摘要
从分离出的知母内生细菌中筛选对人参6种常见病原菌和细辛3种常见病原菌有拮抗作用的菌株,为生物防治中药材病害提供基础。采用组织切片法和组织研磨法及菌丝生长速率法分离、筛选拮抗内生细菌。根据16S rDNA序列和系统发育树分析,对抑菌活性达到显著水平(P<0.05)的菌株进行菌种鉴定,并利用菌丝生长速率法检测活性菌株发酵液的抑菌率。通过正交试验优化该菌株的发酵条件。知母中分离出359株内生细菌,筛选出3株内生细菌BR10、BR11、BR26对这9种病原菌均有拮抗作用且抑菌活性较强,最高抑菌率达92.41%。16S rDNA序列系统发育分析结果显示BR11与多株水拉恩氏菌的亲缘关系最近,且处于系统发育树的同一分支,故将BR11菌株鉴定为水拉恩氏菌Rahnella aquatilis。确定了BR11菌株最佳培养基配方与培养条件:牛肉膏1.5%、葡萄糖1.5%、酵母膏0.5%、MgSO_(4)0.15%,初始pH值7.5,温度29℃,发酵时间72 h。知母内生细菌BR11是防治人参、细辛病害的优良候选菌株。
To screen antagonistic endophytic bacteria isolated from the plant of Anemarrhena asphodeloides,of which could inhibit six common diseases of Panax ginseng and three ones of Asarum sclerotium,to provide a base for biological control of some Chinese medicinal materials.The antagonistic endophytic bacteria were isolated and screened by tissue slicing,tissue grinding and hyphal growth rate.The strains with significant bacteriostatic activity(P<0.05)were identified according to 16S rDNA sequence and phylogenetic tree analysis,and the bacteriostatic rate of fermentation broth of active strains was detected by hyphal growth rate method.The fermentation conditions of the strain were optimized by orthogonal test.From the 359 endophytic bacteria isolated from the plant of A.asphodeloides three endophytic bacteria were screened,they are strains of BR10,BR11 and BR26,and all of them have had antagonistic effect and strong bacteriostatic activities to the nine pathogens,there was a highest bacteriostatic rate 92.41%of them.The results of phylogenetic analysis of 16S rDNA sequence showed that the BR11 had the closest relationship with many strains of Rahnella aquatilis,and was in the same branch of the phylogenetic tree,so the BR11 strain was identified as R.aquatilis.The optimum medium formula and culture conditions of BR11 strains were determined:beef paste 1.5%,glucose 1.5%,yeast extract 0.5%,MgSO_(4)0.15%,initial pH value 7.5,temperature 29℃,fermentation time 72 h.The endophytic bacteria BR11 from A.asphodeloides is an excellent candidate for the control of ginseng and Asarum diseases.
作者
孟千琪
王志清
刘桂英
马静
吕静
田义新
MENG Qianqi;WANG Zhiqing;LIU Guiying;MA Jing;LYU Jing;TIAN Yixin(College of Traditional Chinese Medicine,Jilin Agricultural University,Changchun 130118;Agricultural and Rural Bureau of Jiangyuan District,Baishan 134700,China)
出处
《东北农业科学》
2023年第1期45-49,66,共6页
Journal of Northeast Agricultural Sciences
基金
吉林省科技厅技术攻关项目(20190304017YY)
吉林省科技厅医药健康专项(20191102038YY)。
关键词
知母
内生细菌
病原菌
拮抗菌
发酵条件优化
Anemarrhena asphodeloides Bunge
Endophytic bacteria
Pathogenic bacteria
Antagonistic bacteria
Optimization of fermentation conditions
