Kamoun S, Kado C I
Department of Plant Pathology, University of California, Davis 95616.
J Bacteriol. 1990 Sep;172(9):5165-72. doi: 10.1128/jb.172.9.5165-5172.1990.
Using Tn4431, a transposon that allows transcriptional fusions to a promoterless luciferase (lux) operon, we have isolated a nonpathogenic mutant of Xanthomonas campestris pv. campestris, i.e., JS111, that does not incite any of the black rot symptoms on all tested cruciferous host plants (J. J. Shaw, L. G. Settles, and C. I. Kado, Mol. Plant Microbe Interact. 1:39-45, 1988). In the study reported here, we determined that in contrast to the wild-type strain, JS111 is unable to induce a hypersensitive necrotic response on nonhost plants such as datura, tomato, and cucumber, suggesting that JS111 is a nonpathogenic, nonhypersensitive Hrp mutant. JS111 displayed culture growth rates, exopolysaccharide production, and protease, pectate lysase, cellulase, amylase, and phosphatase activities comparable to those of the wild-type strain. However, the growth of JS111 in host leaves was markedly attenuated. Coinoculation of JS111 with the wild-type strain in cauliflower or radish leaves rescued the growth deficiency of the mutant to normal levels. The locus mutated in JS111 was cloned and named hrpXc, and transcriptional and genetic complementation analyses of the hrpXc locus were conducted. The regulation of hrpXc expression was also investigated in vitro and in planta, using fusions to a lux or chloramphenicol acetyltransferase reporter gene. The hrpXc gene was found to be strongly induced in radish leaves. This is the first report and analysis of a hrp locus from a Xanthomonas species.
利用转座子Tn4431(一种可使无启动子的荧光素酶(lux)操纵子发生转录融合的转座子),我们分离出了野油菜黄单胞菌野油菜致病变种的一个非致病突变体,即JS111,它在所有测试的十字花科寄主植物上均不引发任何黑腐病症状(J. J. 肖、L. G. 塞茨和C. I. 卡多,《分子植物与微生物相互作用》1:39 - 45,1988年)。在本文报道的研究中,我们确定,与野生型菌株相比,JS111无法在非寄主植物如曼陀罗、番茄和黄瓜上诱导超敏坏死反应,这表明JS111是一个非致病、非超敏的Hrp突变体。JS111的培养生长速率、胞外多糖产生以及蛋白酶、果胶酸裂解酶、纤维素酶、淀粉酶和磷酸酶活性与野生型菌株相当。然而,JS111在寄主叶片中的生长明显减弱。将JS111与野生型菌株共同接种在花椰菜或萝卜叶片中,可使突变体的生长缺陷恢复到正常水平。克隆了JS111中发生突变的位点并命名为hrpXc,并对hrpXc位点进行了转录和遗传互补分析。还利用与lux或氯霉素乙酰转移酶报告基因的融合,在体外和植物体内研究了hrpXc表达的调控。发现hrpXc基因在萝卜叶片中被强烈诱导。这是关于黄单胞菌属hrp位点的首次报道和分析。