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作物杂志,2024, 第4期: 194–202 doi: 10.16035/j.issn.1001-7283.2024.04.025

• 生理生化·植物营养·栽培耕作 • 上一篇    下一篇

酿酒专用糯红高粱内生细菌多样性分析及功能菌株筛选

胡连清1,2(), 陈露1,2, 刘雯雯1,2, 周万海1,2(), 冯瑞章1,2, 魏琴1,2, 赵鑫1,2, 舒豪1, 陈玲妹1, 陈雨薇3   

  1. 1宜宾学院农林与食品工程学部，644000，四川宜宾
    2香料植物资源开发与利用四川省高校重点实验室，644000，四川宜宾
    3简阳市农业技术推广中心，641400，四川简阳
  • 收稿日期:2023-06-26 修回日期:2023-11-10 出版日期:2024-08-15 发布日期:2024-08-14
  • 通讯作者: 周万海，主要从事微生物多样性与功能鉴定等工作，E-mail：wanhaizhou@126.com
  • 作者简介:胡连清，主要从事农业资源利用与开发等工作，E-mail：120726584@qq.com
  • 基金资助:
    四川省自然科学基金(2022NSFSC0176);四川省科技厅项目(2018JY0495);四川省油樟工程技术研究中心2021年度开放基金(21YZY06)

Investigation on the Diversity of Endophytic Bacteria and Screening of Functional Strains in Waxy Sorghum for Brewing

Hu Lianqing1,2(), Chen Lu1,2, Liu Wenwen1,2, Zhou Wanhai1,2(), Feng Ruizhang1,2, Wei Qin1,2, Zhao Xin1,2, Shu Hao1, Chen Lingmei1, Chen Yuwei3   

  1. 1Faculty of Agriculture, Forestry and Food Engineering, Yibin University, Yibin 644000, Sichuan, China
    2Key Laboratory of Aromatic Plant Resources Exploitation and Utilization in Sichuan Higher Education, Yibin 644000, Sichuan, China
    3Jianyang Agricultural Technology Extension Center, Jianyang 641400, Sichuan, China
  • Received:2023-06-26 Revised:2023-11-10 Online:2024-08-15 Published:2024-08-14

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摘要：

为分析糯红高粱不同品种、不同组织及不同物候期内生细菌群落结构及功能特点，利用纯培养法对四川宜宾地区酿酒专用糯红高粱内生细菌多样性及群落结构进行探究与分析，并通过选择培养基筛选其中的功能菌。结果表明，从杂交品种金糯粱1号和地方品种青壳洋中分离得到183株内生细菌，隶属于2门5纲17属39种，优势门为变形菌门（Proteobacteria），占85.79%，优势纲为γ-变形菌纲，占70.49%，优势属为假单胞菌属（Pseudomonas），占43.72%，优势种为Pseudomonas sp.，占33.33%。由此可知，糯红高粱内生细菌种类多样、功能丰富，不同品种的糯红高粱以及糯红高粱的不同组织、不同生育期独有内生细菌种群组成存在较大的差异。

关键词: 酿酒高粱, 糯红高粱, 内生细菌, 群落结构, 多样性

Abstract:

The diversity and community structure of endophytic bacteria in waxy sorghum for brewing were examined and analyzed using pure culture in order to examine the functional traits and community structure of endophytic bacteria of various varieties and tissues from waxy sorghum for brewing during different phenological periods and the choice of culture medium was used to filter the functioning bacteria. The results showed that 183 bacteria were isolated from the hybrid Jinnuoliang 1 and the local variety Qingkeyang, belonging to two phyla, five classes and 17 genera and 39 species. The dominant phyla was Proteobacteria, accounting for 85.79%, and the dominant class was γ-Protea, accounting for 70.49%, the dominant genus was Pseudomonas, accounting for 43.72%, and the dominant species was Pseudomonas sp., accounting for 33.33%. Therefore, endophytic bacteria from waxy sorghum were diverse in varieties and rich in functions. These research indicated that there are significant differences in the composition of unique endophytic bacterial populations among different varieties and tissues from waxy sorghum for brewing during different phenological periods.

Key words: Brewing sorghum, Waxy sorghum, Endophytic bacteria, Community structure, Diversity

表1

糯红高粱各组织中内生细菌定殖率及分离率

植物组织Plant tissue 品种Variety 生育期Growth stage 定殖率Colonization rate 分离率Separation rate
根Root 金糯粱1号 苗期、花期、蜡熟期 25.33 21.33
青壳洋 苗期、花期、蜡熟期 33.33 37.33
茎Stem 金糯粱1号 苗期、花期、蜡熟期 8.00 26.67
青壳洋 苗期、花期、蜡熟期 18.67 24.00
叶Leaf 金糯粱1号 苗期、花期、蜡熟期 4.00 5.33
青壳洋 苗期、花期、蜡熟期 3.33 2.00
花Flower 金糯粱1号 花期 6.00 6.00
青壳洋 花期 6.00 10.00

表2

糯红高粱内生细菌组成

相似性最高菌株
The most similar strain		 金糯梁1号Jinnuoliang 1 青壳洋Qingkeyang
苗期
Seedling stage		 花期
Florescence stage		 蜡熟期
Waxy stage		 苗期
Seedling stage		 花期
Florescence stage		 蜡熟期
Waxy stage
根
Root		 茎
Stem		 叶
Leaf		 根
Root		 茎
Stem		 叶
Leaf		 花
Flower		 根
Root		 茎
Stem		 叶
Leaf		 根
Root		 茎
Stem		 叶
Leaf		 根
Root		 茎
Stem		 叶
Leaf		 花
Flower		 根
Root		 茎
Stem		 叶
Leaf
醋酸钙不动杆菌
Acinetobacter calcoaceticus		 - - - - - - - - - - - - - 1 - - - - - -
不动杆菌Acinetobacter sp. - 1 - 1 - - - - - - - - - - - - - - - -
活不动杆菌Acinetobacter vivianii - - - - - - - - - - - - - 4 - - - - - -
堀越氏芽胞杆菌Bacillus horneckiae - - - - - - - - - - - - - - - - - 1 - -
同温层芽孢杆菌Bacillus stratosphericus - - - - - - - - - - - - - - 1 - - - - -
Bacterium - - - - - - - - - - - - - - - - - - 1 1
Bakterija - - - 1 1 - - - - - - - - 1 - - - - - -
唐菖蒲伯克霍尔德氏菌
Burkholderia gladioli		 - - - - - - - - 1 - - - - - - - - - 2 -
伯克霍尔德氏菌Burkholderia sp. - - - - - - - 1 - - - - - - - - - - 1 1
热带伯克霍尔德氏菌
Burkholderia tropica		 - - - - - - - - 1 - - - - - - - - - 1 -
柠檬酸杆菌Citrobacter sp. - - - - - - - 2 - - - - - - - - - - - 1
贪铜杆菌Cupriavidus sp. - - - - - - - - - - - - - 1 - - - - - -
阿氏肠杆菌Enterobacter asburiae - - - - - - - 1 - - - - - - - - - - - -
阴沟肠杆菌Enterobacter cloacae - - - - - - - - - - - - - - - - - 1 - -
霍氏肠杆菌Enterobacter hormaechei - - - - - - - - - - - - - 1 - - - 1 2 -
科贝肠杆菌Enterobacter kobei - - - - - - - - 1 - - - - - - - - - 1 -
罗根肠杆菌
Enterobacter roggenkampii		 - - - - - - - 3 - - - - - - - - - 10 7 -
肠棒杆菌Enterobacter sp. - - - - - - - 1 1 1 - - - 4 - - - - 4 -
香坊肠杆菌
Enterobacter xiangfangensis		 - - - - - - - - - - - - - - - - - - 1 -
γ蛋白细菌
Gamma proteobacterium symbiont		 - - - - - - - - - - - - - - 1 - - - - -
内生固氮菌
Herbaspirillum frisingense		 - - - - - - - - - - - - - - 1 - - - - -
哈特草螺菌Herbaspirillum huttiense - - - - - - - - - - - - - 3 - - 1 - - -
乌头内生菌Herbaspirillum sp. - - - - - - - - - - - - - 2 - - - - - -
Hrizaobakterijum sp. - - - - 2 - - - - - - - - - - - - - - -
肺炎克雷伯菌Klebsiella pneumoniae - - - - - - - - - - - - - - - - - - 1 -
克雷伯氏菌Klebsiella sp. - - - - - - - - - - - - - - 1 - - - - -
Komamonas aquatica - - - 1 - - - - - - - - - - - - - - - -
Metilobakterijum sp. - - - - - - 2 - - - - - - - - - - - - -
Nekulturna bakterija - - - 2 - - 1 - - - - - - - - - - - - -
Nekulturna beta proteobakterijum - - - - - - - - - - - - - 3 - - 2 - - -
Nekulturni Citrobacter sp. - - - 1 - - - - - - - - - 2 - - - - - -
叶杆菌
Phyllobacterium brassicacearum		 - - - 1 - - - - - - - - - - - - - - - -
绿针假单胞菌
Pseudomonas chlororaphis		 - - - - 1 - - - - - - - - - - - - - - -
荧光假单胞菌
Pseudomonas fluorescens		 - - - - - - - 1 - - - - - - - - - - - -
湖南假单胞菌
Pseudomonas hunanensis		 - - - - - - - 1 - - - - - - - - - - - -
浅黄色假单胞菌Pseudomonas lurida - - - - 3 - - - - - - - - - - - - - - -
蒙氏假单胞菌Pseudomonas monteilii - - - - 1 - - 2 - - - - - - - - - - - -
变形假单胞菌
Pseudomonas plecoglossicida		 - - - - - - - 2 - - - - - - - - - - - -
恶臭假单胞菌Pseudomonas putida - - 1 - 4 - - - - - - 1 - - - - - 1 1 -
假单胞菌Pseudomonas sp. - 12 3 6 10 1 - 3 - - 3 2 - 13 3 - 2 1 - -
葡萄牙根瘤菌Rhizobium lusitanum 2 - - - - - - - - - - - - 1 - - - - - -
根瘤菌Rhizobium sp. - - - - - - - - - - - - - - - - - 1 - -
鞘氨醇单胞菌Sphingomonas echinoides - - 2 - - - - - - - - - - - - - - - - -
Uncultured bacterium - 1 - - - - - - - - - 2 - - - - - 1 2 -

表3

糯红高粱不同品种、不同组织以及不同生育期内生细菌的丰富度指数、香农指数与优势度指数

项目
Item		 品种Variety 组织Tissue 生育期Growth stage
金糯粱1号
Jinnuoliang 1		 青壳洋
Qingkeyang		 根
Root		 茎
Stem		 叶
Leaf		 花
Flower		 苗期
Seedling stage		 花期
Florescence stage		 蜡熟期
Waxy stage
丰富度指数Abundance degree (S) 23 27 36 21 6 6 5 22 18
香农指数Shannon-Wiener index (H) 2.2249 2.6459 2.5629 2.3943 1.5811 1.7481 0.9321 2.4410 2.5648
优势度指数Dominance degree (D) 0.7529 0.8822 0.8590 0.8386 0.7407 0.8200 0.4400 0.8160 0.8728

图1

糯红高粱不同品种、不同组织以及不同生育期内生细菌的相似性系数

图2

不同品种、不同组织以及不同时期OTU数量的Venn图

表4

糯红高粱功能内生细菌筛选

菌株
Bacterial strain		 产淀粉酶
Amylase
production		 产蛋白酶
Protease
production		 产脂肪酶
Lipase
production		 产果胶酶
Pectinase
production		 产纤维素酶
Cellulase
production		 产酯酶
Esterase
production		 固氮能力
Nitrogen
fixation		 溶磷能力
Phosphate
solubilization
不动杆菌Acinetobacter sp. - - + - - - - -
活不动杆菌Acinetobacter vivianii - - + - - - - -
唐菖蒲伯克霍尔德氏菌Burkholderia gladioli + - + - + - - -
伯克霍尔德氏菌Burkholderia sp. - - - - - + - -
热带伯克霍尔德氏菌Burkholderia tropica - - + - - + - -
柠檬酸杆菌Citrobacter sp. - - - - - - - +
贪铜杆菌Cupriavidus sp. + - + - - - - -
阿氏肠杆菌Enterobacter asburiae - - - - - - + -
科贝肠杆菌Enterobacter kobei + - - - - - - -
罗根肠杆菌Enterobacter roggenkampii - + - + - + + -
肠棒杆菌Enterobacter sp. + - - - - - - -
香坊肠杆菌Enterobacter xiangfangensis - - + - - + - -
肺炎克雷伯菌Klebsiella pneumoniae - - - - - - - +
Komamonas aquatica + - - - - - - -
Metilobakterijum sp. + - + - - - - -
浅黄色假单胞菌Pseudomonas lurida + - + - - + - -
蒙氏假单胞菌Pseudomonas mosselii - + - + - - - -
恶臭假单胞菌Pseudomonas putida - + + + - - - -
假单胞菌Pseudomonas sp. + + - + - + + -
根瘤菌Rhizobium sp. - - - - - - + -
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