Department of Biotechnology, Tshwane University of Technology, Pretoria, South Africa.
BMC Microbiol. 2010 Mar 23;10:89. doi: 10.1186/1471-2180-10-89.
Cowpea is the most important food grain legume in Sub-Saharan Africa. However, no study has so far assessed rhizobial biodiversity and/or nodule functioning in relation to strain IGS types at the continent level. In this study, 9 cowpea genotypes were planted in field experiments in Botswana, South Africa and Ghana with the aim of i) trapping indigenous cowpea root-nodule bacteria (cowpea "rhizobia") in the 3 countries for isolation, molecular characterisation using PCR-RFLP analysis, and sequencing of the 16S - 23S rDNA IGS gene, ii) quantifying N-fixed in the cowpea genotypes using the 15N natural abundance technique, and iii) relating the levels of nodule functioning (i.e. N-fixed) to the IGS types found inside nodules.
Field measurements of N2 fixation revealed significant differences in plant growth, delta15N values, %Ndfa and amounts of N-fixed between and among the 9 cowpea genotypes in Ghana and South Africa. Following DNA analysis of 270 nodules from the 9 genotypes, 18 strain IGS types were found. Relating nodule function to the 18 IGS types revealed significant differences in IGS type N2-fixing efficiencies. Sequencing the 16S - 23S rDNA gene also revealed 4 clusters, with cluster 2 forming a distinct group that may be a new Bradyrhizobium species. Taken together, our data indicated greater biodiversity of cowpea bradyrhizobia in South Africa relative to Botswana and Ghana.
We have shown that cowpea is strongly dependant on N2 fixation for its N nutrition in both South Africa and Ghana. Strain IGS type symbiotic efficiency was assessed for the first time in this study, and a positive correlation was discernible where there was sole nodule occupancy. The differences in IGS type diversity and symbiotic efficiency probably accounts for the genotype x environment interaction that makes it difficult to select superior genotypes for use across Africa. The root-nodule bacteria nodulating cowpea in this study all belonged to the genus Bradyrhizobium. Some strains from Southern Africa were phylogenetically very distinct, suggesting a new Bradyrhizobium species.
豇豆是撒哈拉以南非洲最重要的粮食豆科作物。然而,迄今为止,还没有研究评估过与菌株 IGS 类型相关的根瘤菌生物多样性和/或结瘤功能在整个非洲大陆的情况。在这项研究中,在博茨瓦纳、南非和加纳的田间试验中种植了 9 个豇豆品种,目的是:i)在 3 个国家中捕获本地豇豆根瘤菌(豇豆“根瘤菌”)进行分离,使用 PCR-RFLP 分析进行分子特征分析,并对 16S-23S rDNA IGS 基因进行测序,ii)使用 15N 自然丰度技术定量测定豇豆品种中的固氮量,iii)将结瘤功能(即固氮)与在结瘤中发现的 IGS 类型相关联。
对 N2 固定的田间测量表明,在加纳和南非的 9 个豇豆品种之间和品种内的植物生长、delta15N 值、%Ndfa 和固氮量存在显著差异。对 9 个品种的 270 个结瘤进行 DNA 分析后,发现了 18 种菌株 IGS 类型。将结瘤功能与 18 种 IGS 类型相关联表明,IGS 类型的固氮效率存在显著差异。对 16S-23S rDNA 基因进行测序也揭示了 4 个聚类,其中聚类 2 形成了一个可能是新的慢生根瘤菌属的独特群体。总的来说,我们的数据表明,与博茨瓦纳和加纳相比,南非豇豆根瘤菌的生物多样性更大。
我们已经表明,在南非和加纳,豇豆对其氮营养的强烈依赖于 N2 固定。在这项研究中,首次评估了菌株 IGS 类型共生效率,并发现了一个正相关,其中存在单一的结瘤占据。IGS 类型多样性和共生效率的差异可能解释了基因型与环境相互作用的原因,这使得难以在整个非洲选择优良的基因型。本研究中结瘤豇豆的根瘤菌均属于慢生根瘤菌属。来自南非的一些菌株在系统发育上非常独特,表明可能存在一个新的慢生根瘤菌属。