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解淀粉芽孢杆菌B9601-Y2抗性基因标记及其在作物根部的定殖能力
引用本文:王志远,吴兴兴,吴毅歆,毛自朝,何月秋.解淀粉芽孢杆菌B9601-Y2抗性基因标记及其在作物根部的定殖能力[J].华中农业大学学报,2012,31(3):313-319.
作者姓名:王志远  吴兴兴  吴毅歆  毛自朝  何月秋
作者单位:1. 农业生物多样性应用技术国家工程研究中心,昆明,650201
2. 云南农业大学农学与生物技术学院,昆明,650201
3. 农业生物多样性应用技术国家工程研究中心,昆明650201;云南农业大学农学与生物技术学院,昆明650201
基金项目:国家自然科学基金项目(30660007)和科技部国际科技合作项目(2009DFA32360)
摘    要:以质粒pMarA转入解淀粉芽孢杆菌植生亚种B9601-Y2菌株,获得了卡那霉素抗性标记遗传稳定的菌株Y2-pMarA。采用浇灌法、拌种法和浸根法接种大白菜,并用浇灌法接种烟草,观察菌株在大白菜和烟草根部的定殖与消长动态。结果表明:通过10μg/mL卡那霉素的LB平板和PCR方法,证明标记菌株Y2-pMarA均能在植株根际、根表和根内定殖。在播种时浇灌大白菜37d后,在自然土处理中根际土、根表土和根内菌量分别为浇灌7d后菌量的94.31%、95.00%和84.75%,在灭菌土中则分别为75.26%、84.92%和177.74%;在拌种处理大白菜时,自然土处理中根际土、根表土和根内菌量分别为拌种7d后菌量的94.44%、95.27%和61.06%,在灭菌土处理中则分别为75.64%、90.91%和69.06%;在浸根10min后移栽,标记菌株能在大白菜根际土壤中扩散和进入根内;至移栽后33d,自然土处理中根际土、根表土和根内菌株定殖密度分别为6.28×105、9.54×105和4.69×103 cfu/g;灭菌土处理中则分别为6.97×105、1.12×106和1.02×104 cfu/g。

关 键 词:穿梭质粒  标记菌株  定殖  生防菌株
收稿时间:4/8/2011 12:00:00 AM

Resistance genes labeling and colonization ability of a biocontrol agent B9601-Y2 of Bacillus amyloliquefaciens in crop rhizospheres
WANG Zhi-yuan , WU Xing-xing , WU Yi-xin , MAO Zi-chao , HE Yue-qiu.Resistance genes labeling and colonization ability of a biocontrol agent B9601-Y2 of Bacillus amyloliquefaciens in crop rhizospheres[J].Journal of Huazhong Agricultural University,2012,31(3):313-319.
Authors:WANG Zhi-yuan  WU Xing-xing  WU Yi-xin  MAO Zi-chao  HE Yue-qiu
Institution:1,2 1.National Engineering Center of Agricultural Biodiversity,Kunming 650201,China; 2.College of Agronomy and Biotechnology,Yunnan Agricultural University, Kunming 650201,China
Abstract:The kanamycin-resistant labeling strain Y2-pMarA,which has a good stability of genetics,was gained by electrotransformation of shuttle plasmid pMarA into Bacillus amyloliquefaciens subsp.plantarum B9601-Y2.Chinese cabbage was inoculated by drenching,coating seeds,soaking root and tobacco by drenching.The results show as follows: LB agar plate containing 10 μg/mL of kanamycin and PCR proved that Y2-pMarA could colonize in rhizosphere soil,root surface and inner root.After seeding for 37 d,the Chinese cabbage,under the natural soil condition,colonization numbers of rhizosphere soil,root surface,and inner root of natural soil were 94.31%,95.00% and 84.75% of seeding for 7 d respectively;the corresponding ratios in sterilized soil were 75.26%,84.92% and 177.74% respectively.In the Chinese cabbage with coating seeds,the ratios of colonization numbers of rhizosphere soil,root surface,and inner root of natural soil were 94.44%,95.27% and 61.06% of seeding for 7 d respectively,while they were 75.6%,90.9% and 69.1% in sterilized soil respectively.By transplanting Chinese cabbage seedlings soaked for 10 min,Y2-pMarA strain could expand into the soil and enter the roots of the plant.After transplanting for 33 d,the colonization density in the rhizosphere soil,root surface and inner root in the natural soil were 6.28×105,9.54×105 and 4.69×103 cfu/g respectively;while the corresponding density were 6.97×105,1.12×106 and 1.02×104 cfu/g in sterilized soil respectively.
Keywords:shuttle plasmid  labeling strain  colonization  biocontrol agent
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