Department of Genetics, Harvard Medical School, Boston, MA, USA.
Proc Natl Acad Sci U S A. 1985 Sep;82(17):5841-5. doi: 10.1073/pnas.82.17.5841.
Parasponia, a woody member of the elm family, is the only nonlegume genus whose members are known to form an effective nitrogen-fixing symbiosis with a Rhizobium species. The bacterial strain RP501 is a slow-growing strain of Rhizobium isolated from Parasponia nodules. Strain RP501 also nodulates the legumes siratro (Macroptilium atropurpureum) and cowpea (Vigna unguiculata). Using a cosmid clone bank of RP501 DNA, we isolated a 13.4-kilobase (kb) EcoRI fragment that complemented insertion and point mutations in three contiguous nodulation genes (nodABC) of Rhizobium meliloti, the endosymbiont of alfalfa (Medicago sativa). The complemented R. meliloti nod mutants induced effective nitrogen-fixing nodules on alfalfa seedlings but not on siratro, cowpeas, or Parasponia. The cloned RP501 nodulation locus hybridized to DNA fragments carrying the R. meliloti nodABC genes. A 3-kb cluster of Tn5 insertion mutations on the RP501 13.4-kb EcoRI fragment prevented complementation of R. meliloti nodABC mutations.
Parasponia 是榆科的木本植物,是唯一已知与 Rhizobium 物种形成有效固氮共生关系的非豆科属。细菌菌株 RP501 是从 Parasponia 根瘤中分离出来的一种生长缓慢的 Rhizobium 菌株。菌株 RP501 还能结瘤豆科植物豇豆(Macroptilium atropurpureum)和豇豆(Vigna unguiculata)。我们使用 RP501 DNA 的 cosmid 克隆库,分离出一个 13.4 千碱基(kb)的 EcoRI 片段,该片段可互补 Rhizobium meliloti 中三个连续的结瘤基因(nodABC)的插入和点突变,Rhizobium meliloti 是紫花苜蓿(Medicago sativa)的内共生体。互补的 R. meliloti nod 突变体在紫花苜蓿幼苗上诱导有效固氮结瘤,但在豇豆、豇豆或 Parasponia 上则没有。克隆的 RP501 结瘤基因座与携带 R. meliloti nodABC 基因的 DNA 片段杂交。在 RP501 13.4kb EcoRI 片段上的 Tn5 插入突变簇阻止了 R. meliloti nodABC 突变的互补。