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野油菜黄单胞菌野油菜致病变种8004菌株wxcA基因与EPS的产量有关
引用本文:陆光涛,唐纪良,危广宁,何勇强,陈保善.野油菜黄单胞菌野油菜致病变种8004菌株wxcA基因与EPS的产量有关[J].生物工程学报,2004,20(4):477-483.
作者姓名:陆光涛  唐纪良  危广宁  何勇强  陈保善
作者单位:广西大学分子遗传研究所,南宁,530005
基金项目:国家自然科学基金重点资助项目 (NO .3 0 13 0 0 10 )~~
摘    要:在以前的工作中,采用转座子Tn5 gusA5对野油菜黄单胞菌野油菜致病变种(Xcc)8004菌株进行诱变,获得一批胞外多糖(EPS)合成减少的突变体,对这些突变体的Tn5 gusA5的插入位点进行分析后,发现有两株突变体是wxcA基因不同插入位点的突变体。以前认为wxcA基因与脂多糖(LPS)的O-抗原合成有关而与EPS的合成无关。为明确wxc4基因的功能,对8004菌株的wxcA基因进行缺失,获得的△wxcA突变体的EPS产量与野生型菌株相比,减少了50%,并且一段PCR合成的包含wxcA基因的DNA片段能反式互补△wxcA突变体,恢复突变体的EPS产量。这证实了8004菌株的wxcA基因与EPS的合成产量有关。

关 键 词:野油菜黄单胞菌野油菜致病变种  wxcA基因  胞外多糖
文章编号:1000-3061(2004)04-0477-07
修稿时间:2003年12月13

The Gene wxcA of Xanthomonas campestris pv.campestris 8004 Strain Involved in EPS Yield
LU Guang Tao TANG Ji Liang,WEI Guang Ning HE Yong Qiang CHEN Bao Shan.The Gene wxcA of Xanthomonas campestris pv.campestris 8004 Strain Involved in EPS Yield[J].Chinese Journal of Biotechnology,2004,20(4):477-483.
Authors:LU Guang Tao TANG Ji Liang  WEI Guang Ning HE Yong Qiang CHEN Bao Shan
Institution:Institute of Molecular genetics, Guangxi University, Nanning 530005, China.
Abstract:Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.
Keywords:Xanthomonas campestris  pv    campestris  wxcA  gene  extracellular polysaccharide
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