不同抗、感白发病谷子品种内生细菌群落组成及差异性分析

doi:10.7668/hbnxb.20193151

华北农学报 ›› 2022, Vol. 37 ›› Issue (S1): 300-308. doi: 10.7668/hbnxb.20193151

所属专题： 杂粮作物；植物保护；

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不同抗、感白发病谷子品种内生细菌群落组成及差异性分析

邹晓悦¹^,², 刘佳¹, 董志平¹, 马继芳¹, 石爱丽³, 白辉¹, 李志勇¹

¹ 河北省农林科学院谷子研究所,农业农村部特色杂粮遗传改良与利用重点实验室(部省共建),河北省杂粮研究实验室, 河北石家庄 050035
² 河北师范大学生命科学学院,河北石家庄 050000
³ 承德市农林科学院,河北承德 067000

收稿日期:2022-09-28 出版日期:2022-12-28
作者简介:
邹晓悦(1995-),女,河北张家口人,在读硕士,主要从事谷子抗病分子生物学研究。邹晓悦、刘佳为同等贡献作者。
基金资助:
国家重点研发计划(2018YFD1000703); 国家重点研发计划(2018YFD1000700); 河北省农林科学院基本业务费(2018030202); 河北省农林科学院基本业务费(2021030201); 河北省农林科学院基本科研业务费包干制项目资助(HBNKY-BGZ-02); 国家现代农业产业技术体系(CARS-06-14.5-A25); 河北省重点研发计划(21326338D)

The Composition and Difference Analysis of Endophytic Bacterial Community in Different Resistant/Susceptible Foxtail Millet Varieties

ZOU Xiaoyue¹^,², LIU Jia¹, DONG Zhiping¹, MA Jifang¹, SHI Aili³, BAI Hui¹, LI Zhiyong¹

¹ Institute of Millet Crops,Hebei Academy of Agriculture and Forestry Sciences,Key Laboratory of Genetic Improvement and Utilization for Featured Coarse Cereals Coconstruction by Ministry and Province, Minist of Agriculture and Rur Affairs,Key Laborat of Minor Cereal Crops of Hebei Province,National Foxtail Millet Improvement Center,Shijiazhuang 050035,China
² College of Life Sciences, Hebei Normal University,Shijiazhuang 050000,China
³ Chengde Academy of Agriculture and Forestry Sciences,Chengde 067000,China

Received:2022-09-28 Published:2022-12-28

摘要/Abstract

摘要：

明确不同品种谷子内生细菌群落组成特征,揭示与白发病抗性相关的关键物种,为利用内生细菌防治谷子白发病提供理论依据。对3种抗白发病和3种感白发病谷子植株的第一节茎与最高节茎组织进行DNA提取和样品中细菌16S rDNA高通量测序,并对内生细菌的群落组成进行分析。结果发现,在各分类水平下,从第一节茎到最高节茎内生细菌种类在抗病、感病谷子样品中分别表现为基本稳定与明显下降的2种不同趋势,且抗病谷子内生细菌群落的物种组成均较感病谷子丰富。得到重要内生细菌门19种:对普遍存在于抗病、感病谷子样品第一节茎中的内生细菌进行比较得到抗病谷子特有内生细菌门2种;对抗病、感病谷子样品各自全部内生细菌进行比较得到抗病谷子特有内生细菌门16种以及抗病谷子相对丰度较大的优势类群2种。可见,不同谷子品种以及不同部位内生细菌群落组成具有差异,抗病较感病谷子内生细菌种类丰富,其中抗病谷子特有的内生细菌门以及相对丰度大的优势细菌门对谷子白发病可能具有生物防治作用,值得进一步研究。

关键词: 谷子, 白发病, 内生细菌, Miseq高通量测序

Abstract:

In order to clarify the characteristics of the endophytic bacterial community of different varieties of foxtail millet,reveal the key species related to foxtail millet downy mildew resistance.DNA extraction was performed on the lowest node stem and the highest node stem tissues of 3 kinds of foxtail millet downy mildew-resistant and 3 kinds of foxtail millet downy mildew-susceptible foxtail millet plants.We used Illumina Miseq to sequence the bacterial 16S rDNA of the samples and analyzed the susceptible community composition of endophytic bacteria.The results showed that at each classification level,the endophytic bacterial species from the lowest node stem to the highest node stem showed two different trends in disease-resistant and susceptible foxtail millet samples,which were basically stable and significantly decreased.And the species composition of the endophytic bacterial community in disease-resistant foxtail millet was more abundant than that of susceptible foxtail millet.19 important endophytic bacteria phyla were obtained:(from the comparison of the endophytic bacteria ubiquitous in the lowest node stem of disease-resistant and susceptible foxtail millet samples were obtained) 2 species of endophytic bacteria phyla specific to disease-resistant foxtail millet;(from the comparison of all endophytic bacteria of disease-resistant and susceptible foxtail millet samples 16 species of endophytic bacteria phyla unique to disease-resistant foxtail millet and 2 dominant groups with relatively large abundance of disease-resistant foxtail millet were obtained).It can be seen that the endophytic bacterial communities in different foxtail millet varieties and different parts are different,and the endophytic bacterial species of disease resistance are more abundant than that of susceptible foxtail millet.Among them,the endophytic bacteria phyla unique to disease-resistant foxtail millet materials and the dominant endophytic bacteria phyla with relatively large abundance may have an effect on the prevention and treatment of foxtail millet downy mildew,which is worthy of further study.

Key words: Foxtail millet, Downy mildew, Endophytic bacteria, Miseq high-throughput sequencing

邹晓悦, 刘佳, 董志平, 马继芳, 石爱丽, 白辉, 李志勇. 不同抗、感白发病谷子品种内生细菌群落组成及差异性分析[J]. 华北农学报, 2022, 37(S1): 300-308. doi: 10.7668/hbnxb.20193151.

ZOU Xiaoyue, LIU Jia, DONG Zhiping, MA Jifang, SHI Aili, BAI Hui, LI Zhiyong. The Composition and Difference Analysis of Endophytic Bacterial Community in Different Resistant/Susceptible Foxtail Millet Varieties[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(S1): 300-308. doi: 10.7668/hbnxb.20193151.

http://www.hbnxb.net/CN/Y2022/V37/IS1/300

图/表 7

表1 扩增引物序列

Tab.1 Amplification primer sequence

巢式PCR Nested PCR	引物 Primer	序列(5'-3') Sequence(5'-3')	扩增区域 Target region
第1轮	799F	AACMGGATTAGATACCCKG	16S rDNA
First round	1392R	ACGGGCGGTGTGTRC	799F-1392R
第2轮	799F	AACMGGATTAGATACCCKG	16S rDNA
Second round	1193R	ACGTCATCCCCACCTTCC	799F-1193R

图1 谷子样品Coverage指数图 R.抗病;S.感病;LS.第一节茎;HS.最高节茎;1,2,3.样品顺序号。

Fig.1 Coverage index of foxtail millet samples R.Disease-resistant;S.Disease-susceptible;LS.The lowest node stem;HS.The highest node stem;1,2,3.The material number.

图2 谷子样品Shannon稀释性曲线图

Fig.2 Shannon dilution curve of foxtail millet samples

表2 不同分类水平下抗、感病谷子样品内生细菌种类

Tab.2 Species of endophytic bacteria in resistant and susceptible samples of foxtail millet at different classification levels

分类水平 Classification level	门 Phylum	纲 Class	目 Order	科 Family	属 Genus	种 Species
R_LS	23	40	97	224	423	647
R_HS	27	47	106	218	421	606
S_LS	19	35	90	202	345	513
S_HS	11	17	48	94	163	238

图3 谷子样品第一节茎内生细菌门比较 A.抗病谷子样品第一节茎内生细菌门分布韦恩图;B.感病谷子样品第一节茎内生细菌门分布韦恩图;C.抗病谷子与感病谷子样品第一节茎内生细菌门分布韦恩图;D.抗病谷子与感病谷子样品第一节茎优势内生细菌门分布柱形图。R1.山西红谷;R2.长农46号;R3.郑17-3415;S1.郑17-2123;S2.K148-1;S3.保谷25号;R.3份抗病谷子;S.3份感病谷子;LS.第一节茎。图4-5同。

Fig.3 Comparison of endophytic bacteria phylum in the lowest node stem of foxtail millet samples A.Venn diagram of distribution of endophytic bacteria phyla in the lowest node stems of disease-resistant foxtail millet samples;B.Venn diagram of the distribution of endophytic bacterial phyla in the lowest node stems of disease-susceptible foxtail millet samples;C.Venn diagram of distribution of endophytic bacterial phyla in the lowest node stems of disease-resistant and susceptible foxtail millet samples;D.Histogram of distribution of dominant endophytic bacteria phyla in the lowest node stems of disease-resistant and susceptible foxtail millet samples.R1.Shanxihonggu;R2.Changnong 46;R3.Zheng 17-3415;S1.Zheng 17-2123;S2.K148-1;S3.Baogu 25;R.3 resistant samples of foxtail millet;S.3 susceptible samples of foxtail millet;LS.The lowest node stem.The same as Fig.4-5.

图4 谷子样品最高节茎内生细菌门比较 A.抗病谷子样品最高节茎内生细菌门分布韦恩图;B.感病谷子样品最高节茎内生细菌门分布韦恩图;C.抗病谷子与感病谷子样品最高节茎内生细菌门分布韦恩图;D.抗病谷子与感病谷子样品最高节茎优势内生细菌门分布柱形图。HS.最高节茎。图5同。

Fig.4 Comparison of endophytic bacteria phylum in the highest node stem of foxtail millet samples A.Venn diagram of distribution of endophytic bacteria phyla in the highest node stems of disease-resistant foxtail millet samples;B.Venn diagram of the distribution of endophytic bacterial phyla in the highest node stems of disease-susceptible foxtail millet samples;C.Venn diagram of distribution of endophytic bacterial phyla in the highest node stems of disease-resistant and susceptible foxtail millet samples;D.Histogram of distribution of dominant endophytic bacteria phyla in the highest node stems of disease-resistant and susceptible foxtail millet samples.HS.The highest node stem.The same as Fig.5.

图5 谷子样品不同部位内生细菌门比较 A.抗病谷子样品第一节茎与最高节茎内生细菌门分布韦恩图;B.感病谷子样品第一节茎与最高节茎内生细菌门分布韦恩图;C.抗病谷子与感病谷子样品第一节茎与最高节茎优势内生细菌门分布柱形图。

Fig.5 Comparison of endophytic bacteria phylum in different parts of foxtail millet samples A.Venn diagram of the distribution of endophytic bacterial phyla in the lowest node stems and highest node stems of disease-resistant foxtail millet samples;B.Venn diagram of the distribution of endophytic bacterial phyla in the lowest node stems and highest node stems of disease-susceptible foxtail millet samples;C.Histogram of the distribution of dominant endophytic bacteria phyla in the lowest node stems and highest node stems of disease-resistant and susceptible foxtail millet samples.

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不同抗、感白发病谷子品种内生细菌群落组成及差异性分析

The Composition and Difference Analysis of Endophytic Bacterial Community in Different Resistant/Susceptible Foxtail Millet Varieties

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不同抗、感白发病谷子品种内生细菌群落组成及差异性分析

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