抗、感青枯病桑树根际细菌群落结构与多样性

武华周; 娄德钊; 涂娜娜; 耿涛; 卢芙萍; 吉训聪; 王树昌

doi:10.19303/j.issn.1008-0384.2020.09.012

抗、感青枯病桑树根际细菌群落结构与多样性

doi: 10.19303/j.issn.1008-0384.2020.09.012

武华周^1,,
娄德钊^{1, 2},
涂娜娜^{1, 2},
耿涛¹,
卢芙萍¹,
吉训聪^{3, 4},
王树昌^1, ,

1.
中国热带农业科学院环境与植物保护研究所，海南　海口　571101
2.
海南大学，海南海口 570228
3.
海南省农业科学院植物保护研究所海南　海口　571100
4.
海南省植物病虫害防控重点实验室，海南　海口　571100

基金项目: 国家现代农业产业技术体系专项（CARS-18-SYZ17）；中国热带农业科学院基本科研业务费专项（1630042019021）；中央引导地方科技发展专项（ZY2018HN01）

详细信息

作者简介:
武华周（1987−），男，硕士，助理研究员，研究方向：桑树栽培与育种（E-mail：wuhuazhouaa@126.com）

通讯作者:
王树昌（1976−），男，博士，研究员，研究方向：热带桑蚕技术研究（E-mail：wangshuchang2001@163.com）

中图分类号: S 432
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出版历程
- 收稿日期: 2020-08-10
- 修回日期: 2020-09-08
- 刊出日期: 2020-09-28

Rhizosphere Bacterial Community and Diversity at Fields of Wilt Resistant or Susceptible Mulberry Trees

WU Huazhou^1
,,
LOU Dezhao^{1, 2},
TU Nana^{1, 2},
GENG Tao¹,
LU Fuping¹,
JI Xuncong^{3, 4},
WANG Shuchang^{1
, ,}

1.
Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan　571101, China
2.
Hainan University, Haikou, Hainan　570228, China
3.
Institute of Plant Protection, Hainan Academy of Agricultural Sciences, Haikou, Hainan　571100, China
4.
Key Laboratory of Plant Disease and Pest Control of Hainan Province, Haikou, Hainan　571100, China

摘要

摘要: 目的通过抗、感青枯病桑树根际土壤细菌群落结构与多样性分析，阐明根际细菌群落与桑树抗、感青枯病的相关性。方法采用Illumina MiSeq测序技术对抗青枯病桑树基因型（抗青283×抗青10）和感病基因型（桂桑优62）根际细菌的16S rRNA基因V3-V4区进行扩增，高通量测序后比较分析了其根际细菌群落结构与多样性。结果 ① 抗、感青枯病桑树根际土壤的优势菌门为变形菌门、放线菌门、酸杆菌门、芽单胞菌门和拟杆菌门，优势菌属为MND1、Gaiella、硝化螺菌属、Haliangium和链霉菌属；② 抗、感青枯病桑树根际土壤细菌群落Alpha多样性无显著差异，但经NMDS排序，则可显著区分（stress=0.005＜0.05），其中抗性基因型根际土壤中具有重要作用的细菌类群较多，包括：硝化螺旋菌纲、酸杆菌纲、硝化螺旋菌目、索利氏菌目、酸杆菌目、硝化螺旋菌科、酸杆菌目未培养科、索利氏菌科Subgroup 3和Subgroup 2未培养科，而敏感基因型桂桑优62根际重要作用的细菌类群则主要为Ilumatobacteraceae和TRA3 20 Other两个科。③ 抗、感青枯病桑树根际细菌差异COG类目为763，仅占17.25%。结论抗、感青枯病桑树根际土壤细菌群落丰富度和多样性虽无显著差异，但其重要作用的类群显著不同，说明桑树根际土壤细菌群落结构可能与种质对青枯病的抗性具有相关性。本结果将为进一步研究桑树根际微生态特征、筛选有益功能菌株并用于桑青枯病的有效防控具有重要意义。
- 桑树 /
- 青枯病 /
- Illumina高通量测序 /
- 细菌多样性 /
- 群落结构
Abstract: Objective Structure and diversity of rhizosphere bacterial communities at fields of mulberry trees resistant (QZ2K) or susceptible (QZ2G) to wilt disease were studied. Method The V3-V4 regions of 16S rRNA in rhizosphere bacteria were amplified and sequenced using high-throughput sequencing technology on Illumina MiSeq to determine the bacterial community structure, diversity, and functions. Results from the two field samples were compared. Result (1) At phylum and genus levels, the dominant rhizosphere bacteria were similar at QZ2K (Kangqing 283×Kangqing 10 mulberry field) and at QZ2G (Guisangyou 62 mulberry field). The phyla included Proteobacteria, Actinobacteria, Acidobacteria, Gemmatimonadetes, and Bacteroidetes, while the genera consisted of MND1, Gaiella, Nitrospira, Haliangium, and Streptomyces. (2) Although no significant difference in the alpha diversity of the bacteria communities at the two different fields, the NMDS ordination showed significant differences (stress 0.005<0.05). At QZ2K, the bacteria related to significant metabolic functions were Nitrospira, Acidobacteriia, Nitrospirales, Solibacteraies, Acidobacteriales, Nitrospiraceae, and the uncultured Acidobacteria, Solibacteraies Subgroup 3, and Solibacteraies Subgroup 2. At QZ2G, only Ilumatobacteraceae and TRA3-20-other were identified. (3) According to the Wilcoxon signed rank test, the 763 different rhizosphere bacteria orthologs identified were only 17.25% of all COG orthologs on both fields. Conclusion There were no significant differences in the richness and diversity of rhizosphere bacteria community between the two fields. However, the bacteria associated with significant functions differed significantly between them, which could well be the species that made the difference in the occurrence of wilt disease on the mulberry plants. The information obtained in this study was of value for further studies on the microecological characteristics of mulberry rhizosphere as well as selection and application of functional bacteria for wilt control.
- Mulberry /
- bacterial wilt /
- Illumina high-throughput sequencing /
- bacterial diversity /
- community structure

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图 1 抗感青枯病桑树根际土壤细菌OTUs韦恩图

Figure 1. Venn diagram of bacteria OTUs in rhizosphere soils at QZ2K and QZ2G

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图 2 细菌群落门（A）和属（B）水平相对丰度

Figure 2. Phylum （A） and genus （B） distributions of OTUs

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图 3 抗感青枯病桑树根际土壤细菌NMDS分析

Figure 3. Non-MetricMulti-dimensional scaling by UUF metric

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图 4 抗感青枯病桑树根际土壤细菌LEfSe分析

Figure 4. LEfSe analysis on bacteria in rhizosphere soils at QZ2K and QZ2G

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图 5 抗感青枯病桑树根际细菌预测得到的COG相对丰度

注：A: RNA加工与修饰; B: 染色体结构与动态；C: 能量产生与转化; D: 细胞周期控制，细胞分裂，染色体分裂; E: 氨基酸转运与代谢; F: 核酸转运与代谢;G:碳水化合物的运输与代谢; H: 酶运输与代谢; I: 脂质运输与代谢; J:翻译，核糖体结构和生物发生; K: 转录; L: 复制，重组和修复; M: 细胞壁/膜/包膜生物发生; N: 细胞运动; O: 翻译后修饰，蛋白质更新，伴侣; P: 无机离子的运输与代谢; Q:次生代谢产物的生物合成，转运和分解代谢; R:仅通用功能预测; S:未知功能; T:信号转导机制; U:细胞内运输，分泌和囊泡运输; V: 防御机制; W: 细胞外结构; Z: 细胞骨架。

Figure 5. Relative abundance of COG at QZ2K and QZ2G

Note: A: RNA processing and modification; B: Chromatin structure and dynamics; C: Energy production and conversion; D: Cell cycle control, cell division, and chromosome partitioning; E: Amino acid transport and metabolism; F: Nucleotide transport and metabolism; G: Carbohydrate transport and metabolism; H: Coenzyme transport and metabolism; I: Lipid transport and metabolism; J: Translation, ribosomal structure and biogenesis; K: Transcription; L: Replication, recombination, and repair; M: Cell wall/membrane/envelope biogenesis; N: Cell motility; O: Posttranslational modification, protein turnover, and chaperones; P: Inorganic ion transport and metabolism; Q: Secondary metabolites biosynthesis, transport, and catabolism; R: General function prediction only; S: Function unknown; T: Signal transduction mechanisms; U: Intracellular trafficking, secretion, and vesicular transport; V: Defense mechanisms; W: Extracellular structures; Z: Cytoskeleton.

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表 1 高通量测序数据概况

Table 1. Overview of high-throughput sequencing data

样品 Sample	高质量序列 Clean tags	优质序列 Valid tags	平均长度 /bp Valid mean length	测序深度指数/% Goods coverage
QZ2K.1	33 883	28 948	429	94.81
QZ2K.2	38 786	32 371	430	94.69
QZ2K.3	32 328	26 941	432	94.57
QZ2K.4	41 499	34 267	432	94.56
平均值 average	36 624.0	30 631.8	430.8	94.658
QZ2G.1	38 747	32 496	432	94.53
QZ2G.2	37 626	31 688	432	94.43
QZ2G.3	40 749	34 814	432	94.78
QZ2G.4	40 656	34 090	432	95.22
平均值 average	3 944.5	33 272.0	432.0	94.740

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表 2 抗感青枯病桑树根际土壤细菌α多样性指数（平均值±标准误，n=4）

Table 2. Alpha diversity indices of bacteria in rhizosphere soils at QZ2K and QZ2G（Mean±SE，n=4）

样品 Samples	谱系多样性指数 PD whole Tree	Chao1	文库覆盖率（%） Goods coverage	观测物种数 Observed Species	香浓-威纳指数 Shannon-Wiener	辛普森指数 Simpson
QZ2K	84.97±0.77	3 190.57±22.48	0.95±0.00	2 333.88±40.49	9.51±0.09	0.995±0.001
QZ2G	82.86±1.44	3 178.61±87.45	0.95±0.00	2 304.30±49.58	9.65±0.03	0.997±0.001

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