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[1]原红娟,严慧,杨芳,等.澳洲野生稻(Oryza australiensis)内生固氮菌的分子鉴定及发育分析[J].应用与环境生物学报,2014,20(04):571-577.[doi:10.3724/SP.J.1145.2013.11040]
 YUAN Hongjuan,YAN Hui,YANG Fang,et al.Molecular characterization and phylogenetic analysis of endophytic nitrogen-fixing bacteria in Oryza australiensis[J].Chinese Journal of Applied & Environmental Biology,2014,20(04):571-577.[doi:10.3724/SP.J.1145.2013.11040]
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澳洲野生稻(Oryza australiensis)内生固氮菌的分子鉴定及发育分析()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
20卷
期数:
2014年04期
页码:
571-577
栏目:
研究论文
出版日期:
2014-08-25

文章信息/Info

Title:
Molecular characterization and phylogenetic analysis of endophytic nitrogen-fixing bacteria in Oryza australiensis
作者:
原红娟 严慧 杨芳 彭桂香 谭志远
1华南农业大学农学院 广州 5106422运城学院生命科学系 运城 0440003华南农业大学资源环境学院 广州 510642
Author(s):
YUAN Hongjuan YAN Hui YANG Fang PENG Guixiang TAN Zhiyuan
1College of Agriculture, South China Agricultural University, Guangzhou 510642, China2Department of Life Science, Yuncheng University, Yuncheng 044000, China 3College of Resources Environment, South China Agricultural University, Guangzhou 510642, China
关键词:
澳洲野生稻遗传多样性内生固氮菌DNA指纹图谱系统发育分析
Keywords:
Oryza australiensis genetic diversity endophytic nitrogen-fixing bacteria DNA fingerprinting phylogenetic analysis
分类号:
Q939.11+3.03 : Q948.12+2.3
DOI:
10.3724/SP.J.1145.2013.11040
文献标志码:
A
摘要:
采用两种无氮培养基经平板划线法共分离筛选到澳洲野生稻(Oryza australiensis)45株内生固氮菌. 利用全细胞蛋白电泳和插入序列指纹图谱对获得的内生固氮菌进行聚类分析,将其分为8个类群. 其中类群Ⅰ有10株菌,类群Ⅱ为4株菌,类群Ⅲ、Ⅳ和Ⅴ均为3株菌,类群Ⅵ、Ⅶ和Ⅷ各有2株菌. 对分离得到的内生固氮菌主要类群部分代表菌株的固氮活性进行了系统研究,结果表明,菌株在偏酸(pH值5.0)和偏碱(pH值9.0)的条件下均能保持较高的固氮酶活性,NaCl浓度在2%时达到最高固氮活性,NH4+浓度达到7.5 mmol/L时,均无固氮酶活性,不同菌株所能利用的碳源不同. 16S rDNA序列测定及系统发育分析显示:类群ⅠYH39与Burkholderia cepacia ATCC 25416T相似性为97%;类群Ⅱ为克雷白氏杆菌属(Klebsiella),相似性为100%;类群Ⅲ为泛菌属(Pantoea),相似性为99%;类群Ⅴ为草螺菌属(Herbaspirillum),相似性为99%;类群Ⅶ为中华根瘤菌属(Sinorhizobium),相似性为99%. 本研究表明澳洲野生稻内生固氮菌资源具有遗传多样性. 图5 表1 参26
Abstract:
This research aimed to investigate the diversity of endophytic nitrogen-fixing bacteria in Oryza australiensis. Forty-five strains of endophytic diazotrophs were isolated from wild rice O. australiensis by using two selective nitrogen-free media, and analyzed for phylogenetic and nitrogen-fixing characteristics. The obtained endophytic nitrogen-fixing bacteria were divided into 8 clusters by using SDS-PAGE of whole-cell protein and IS-PCR fingerprinting. The strains were found to be able to maintain higher nitrogenase activity under acid (pH 5.0) or alkaline (pH 9.0) conditions. The highest nitrogen fixation activity occurred in 2% NaCl concentration. None of the strains showed nitrogenase activity at NH4+ of 7.5 mmol/L. Among the 8 clusters, Cluster Ⅰ had 10 strains, followed by 4 strains of Cluster Ⅱ, 3 of Clusters Ⅲ, Ⅳ and Ⅴ, and 2 of Clusters Ⅵ, Ⅶ and Ⅷ. The homology was 97% between Cluster Ⅰ and Burkholderia cepacia ATCC 25416T, 99% between Cluster Ⅴ and Herbaspirillum seropedicae DX35 T(KF357956.1), 100% between Cluster Ⅱ and Klebsiella variicola CPB S30T(KF770986.1), 99% between Cluster Ⅲ and Pantoea agglomerans PB17T(EU360112.1), and 99% between Cluster Ⅶ and Sinorhizobium sp. T2T(FJ687994.1). The results suggested that endophytic nitrogen-fixing bacteria of O. australiensis have genetic diversity, with different strains being able to use different carbon sources.

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备注/Memo

备注/Memo:
国家自然科学基金项目(31370052)、国家基础研究发展计划项目(2010CB126502)和广东省科技项目( 2013B060400020,2012B010300018)资助 Supported by the National Natural Science Foundation of China (NSFC) (31370052), the National Basic Research Program of China (973 Program) (2010CB126502), and the Science and Technology Program of Guangdong Province (2013B060400020, 2012B010300018)
更新日期/Last Update: 2014-08-26