春兰根内生细菌分离培养方法的初步研究  

Primary research on the isolation method of root endophytic bacteria of Cymbidium goeringii

作　　者：孙磊[1] 毕晓宝[1,2] 李潞滨[3] 刘琳[1] 韩继刚[1] 杨凯[4] 

机构地区：[1]河北大学生命科学学院,河北保定071002 [2]中国医学科学院北京协和医学院药用植物研究所,北京100094 [3]中国林业科学研究院林业研究所,国家林业局林木培育重点实验室,北京100091 [4]北京农学院,农业应用新技术北京市重点实验室,北京102206

出　　处：《河北农业大学学报》2009年第1期42-46,共5页Journal of Hebei Agricultural University

基　　金：“948”项目(2009-4-C08);国家“十一五”科技支撑计划课题之林业科技支撑专题(2006BAD01A18);河北大学引进人才专项基金(2006-088)

摘　　要：探讨了春兰根表面灭菌的最佳条件,以及改良察氏琼脂、淀粉铵营养琼脂、YG、R2A、LB、金氏、TSA和根系分泌物8种培养基对春兰根内生细菌的分离效果。结果表明:春兰根的最佳表面灭菌条件为:75%乙醇浸泡3 min,3%次氯酸钠溶液处理2 min,75%乙醇浸泡30 s,最后用无菌水冲洗。不同培养基对内生细菌的分离存在一定差异,R2A培养基和根系分泌物培养基分离出的内生细菌数量较多,而YG培养基和R2A培养基分离出的内生细菌种类总数最多。此8种培养基对春兰根内生细菌的优势菌群的分离效果是相同的。研究结果可为选择合适的表面灭菌条件及培养基分离兰花内生细菌提供参考。This paper studies the optimal surface sterilization condition of Cymbidium goeringii Rchb. f. roots and their effect on endophytic bacteria isolated by eight media. The 8 media include Modified czapek agar, Starch-ammonium nutrient agar, YG medium, R2A medium, Luria- Bertani medium （LB）, King＇s medium B （KMB）, Tryptic Soy Agar （TSA） and Root exudate medium. The results indicate that conditions for best surface sterilization were： roots washed with 75 % ethanol for 3 min, immersed in fresh sodium hypochlorite solution （3% available Cl^- ） for 2 min, rinsed with 75 % ethanol for 30 s, and finally washed with sterile distilled water. There were certain differences among the endophytic bacteria isolated by using different media. CFU on R2A medium and root exudate medium outnumbered that on the other media, and the most kinds of endophytic bacteria were found on YG and R2A media. 16S rDNA sequences of representative strains from different media showed high similarity with Rhizobium spp. , suggesting that the eight media had the same effect on the isolated dominant endophytic group. The results may contribute to isolation of orchid endophytic bacteria by using proper medium.

关 键 词：春兰 内生细菌 表面灭菌 培养基 分离 

分 类 号：S858.28[农业科学—临床兽医学] TS251.1[农业科学—兽医学]