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
余亿 1普凤雅 1王兵兵 1黄军 2杨艺婷 1唐琴 1杨志清3
作者信息
- 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 农学与生物技术学院,云南 昆明 650201;云南农业大学 特色小宗作物研究中心,云南 昆明 650201;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
- 折叠
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.
Abstract
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Key words
Coix lacryma-jobi L./Bacillus velezensis/Colonization ability/Plasmid引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据