Chen L S, Figueredo A, Pedrosa F O, Hungria M
Universidade Estadual de Londrina, PR, Brazil.
Appl Environ Microbiol. 2000 Nov;66(11):5099-103. doi: 10.1128/AEM.66.11.5099-5103.2000.
The soybean is an exotic plant introduced in Paraguay in this century; commercial cropping expanded after the 1970s. Inoculation is practiced in just 15 to 20% of the cropping areas, but root nodulation occurs in most sites where soybeans grow. Little is known about rhizobial diversity in South America, and no study has been performed in Paraguay until this time. Therefore, in this study, the molecular characterization of 78 rhizobial isolates from soybean root nodules, collected under field conditions in 16 sites located in the two main producing states, Alto Paraná and Itapúa, was undertaken. A high level of genetic diversity was detected by an ERIC-REP-PCR analysis, with the majority of the isolates representing unique strains. Most of the 58 isolates characterized by slow growth and alkaline reactions in a medium containing mannitol as a carbon source were clustered with strains representative of the Bradyrhizobium japonicum and Bradyrhizobium elkanii species, and the 16S ribosomal DNA (rDNA) sequences of 5 of those isolates confirmed the species identities. However, slow growers were highly polymorphic in relation to the reference strains, including five carried in commercial inoculants in neighboring countries, thus indicating that the Paraguayan isolates might represent native bradyrhizobia. Twenty isolates highly polymorphic in the ERIC-REP-PCR profiles were characterized by fast growth and acid reactions in vitro, and two of them showed high 16S rDNA identities with Rhizobium genomic species Q. However, two other fast growers showed high 16S rDNA identity with Agrobacterium spp., and both of these strains established efficient symbioses with soybean plants.
大豆是本世纪引入巴拉圭的外来植物;20世纪70年代后商业种植面积扩大。仅15%至20%的种植区域进行接种,但在大多数大豆种植地都有根瘤形成。关于南美洲根瘤菌多样性知之甚少,此前在巴拉圭尚未开展相关研究。因此,本研究对从位于两个主要生产州上巴拉那州和伊塔普阿州的16个地点的田间条件下收集的78株大豆根瘤根瘤菌分离株进行了分子特征分析。通过ERIC-REP-PCR分析检测到高度的遗传多样性,大多数分离株代表独特菌株。在以甘露醇作为碳源的培养基中生长缓慢且呈碱性反应的58株分离株中,大多数与日本慢生根瘤菌和埃氏慢生根瘤菌物种的代表性菌株聚类,其中5株分离株的16S核糖体DNA(rDNA)序列证实了物种身份。然而,与参考菌株相比,生长缓慢的菌株具有高度多态性,包括邻国商业接种剂中携带的5个菌株,因此表明巴拉圭分离株可能代表本地慢生根瘤菌。在ERIC-REP-PCR图谱中高度多态的20株分离株在体外生长迅速且呈酸性反应,其中2株与根瘤菌基因组物种Q的16S rDNA高度同源。然而,另外2株快速生长的菌株与土壤杆菌属的16S rDNA高度同源,且这两种菌株都与大豆植株建立了有效的共生关系。