一株克服地黄连作障碍有益菌的鉴定及其LuxAB基因标记  

Identification and LuxAB Biochemical Mark of A Certain Bacterium Contributes to Settle Continuous Cropping Obstacles of Rehmannia glutinosa

作　　者：谭亚男[1,2] 马汇泉[1] 温学森[2] 刘舒[1] 

机构地区：[1]山东理工大学生命科学学院,山东淄博255049 [2]山东大学药学院,山东济南250100

出　　处：《安徽农业科学》2010年第17期8967-8969,共3页Journal of Anhui Agricultural Sciences

基　　金：国家"十一五"支撑计划项目(2006BA109B03);国家自然科学基金项目(30572326)

摘　　要：[目的]研究对克服地黄(Rehmannia glutinosa)连作障碍有益的43号菌的性质及其对地黄连作障碍的作用效果。[方法]利用细菌16S rDNA的保守序列进行比对,结合形态学观察、革兰氏染色和生理生化等特征对筛选得到的能够克服地黄连作障碍的43号菌进行鉴定;利用LuxAB发光酶基因对其进行标记,经抗性平板筛选,滴加葵醛在暗室进行检测,看是否得到了重组菌株;比较标记前后菌株对培养基中加入生地提取液的组培苗的作用效果。[结果]鉴定结果表明,43号菌为恶臭假单胞菌(Pseudomonas putida)。检测结果表明,得到重组菌株,命名为C-43。经过比较,标记前后菌株的生理生化特性基本没有变化,确定C-43对克服地黄连作障碍仍然有益。[结论]经过标记后的恶臭假单胞菌C-43号菌可用于后续的大田研究。[Objective]The research aimed to study the character of a certain bacterium which was benefit to settle continuous cropping obstacles of Rehmannia glutinosa and its effect.[Method]Combining with morphological observation,gram stain and comparison of physiological and biochemical characteristics,conservative sequence alignment of bacterial 16S rDNA was used to identify a certain bacterium 43.This strain was separated from the soil in which the Rehmannia glutinosa was planted and was benefit to settle continuous cropping obstacles of Rehmannia glutinosa.For a follow-up study in field experiment,LuxAB luminescent enzyme gene was used to mark this strain,then chose the recombinant strain by antibiotic selection plate and dropped kwai aldehyde in the panel to detect the characters in the dark room.Finally,the effects of bacterium 43 and C-43 on seedling cultivated in culture medium with Rehmannia glutinosa extract.[Result] Bacterium 43 belonged to Pseudomonas putida.And a recombination strain C-43 was obtained,which had the same basic physiological and biochemical characteristics compared to the original strain.Therefore the C-43 strain could also contribute to settle the continuous cropping obstacles of Rehmannia glutinosa.[Conclusion]The recombination strain C-43 can be used for follow-up study of the field experiments.

关 键 词：地黄 恶臭假单胞菌 生理生化特性 16S RDNA LuxAB发光酶基因 

分 类 号：S567.239[农业科学—中草药栽培]