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Transfer of the Pea Symbiotic Plasmid pJB5JI in Nonsterile Soil

机译:豌豆共生质粒pJB5JI在非无菌土壤中的转移

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Transfer of the pea (Pisum sativum L.) symbiotic plasmid pJB5JI between strains of rhizobia was examined in sterile and nonsterile silt loam soil. Sinorhizobium fredii USDA 201 and HH003 were used as plasmid donors, and symbiotic plasmid-cured Rhizobium leguminosarum 6015 was used as the recipient. The plasmid was carried but not expressed in S. fredii strains, whereas transfer of the plasmid to R. leguminosarum 6015 rendered the recipient capable of nodulating pea plants. Confirmation of plasmid transfer was obtained by acquisition of plasmid-encoded antibiotic resistance genes, nodulation of pea plants, and plasmid profiles. Plasmid transfer in nonsterile soil occurred at frequencies of up to 10?4 per recipient and appeared to be highest at soil temperatures and soil moisture levels optimal for rhizobial growth. Conjugation frequencies were usually higher in sterile soil than in nonsterile soil. In nonsterile soil, transconjugants were recovered only with strain USDA 201 as the plasmid donor. Increasing the inoculum levels of donor and recipient strains up to 109 cells g of soil?1 increased the number of transconjugants; peak plasmid transfer frequencies, however, were found at the lower inoculum level of 107 cells g of soil?1. Plasmid transfer frequencies were raised in the presence of the pea rhizosphere or by additions of plant material. Transconjugants formed by the USDA 201(pJB5JI) × 6015 mating in soil formed effective nodules on peas.
机译:在无菌和非无菌淤泥壤土中检查了豌豆(Pisum sativum L.)共生质粒pJB5JI在根瘤菌菌株之间的转移。 fredii中华根瘤菌USDA 201和HH003被用作质粒供体,共生质粒固化的豆科根瘤菌6015被用作受体。携带质粒但未在弗氏链球菌菌株中表达,而将质粒转移至豆科菌根瘤菌6015使受体能够结瘤豌豆植物。通过获取质粒编码的抗生素抗性基因,豌豆植株的结节和质粒图谱来获得质粒转移的确认。在非无菌土壤中,质粒转移的发生频率高达每位受体10?4,在土壤温度和最适合根瘤菌生长的土壤水分水平下似乎最高。无菌土壤中的共轭频率通常高于非无菌土壤。在非无菌土壤中,仅用USDA 201菌株作为质粒供体回收了转结合剂。将供体和受体菌株的接种量提高到109细胞/克土壤?1,可增加转结合子的数量。然而,在较低的接种量水平下,在107细胞/克土壤中发现了最大的质粒转移频率。在豌豆根际存在下或通过添加植物材料来提高质粒转移频率。 USDA 201(pJB5JI)×6015在土壤中交配形成的共轭结合物在豌豆上形成有效结节。

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