Jha P N, Kumar A
Microbial Biotechnology Unit, School of Biotechnology, Banaras Hindu University, Varanasi 221 005, India.
J Appl Microbiol. 2007 Oct;103(4):1311-20. doi: 10.1111/j.1365-2672.2007.03383.x.
To isolate and characterize endophytic diazotrophic bacteria from a semi-aquatic grass (Typha australis) which grows luxuriantly with no addition of any nitrogen source.
Ten endophytic diazotrophic bacteria from surface-sterilized roots and culm of T. australis were isolated and screened for plant growth-promoting activities employing standard methods. Based on the rate of nitrogenase activity, indole acetic acid (IAA) production and phosphate (P) solubilization, one root isolate namely GR-3 was found to be the most efficient one. This isolate was identified as Klebsiella oxytoca on the basis of 16S rDNA sequence analysis. Amplification of nifH by polymerase chain reaction (PCR) and detection of dinitrogenase reductase by western blot confirmed the diazotrophic nature of GR-3. It was tagged with gusA fused to a constitutive promoter and the resulting transconjugant was inoculated onto endophyte-free rice variety Malviya dhan-36 seedlings to express cross-infection ability which resulted in a significant increase in root/shoot length and chlorophyll a content.
Roots and culm of T. australis harbour several endophytic diazotrophic bacteria. One root isolate, identified as K. oxytoca GR-3, seems to be an efficient plant growth-promoting bacterium.
Plant growth-promoting properties of GR-3 suggest that this promising isolate merits further investigations for potential application in agriculture.
从一种半水生草本植物(澳洲香蒲)中分离并鉴定内生固氮细菌,该植物在不添加任何氮源的情况下生长繁茂。
从表面消毒的澳洲香蒲根和茎中分离出10株内生固氮细菌,并采用标准方法筛选其促进植物生长的活性。根据固氮酶活性、吲哚乙酸(IAA)产量和磷(P)溶解率,发现一株根部分离菌GR-3是最有效的。基于16S rDNA序列分析,该分离菌被鉴定为产酸克雷伯菌。通过聚合酶链反应(PCR)扩增nifH并通过蛋白质免疫印迹检测二氮酶还原酶,证实了GR-3的固氮性质。用与组成型启动子融合的gusA对其进行标记,将所得的接合转移子接种到无内生菌的水稻品种Malviya dhan-36幼苗上,以表达交叉感染能力,这导致根/茎长度和叶绿素a含量显著增加。
澳洲香蒲的根和茎中含有几种内生固氮细菌。一种根部分离菌,鉴定为产酸克雷伯菌GR-3,似乎是一种有效的促进植物生长的细菌。
GR-3促进植物生长的特性表明,这种有前景的分离菌值得进一步研究,以探讨其在农业中的潜在应用。