National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box 577, Faisalabad, Pakistan.
Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.
Appl Microbiol Biotechnol. 2018 Jan;102(1):485-497. doi: 10.1007/s00253-017-8609-6. Epub 2017 Nov 6.
In the present study, the relative distribution of endophytic rhizobia in field-collected root nodules of the promiscuous host mung bean was investigated by sequencing of 16S ribosomal RNA (rRNA) and nifH genes, amplified directly from the nodule DNA. Co-dominance of the genera Bradyrhizobium and Ensifer was indicated by 32.05 and 35.84% of the total retrieved 16S rRNA sequences, respectively, and the sequences of genera Mesorhizobium and Rhizobium comprised only 0.06 and 2.06% of the recovered sequences, respectively. Sequences amplified from rhizosphere soil DNA indicated that only a minor fraction originated from Bradyrhizobium and Ensifer strains, comprising about 0.46 and 0.67% of the total retrieved sequences, respectively. 16S rRNA gene sequencing has also identified the presence of several non-rhizobial endophytes from phyla Proteobacteria, Actinobacteria, Bacteroides, and Firmicutes. The nifH sequences obtained from nodules also confirmed the co-dominance of Bradyrhizobium (39.21%) and Ensifer (59.23%) strains. The nifH sequences of the genus Rhizobium were absent, and those of genus Mesorhizobium comprised only a minor fraction of the sequences recovered from the nodules and rhizosphere soil samples. Two bacterial isolates, identified by 16S rRNA gene sequence analysis as Bradyrhizobium strain Vr51 and Ensifer strain Vr38, successfully nodulated the original host (mung bean) plants. Co-dominance of Bradyrhizobium and Ensifer strains in the nodules of mung bean indicates the potential role of the host plant in selecting specific endophytic rhizobial populations. Furthermore, successful nodulation of mung bean by the isolates showed that strains of both the genera Bradyrhizobium and Ensifer can be used for production of inoculum.
在本研究中,通过直接从根瘤 DNA 扩增的 16S 核糖体 RNA(rRNA)和 nifH 基因测序,研究了田间收集的杂宿主绿豆根瘤内生菌的相对分布。32.05%和 35.84%的总回收 16S rRNA 序列分别指示布拉氏菌属和固氮螺菌属的共优势,而 Mesorhizobium 和 Rhizobium 属的序列仅分别占回收序列的 0.06%和 2.06%。从根际土壤 DNA 扩增的序列表明,只有一小部分源自布拉氏菌属和固氮螺菌属菌株,分别占总回收序列的约 0.46%和 0.67%。16S rRNA 基因测序还鉴定了来自变形菌门、放线菌门、拟杆菌门和厚壁菌门的几种非根瘤内生菌。从根瘤中获得的 nifH 序列也证实了布拉氏菌属(39.21%)和固氮螺菌属(59.23%)菌株的共优势。根瘤中 Rhizobium 属的 nifH 序列不存在,而 Mesorhizobium 属的 nifH 序列仅占从根瘤和根际土壤样品中回收序列的一小部分。通过 16S rRNA 基因序列分析鉴定的两个细菌分离株,Bradyrhizobium 菌株 Vr51 和 Ensifer 菌株 Vr38,成功地接种了原始宿主(绿豆)植物。绿豆根瘤中布拉氏菌属和固氮螺菌属菌株的共优势表明,宿主植物在选择特定的内生根瘤菌种群方面发挥了作用。此外,分离株对绿豆的成功接种表明,布拉氏菌属和固氮螺菌属的菌株都可用于生产接种剂。