一株贝莱斯芽孢杆菌L21在薏苡植株体内的定殖研究  

Colonization of A Strain of Bacillus velezensis L21 in Coix lachryma-jobi

作　　者：余亿 普凤雅 王兵兵 黄军 杨艺婷 唐琴 杨志清[1,3,4] YU Yi;PU Feng-ya;WANG Bing-bing;HUANG Jun;YANG Yi-ting;TANG Qin;YANG Zhi-qing(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;Agricultural Technology Extension Center of Mengla County,Xishuangbanna Dai Autonomous Prefecture,Mengla 666300,China;Institute of Improvement and Utilization of Characteristic Resource Plants,Yunnan Agricultural University,Kunming 650201,China;National-Local Joint Engineering Research Center on Germplasms Utilization&Innovation of Chinese Medicinal Materials in Southwest China,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]云南省西双版纳傣族自治州勐腊县农业技术推广中心,云南勐腊666300 [3]云南农业大学特色小宗作物研究中心,云南昆明650201 [4]云南农业大学西南中药材种质创新与利用国家地方联合工程研究中心,云南昆明650201

出　　处：《江西农业学报》2024年第3期47-53,共7页Acta Agriculturae Jiangxi

基　　金：云南省科技计划项目(2014RE002)。

摘　　要：将含有抗生素的绿色荧光蛋白质粒标记在菌株L21(Bacillus velezensis)中,旨在深入研究内生细菌与薏苡植株的互作。采用自然转化法,将质粒pHT01-P43GFPmut3a转入薏苡内生菌L21中,获得了工程菌株L21-GFP,并测定了L21-GFP和L21的生长曲线、遗传稳定性、功能稳定性;用浇灌法将L21-GFP导入薏苡植株体内,并用荧光显微检测其定殖情况。结果表明:L21-GFP和L21的菌落形态、生长速度和拮抗活性之间无明显差异,均在培养8 h后从缓慢生长期进入对数生长期;pHT01-P43GFPmut3a能在L21中稳定遗传,转接40 h后的稳定性为63%;工程菌株L21-GFP能够稳定地定殖于薏苡植株体内,其中定殖于根部的菌体密度最大,达到106 cfu/g。由此可知,L21-GFP在薏苡体内定殖能力强且稳定,具有良好的应用前景。Bacillus velezensis(L21)was labeled with green fluorescent protein granules containing antibiotics,which provided theoretical basis for further study on interaction between endophytic bacteria and Coix lacrima-jobi.Firstly,plasmid pHT01-P43GFPmut3a was transferred into endophyte L21(Bacillus velezensis)by natural transformation method to obtain engineered strain L21-GFP.The growth curves,genetic stability and functional stability of L21-GFP and L21 were measured.L21-GFP was introduced into Coix lacrima-jobi by watering method,and the colonization was detected by fluorescence microscopy.The results showed that there was no significant difference in colony morphology,growth rate and antagonistic activity between L21-GFP and L21,both of which changed from slow growth stage to logarithmic growth stage after 8 h culture.pHT01-P43GFPmut3a can be stably inherited in L21,and the stability of pHT01-P43GFPmut3a is 63%after 40 h.The engineered strain L21-GFP was able to colonize stably in Coix lacrima-jobi,with the highest density of 106 cfu/g in the root.In conclusion,L21-GFP has strong and stable colonization ability in Coix lacrima-jobi,and has good application prospect.

关 键 词：薏苡 贝莱斯芽孢杆菌 定殖能力 质粒 

分 类 号：S543.9[农业科学—作物学]