桑树“无菌”芽离体培养菌落细菌的分离与鉴定

Mamoru Sato, Wei Wei, H. Kawakita, H. Yamanouchi
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引用次数: 0

摘要

尽管在MS培养基上种植前对培养源(芽)进行了灭菌,但在离体培养的桑苗底部定期观察到细菌样菌落。从这些菌落中分离出5株细菌并进行了鉴定。所有菌株均在LB培养基上形成黄色菌落,命名为Y1-Y5,但在MS培养基上均未生长。根据病原菌的基本细菌学特性、API-20NE鉴定试剂盒和16S r-RNA基因序列分析,鉴定该菌株为油菜黄单胞菌。将菌株Y1 (RSF1010质粒作为标记物引入到菌株Y1中)接种到离体培养的桑树芽中,16天后,桑树芽底部出现了类似类型的菌落。菌株Yl (RSF1010)在接种植株45天后也能从大部分叶片中分离到。这些发现表明,该菌株在植物组织中存活了很长时间。因此,证实了桑树芽中渗出的菌落起源于逃脱表面灭菌的芽内的X, campestris。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Isolation and characterization of bacteria from colonies exuded from “germ-free” mulberry shoots cultured in vitro
Bacteria-like colonies have been periodically observed to exude from the bottom of mulberry shoots grown in vitro, although the culture origins (buds) were sterilized before planting on MS medium. From these colonies, five bacterial strains were isolated and characterized. All strains, designated as Y1-Y5, formed yellow colonies on LB medium, but none of them grew on MS medium. The present strains were identified as Xanthomonas campestris on the basis of principle bacteriological properties and the analyses of API-20NE identification kit and 16S r-RNA gene sequence. When strain Y1 (RSF1010) (RSF1010 plasmid was introduced as a marker into strain Y1) was inoculated into the mulberry shoots grown in vitro, similar types of colonies appeared at the bottom of the shoots after 16 days. Strain Yl (RSF1010) was also isolated from most of the leaves of the inoculated plants after as long as 45 days. These findings suggest that the strain survives within plant tissues for a long time. Thus, it was verified that colonies exuded from mulberry shoots originated from X, campestris within buds that escaped surface-sterilization.
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