椰纤果产生菌株分离鉴定和小型菌种库的建立  

Isolation,identification and library establishment of bacterial cellulose-producing strains in Nata de coco

作　　者：刘龙祥[1] 刘四新[1,2] 陈华美[1] 余敏华[1] 陈白羽[1] 毕继才[1] 李从发[1,2] 

机构地区：[1]海南大学食品学院,海南海口570228 [2]海南大学热带生物资源教育部重点实验室,海南海口570228

出　　处：《广东农业科学》2016年第3期142-147,共6页Guangdong Agricultural Sciences

基　　金：海南省自然科学基金(314048);海南省普通高校研究生创新科研项目(Hy2013-B02);海口市热带农产品深加工重点实验室项目(2013-45);海南大学研究生优秀学位论文培育计划项目

摘　　要：从海南省内外的椰纤果生产企业代表性的布点采样、分离以及交换等方式共获得145株细菌纤维素产生菌,对其中93株采用持家基因、16Sr DNA基因等序列分析结合形态特征进行鉴定,引入近缘的7株不同种的模式菌株作为参照。结果显示,这些菌株被鉴定为驹形氏杆菌属(Komagataeibacter),其中79株分别鉴定为3个已知种:K.nataicola、K.europaeus、K.hansenii,另14株为未知种。采用持家基因dna K、rpo B和gro EL对14株未知菌进行鉴定,发现它们两两序列相似度均大于种间差异的临界值99.0%、98.8%和99.2%,推测该14株菌可能为驹形氏杆菌属的同一个新种。实验室目前的细菌纤维素产生菌包含模式种在内至少有9个已知种、还有至少1个疑似新种。小型菌种库还在持续建设中,这为保护我国椰纤果生产的菌种资源、优良菌株的遗传选育、椰纤果发酵生产的技术升级以及细菌纤维素的相关理论研究等奠定了基础。145 bacterial cellulose-producing strains were obtained by isolation and collection from factories both at home and aboard of Hainan. 93 strains gained before 2015 were identified by phenotypic characteristics and molecular sequence features （ house-keeping gene and 16S rDNA gene ） , with another 7 type strains being used as reference strains. The results showed that these strains belonged to 1 genus, Komagataeibacter, and 3 known species, with another 14 strains being unknown species. This essay proposed that we should take the house-keeping genes, such as dnaK, rpoB, groEL gene, as the identification standard of these 93 strains, because the similarity values of 16S rDNA gene were to be above 99.56%. By pair-wise comparisons of dnaK, rpoB and groEL sequence for the 14 strains, the similarity values were to be above 99.0%, 98.8%, 99.2%, which suggested that the 14 isolates may belong to the same species. Currently, the strains labrary was classified to cover 9 known species and 1 proposed new species. These strains form a small but distinctive library, which laied a solid foundation on the genetic improvement of excellent strains, the conservation of BC-producing culture resource, and intensive study of bacterial cellulose.

关 键 词：椰纤果 细菌纤维素 菌种分离鉴定 菌种库建立 持家基因 16SrDNA序列分析 

分 类 号：S963.211[农业科学—水产养殖]