南方农业学报2013，Vol.44Issue(1)：1-5,5.DOI:10.3969/j:issn.2095-1191.2013.1.1
甘蓝黑腐病菌XC_3374基因功能的初步探究
Preliminary study on the function of Xanthomone campestris pv.campestris XC_3374 gene
摘要
Abstract
[Objective]The functions of XC_3374 gene in Xanthomonas campestris pv. campestris Xcc8004 strain were preliminarily studied in order to lay the foundation for future studies of L-asparaginase function in Xcc8004. [Method]From the flanking sequence of XC_3374 gene obtained from the KEGG database, through conventional PCR methods, the genes were cloned, and at the same time, using the Dutch act plasmid pK18mobSacB through the homologous double exchange method, the mutant DM3374 was constructed and its phenotype was detected. [Result]By the D3374L-F/D3374R-R primer amplification, wild strains could amplify 1789 bp DNA fragments, while the mutant strains could amplify 1367 bp fragments. Using the amplified internal primers 3374F/3374R, wild strains could amplify 332 bp fragments, while the mutant strains could amplify fragments of any length, which showed that the target gene has been deleted, so the DM3374 deletion mutant was constructed successfully. The results showed that through the plate detection method, mutated XC_3374 gene did not affect the exopolysaccharides and extracellular enzyme synthesis or secretion. On top of the MMX basic medium culture, the mutant strains could grow normally and the colony size and form were basically the same as the wild strains, which indicated that the deletion mutant was not auxotrophic. The moving plate test results showed that the exercise capacity of XC_3374 deletion mutant was better than that of the wild strains, but the difference was not significant. Other test results showed that by using the clipping method for detecting deletion mutant's pathogenicity, the average length of the scabs in wild Xcc8004 and deletion mutant DM3374 were respectively 13.000 and 11.983 mm, and there was no significant difference between the two strains. [Conclusion]In the synthesis of extracellular polysaccharide and several extracellular enzymes, the secretion and pathogenic phenotype of XC_3374 mutant strains were consistent with those of the wild strains.关键词
甘蓝黑腐病菌/Xcc8004/XC_3374基因/突变体/表型分析/L-天冬酰胺酶Key words
Xanthomonas campestris pv. campestris/ Xcc8004/ XC_3374 gene/ mutant/ phenotypic analysis/ L-asparaginase分类
农业科技引用本文复制引用
梁伟,白先放,徐玉婷,李磊,安世琦,陆光涛..甘蓝黑腐病菌XC_3374基因功能的初步探究[J].南方农业学报,2013,44(1):1-5,5.基金项目
国家自然科学基金项目(30771177) （30771177）
广西亚热带生物资源保护与利用重点实验室项目(SB0705) （SB0705）
