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中国农学通报

中国农学通报 ›› 2023, Vol. 39 ›› Issue (9): 16-23.doi: 10.11924/j.issn.1000-6850.casb2022-0591

• 农学·农业基础科学 • 上一篇    下一篇

促生潜力玉米种子内生细菌分离鉴定及发酵条件优化

杨苔1,2,3(), 于慧颖1,2,3, 卢立明4, 付海燕1,2,3(), 李国良5, 代文龙1,2,3, 刘春光1,2,3, 杨峰山1,2,3, 马玉堃1,2,3()   

  1. 1 黑龙江大学农业微生物技术教育部工程研究中心，哈尔滨 150500
    2 黑龙江大学生命科学学院，黑龙江省寒地生态修复与资源利用重点实验室，哈尔滨 150080
    3 黑龙江大学生命科学学院，黑龙江省普通高校分子生物学重点实验室，哈尔滨 150080
    4 山东省莒县长岭镇农业综合服务中心，山东日照 276516
    5 黑龙江省农业科学院玉米研究所，哈尔滨 150080
  • 收稿日期:2022-07-21 修回日期:2022-10-27 出版日期:2023-03-25 发布日期:2023-03-23
  • 通讯作者: 付海燕，女，1977年出生，黑龙江哈尔滨人，副教授，博士，主要从事寒区有害生物生态防控研究。通信地址：150080 黑龙江省哈尔滨市学府路74号，黑龙江大学生命科学学院，Tel：0451-86609306，E-mail：fuhaiyan@hlju.edu.cn。马玉堃，男，1962年出生，辽宁沈阳人，教授，博士，主要从事生态学科研工作。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号 黑龙江大学生命科学学院，E-mail：mayukun@hlju.edu.cn。
  • 作者简介:

    杨苔，女，1997年出生，四川广元人，研究生，主要从事生态修复研究。通信地址：150080 黑龙江省哈尔滨市学府路74号 黑龙江大学生命科学学院，E-mail：。

  • 基金资助:
    国家自然科学基金“基于定量ITRAQ蛋白质组学解析MeJA诱导玉米抗玉米螟分子机制”(31672060); 黑龙江省自然科学基金研究团队项目“寒区农业土壤面源污染微生物调控技术的研究”(TD2019C002); 黑龙江省自然科学基金重点项目“茉莉酸甲酯诱导玉米抗玉米螟分子机制研究(ZD2017004); 黑龙江省蔬菜现代农业产业技术协同创新推广体系建设资金“蔬菜病虫害综合防控协同创新岗”(2021-2022)

Endophytic Bacteria from Maize Seed with Growth-promoting Potential: Isolation, Identification and Optimization of Fermentation Conditions

YANG Tai1,2,3(), YU Huiying1,2,3, LU Liming4, FU Haiyan1,2,3(), LI Guoliang5, DAI Wenlong1,2,3, LIU Chunguang1,2,3, YANG Fengshan1,2,3, MA Yukun1,2,3()   

  1. 1 Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500
    2 Heilongjiang Provincial Key Laboratory of Ecological Restoration and Resource Utilization for Cold Region, School of Life Sciences, Heilongjiang University, Harbin 150080
    3 Key Laboratory of Molecular Biology, College of Life Science, Heilongjiang University, Harbin 150080
    4 Changling Town Agricultural Comprehensive Service Center, Juxian County, Rizhao, Shandong 276516
    5 Maize Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150080
  • Received:2022-07-21 Revised:2022-10-27 Online:2023-03-25 Published:2023-03-23

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摘要：

旨在从玉米种子中分离鉴定具有促生功能的内生细菌，为菌肥研发提供菌种资源和技术支持。以玉米种子为试材，用平板涂布和三区划线法分离、纯化获得玉米种子内生菌，滴灌试验筛选促生菌株，采用单因素和正交试验优化发酵条件，通过盆栽滴灌试验验证菌株的促生效应。试验分离获得一株促生效果相对较强的菌株，编号为SYL4，初步鉴定为枯草芽孢杆菌(Bacillus subtilis)。SYL4最佳培养条件：蛋白胨1.5%，玉米粉0.5%，CaCO3 0.25%，接种量5%，转速200 r/min，温度34℃，培养时间48 h。滴灌优化发酵液使得玉米幼苗根长、叶长分别增加12.86%、9.4%，根重、叶重分别增加6.52%、6%。本研究分离获得一株具促生潜力的内生菌SYL4，优化后的发酵条件提高玉米幼苗过氧化物酶(POD)活性，有效促进玉米幼苗生长。

关键词: 玉米, 内生细菌, 酶活, 植物促生, 发酵条件优化

Abstract:

The aim of this study is to isolate and identify endophytic bacteria with growth-promoting function from maize seeds, and to provide strain resources and technical support for the research and development of bacterial fertilizer. In this experiment, maize seeds were used as the test materials, and endophytic bacteria were isolated and purified by plate coating and three-zone marking method. Drip irrigation test was used to screen growth-promoting strains. The fermentation conditions were optimized by single factor test and orthogonal test. The promoting effect of the strains was verified by pot drip irrigation test. Bacillus subtilis was isolated and identified as SYL4, which had a relatively strong growth-promoting effect. The optimal culture condition for SYL4 was: peptone 1.5%, corn flour 0.5%, CaCO3 0.25 %, inoculation volume 5%, rotation speed 200 r/min, temperature 34℃, and incubation time 48 h. Drip irrigation of maize with fermentation broth increased root length and leaf length by 12.86% and 9.4%, root weight and leaf weight by 6.52% and 6%, respectively. In this study, an endophytic bacterium SYL4 with growth-promoting potential was isolated. The optimized fermentation condition increased the activity of peroxidase (POD) in maize seedlings and effectively promoted the growth of maize seedlings.

Key words: maize, endophytic bacteria, enzyme activity, plant growth-promoting, optimization of fermentation conditions