芦竹属菌草内生固氮菌nifH基因的表达研究  

Expression of nif H Gene in Endophytic Nitrogen-fixing Bacteria of Arundo donax Lüzhou

作　　者：李曼 李巧琪 廖真 林标声 鲁国东[1,4] 林占熺[2] LI Man;LI Qiao-qi;LIAO Zhen;LIN Biao-sheng;LU Guo-dong;LIN Zhan-xi(School of Life Science,Fujian Agriculture and Forestry University,Fujian Fuzhou 350002,China;China National Engineering Research Center of Juncao Technology,Fujian Fuzhou 350002,China;College of Life Science,Longyan University,Fujian Longyan 364012,China;College of Plant Protection,Fujian Agriculture and Forestry University,Fujian Fuzhou 350002,China)

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]国家菌草工程技术研究中心,福建福州350002 [3]龙岩学院生命科学学院,福建龙岩364012 [4]福建农林大学植物保护学院,福建福州350002

出　　处：《西南农业学报》2021年第8期1776-1780,共5页Southwest China Journal of Agricultural Sciences

基　　金：中央引导地方科技发展专项资金项目“菌草固氮菌肥的开发及应用”(2018L003)。

摘　　要：【目的】检测5种芦竹属菌草绿洲成熟期根样本内生固氮菌的丰度及nifH基因的表达,为nifH基因的快速定量检测及实时监测提供技术手段。【方法】采用实时荧光定量PCR技术(Real-time quantitative reverse transcription PCR,qRT-PCR)分析根样本内生固氮菌nifH基因的拷贝数及相对表达量。【结果】5种芦竹属菌草成熟期根样本内生固氮菌nifH基因表达量及拷贝数均有较大的差异。其中,绿洲2号nifH基因相对表达量最高,呈过表达水平,且绿洲2号样本差异达显著水平(P<0.05),其余各样本差异不显著;固氮基因nifH的拷贝数呈逐步递减,其中绿洲1号样本nifH基因的拷贝数最高,达8.84×10^(11)/g;绿洲5号最低,达1.54×10^(11)/g,各样本的差异均达到显著水平。【结论】基于实时荧光定量PCR对固氮基因nifH的定量及定性分析,可为nifH基因的定量检测及表达情况提供理论依据。【Objective】The abundance and expression of nifH gene in root samples of five species of Arundo donax L.at the mature stage were detected,which provided a technical means for the rapid quantitative detection and real-time monitoring of nifH gene.【Method】Real-time quantitative reverse PCR(qRT-PCR)was used to analyze the copy number and relative expression of nifH gene in root samples.【Result】The nifH gene expression level and copy number of the root samples of five species of Arundo donax L.at the mature stage were significantly different.Among them,the relative expression level of nifH gene of Lüzhou 2 was the highest,showing an overexpression level,and the difference of Lüzhou 2 samples reached a significant level(P<0.05),the difference of other samples was not significant;The copy number of nifH of nitrogen fixation gene was gradually decreasing,and the copy number of nifH gene in Lüzhou 1 was the highest,reaching 8.84×10^(11)/g.Lüzhou 5 was the lowest,reaching 1.54×10^(11)/g,with significant differences among all samples.【Conclusion】Quantitative and qualitative analysis of nitrogen-fixing gene nifH based on real-time fluorescence quantitative PCR can provide theoretical basis for quantitative detection and expression of nifH gene.

关 键 词：芦竹属菌草 内生固氮菌 固氮基因 相对表达量 拷贝数 

分 类 号：S543.9[农业科学—作物学]