Department of Soil Science and Department of Microbiology, University of Minnesota, St. Paul, Minnesota 55108; Nitrogen Fixation and Soybean Genetics Laboratory, Agriculture Research Service, U.S. Department of Agriculture, Beltsville, Maryland 20705 ; and The NifTAL Project, University of Hawaii, Paia, Hawaii 96779.
Appl Environ Microbiol. 1990 Jun;56(6):1768-74. doi: 10.1128/aem.56.6.1768-1774.1990.
Several soybean plant introduction (PI) genotypes have recently been described which restrict nodulation of Bradyrhizobium japonicum serocluster 123 in an apparently serogroup-specific manner. While PI 371607 restricts nodulation of strains in serogroup 123 and some in serogroup 127, those in serogroup 129 are not restricted. When DNA regions within and around the B. japonicum I-110 common nodulation genes were used as probes to genomic DNA from the serogroup strains USDA 123, USDA 127, and USDA 129, several of the probes differentially hybridized to the nodulation-restricted and -unrestricted strains. One of the gene regions, cloned in plasmid pMJS12, was subsequently shown to hybridize to 4.6-kilobase EcoRI fragments from DNAs from nodulation-restricted strains and to larger fragments in nodulation-unrestricted strains. To determine if the different hybridization patterns could be used to predict nodulation restriction, we hybridized pMJS12 to EcoRI-digested genomic DNAs from uncharacterized serocluster 123 field isolates. Of the 36 strains examined, 15 were found to have single, major, 4.6-kilobase hybridizing EcoRI fragments. When tested for nodulation, 80% (12 of 15) of the strains were correctly predicted to be restricted for nodulation of the PI genotypes. In addition, hybridization patterns obtained with pMJS12 and nodulation phenotypes on PI 371607 indicated that there are at least three types of serogroup 127 strains. Our results suggest that the pMJS12 gene probe may be useful in selecting compatible host-strain combinations and in determining the suitability of field sites for the placement of soybean genotypes containing restrictive nodulation alleles.
几种大豆引种(PI)基因型最近被描述为以明显的血清群特异性方式限制 Bradyrhizobium japonicum 血清群 123 的结瘤。虽然 PI 371607 限制了血清群 123 和一些血清群 127 中的菌株的结瘤,但血清群 129 中的菌株不受限制。当将 B. japonicum I-110 共同结瘤基因的内部和周围的 DNA 区域用作探针,对血清群菌株 USDA 123、USDA 127 和 USDA 129 的基因组 DNA 进行探测时,一些探针与结瘤受限和不受限的菌株杂交。其中一个基因区域,克隆在质粒 pMJS12 中,随后显示与结瘤受限菌株的 4.6 千碱基 EcoRI 片段杂交,并与结瘤不受限菌株的较大片段杂交。为了确定不同的杂交模式是否可用于预测结瘤限制,我们将 pMJS12 与未表征的血清群 123 田间分离株的 EcoRI 消化基因组 DNA 杂交。在所检查的 36 株中,有 15 株具有单一的、主要的 4.6 千碱基杂交 EcoRI 片段。当对结瘤性进行测试时,80%(15 株中的 12 株)的菌株被正确预测为对 PI 基因型的结瘤受到限制。此外,用 pMJS12 进行杂交和在 PI 371607 上的结瘤表型表明,至少有三种类型的血清群 127 菌株。我们的结果表明,pMJS12 基因探针可能有助于选择相容的宿主-菌株组合,并确定含有限制结瘤等位基因的大豆基因型的放置适用于现场站点。