Valinsky L, Manulis S, Nizan R, Ezra D, Barash I
Department of Plant Pathology, ARO, Volcani Center, Bet Dagan, Israel.
Mol Plant Microbe Interact. 1998 Aug;11(8):753-62. doi: 10.1094/MPMI.1998.11.8.753.
The host range of the gall-forming bacterium Erwinia herbicola pv. gypsophilae (Ehg) is restricted to the gypsophila plant whereas E. herbicola pv. betae (Ehb) incites galls on beet as well as gypsophila. The pathogenicity of Ehg and Ehb was previously shown to be dependent on a plasmid (pPATH). Transposon mutagenesis was used to generate mutants on the cosmid pLA150 of the pPATH from Ehg824-1. A cluster of nonpathogenic mutations flanked by two IS1327 elements was identified on a 3.2-kb NdeI DNA fragment. All mutants were restored to pathogenicity by complementation in trans with the wild-type Ehg DNA. DNA sequence analysis of the 3.2-kb NdeI fragment revealed a single open reading frame (ORF) of 2 kb as well as a potential ribosome binding site and a putative hrp box upstream to the ORF. The ORF had no significant homology to known genes. Southern analysis also revealed the presence of DNA sequences that hybridized to the ORF in the beet pathovar Ehb4188. This gene was isolated and sequenced. Marker exchange mutants generated in the ORF of Ehb eliminated the pathogenicity of Ehb on gypsophila but fully retained its pathogenicity on beet. Since the putative gene appeared to encode a host-specific virulence factor for gypsophila it was designated as hsvG.
形成虫瘿的细菌草生欧文氏菌嗜石竹变种(Ehg)的寄主范围仅限于石竹属植物,而草生欧文氏菌甜菜变种(Ehb)则能在甜菜和石竹上引发虫瘿。先前已证明Ehg和Ehb的致病性依赖于一种质粒(pPATH)。利用转座子诱变技术在来自Ehg824 - 1的pPATH粘粒pLA150上产生突变体。在一个3.2 kb的NdeI DNA片段上鉴定出了由两个IS1327元件侧翼的一组非致病性突变。所有突变体通过与野生型Ehg DNA进行反式互补而恢复致病性。对3.2 kb NdeI片段的DNA序列分析揭示了一个2 kb的单一开放阅读框(ORF),以及一个潜在的核糖体结合位点和一个位于该ORF上游的假定hrp框。该ORF与已知基因无明显同源性。Southern分析还揭示了在甜菜致病变种Ehb4188中存在与该ORF杂交的DNA序列。该基因被分离并测序。在Ehb的ORF中产生的标记交换突变体消除了Ehb对石竹的致病性,但完全保留了其对甜菜的致病性。由于该假定基因似乎编码一种石竹属植物特异性的毒力因子,因此将其命名为hsvG。