大麻(Cannabis sativa L.)内生细菌的分离与鉴定

Isolation and Identification of Endophytic Bacteria in Cannabis sativa L.

为分离与鉴定大麻(Cannabis sativa L.)雌株及雄株3个部位的叶片(从植株茎基向茎尖方向第二轮叶,第五轮叶,雄花周围叶)和花的内生细菌,本研究采用微生物培养和分子生物学鉴定方法,探讨其种类及其分布特征。结果显示,从大麻雄株中分离到6属35株内生细菌,大麻雌株中分离到11属39株内生细菌。内生细菌种属在大麻雌雄植株不同部位的叶和花中的分布存在显著差异,大麻雌株与雄株的内生细菌种类只共有葡萄球菌属(Staphylococcus sp.)、芽孢杆菌属(Bacillus sp.)和马赛菌属(Massilia sp.),其余均不共有。大麻雄株3个部位叶片中共有阿尔莱特葡萄球菌和溶血葡萄球菌而雄花中不存在;在雄花中分离到地衣芽孢杆菌、木糖葡萄球菌和贝莱斯芽孢杆菌而叶中不存在。大麻雌株3个部位叶片共有贝莱斯芽孢杆菌和特基拉芽孢杆菌而雌花中不存在;在雌花中分离到粪短波单胞菌、谷粒申氏菌、农杆菌、香料葡萄球菌、缺陷短波单胞菌、地衣芽胞杆菌和弗留明拜叶林克氏菌而叶中不存在。结果表明大麻内生细菌在不同植株、叶和花中的多样性以及不同内生细菌的分布差异。本研究结果为进一步研究大麻内生细菌的多样性提供科学依据,同时为未来大麻内生菌功能性促生菌种的筛选提供基础。

In order to isolate and identify the endophytic bacteria of the female and male plants of Cannabis sativa L.in three parts of the leaves(the second round leaf,the fifth round leaf,and the leaves around the male flower)and the flower.In this study,microbial culture and molecular biology identification methods were used to explore their types and distribution characteristics.The results showed that 35 endophytic bacteria of 6 genera were isolated from male cannabis plants,and 39 endophytic bacteria of 11 genera were isolated from female Cannabis sativa L..There are significant differences in the distribution of endophytic bacterial species in the leaves and flowers of different parts of the male and female Cannabis sativa L..The endophytic bacterial species of the female and male Cannabis sativa L.only share Staphylococcus sp.and Bacllus sp.and Massilia sp.,the rest are not shared.Staphylococcus arlettae and Staphylococcus haemolyticus are shared in the three leaves of the male Cannabis sativa L.,but do not exist in the male flower.In male flowers,Bacillus licheniformis,Staphylococcus xylosus and Bacillus velezensis were isolated but not in leaves.The three leaves of the female Cannabis sativa L.share Bacillus velezensis and Bacillus tequilensis and do not exist in the female flower.Brevundimonas faecalis,Shinella granuli,Agrobacterium tumefaciens,Staphylococcus condiment,Brevundimonas diminuta,Bacillus licheniformis,and Beijerinckia fluminensis were isolated in female flowers and did not exist in leaves.The results showed that the diversity of Cannabis sativa L.endophytic bacteria in different plants,leaves and flowers,and the differences in the distribution of different endophytic bacteria.The results of this study provide a scientific basis for further research on the diversity of Cannabis sativa L.endophytic bacteria and lay a foundation for the future screening of functional growth-promoting bacteria of Cannabis sativa L.endophytes.