Minsavage G V, Mudgett M B, Stall R E, Jones J B
Plant Pathology Department, University of Florida, PO Box 110680, Gainesville, FL 32611, USA.
Mol Plant Microbe Interact. 2004 Feb;17(2):152-61. doi: 10.1094/MPMI.2004.17.2.152.
Tn5 insertion mutants of Xanthomonas campestris pv. vesicatoria were inoculated into tomato and screened for reduced virulence. One mutant exhibited reduced aggressiveness and attenuated growth in planta. Southern blot analyses indicated that the mutant carried a single Tn5 insertion not associated with previously cloned pathogenicity-related genes of X. campestris pv. vesicatoria. The wild-type phenotype of this mutant was restored by one recombinant plasmid (pOPG361) selected from a genomic library of X. campestris pv. vesicatoria 91-118. Tn3-gus insertion mutagenesis and sequence analyses of a subclone of pOPG361 identified a 1,929-bp open reading frame (ORF) essential for complementation of the mutants. The predicted protein encoded by this ORF was highly homologous to the previously reported pathogenicity-related HrpM protein of Pseudomonas syringae pv. syringae and OpgH of Erwinia chrysanthemi. Based on homology, the new locus was designated opgHXcv. Manipulation of the osmotic potential in the intercellular spaces of tomato leaves by addition of mannitol at low concentrations (25 to 50 mM) compensates for the opgHXcv mutation.
将野油菜黄单胞菌致病变种的Tn5插入突变体接种到番茄中,并筛选毒力降低的突变体。一个突变体在植物中的侵袭力降低且生长减弱。Southern杂交分析表明,该突变体携带一个单一的Tn5插入片段,与野油菜黄单胞菌致病变种先前克隆的致病性相关基因无关。从野油菜黄单胞菌致病变种91-118的基因组文库中筛选出的一个重组质粒(pOPG361)恢复了该突变体的野生型表型。对pOPG361的一个亚克隆进行Tn3-gus插入诱变和序列分析,鉴定出一个对突变体互补至关重要的1929 bp开放阅读框(ORF)。该ORF编码的预测蛋白与先前报道的丁香假单胞菌丁香致病变种的致病性相关HrpM蛋白和菊欧文氏菌的OpgH高度同源。基于同源性,将这个新基因座命名为opgHXcv。通过添加低浓度(25至50 mM)的甘露醇来调节番茄叶片细胞间隙中的渗透势,可以补偿opgHXcv突变。