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1、微生物学通报OCT20,2010,37(10):1467−1474MicrobiologyChina©2010byInstituteofMicrobiology,CAStongbao@im.ac.cn研究报告怀地黄活性内生菌的分离鉴定及抗菌抗肿瘤活性*杨清香谢永生张昊李学梅(河南师范大学生命科学学院河南新乡453007)摘要:利用平板分离法从怀地黄(RehmanniaglutinosaLibosch)中分离出内生菌共130株,包括67株细菌、50株真菌和13株放线菌。利用大肠杆菌、金黄色葡萄球菌和黑曲霉作为供试菌株,通过对峙试验和平板点接法筛选出8株对3种供试菌株均具有较强拮抗作用的内生菌。
2、经菌种形态观察、生理生化实验以及16SrRNA序列测定,这8株活性内生菌均为假单胞菌属的细菌,并分别与Pseudomonasfluorescens、Pseudomonasthivervalensis、Pseudomonaschlororaphis、Pseudomonaskoreensis和一个未定种具有最高相似性。对这8株活性内生菌进行液体发酵、不同有机溶剂抽提和抗菌抗肿瘤研究,结果表明它们的乙醇抽提物和乙酸乙酯抽提物均不同程度地对3种供试菌株和食管癌细胞系Ec9706具有抑制作用。其中2-2号菌株(Pseudomonaschlororaphis)抗菌抗肿瘤作用最强,具有重要的开发价值。关键
3、词:内生菌,怀地黄,抗菌,抗肿瘤Isolation,IdentificationandAntagonismonMicroorganismsandCancerCellsbyActiveEndophytesfromRehmanniaglutinosaLibosch*YANGQing-XiangXIEYong-ShengZHANGHaoLIXue-Mei(CollegeofLifeSciences,HenanNormalUniversity,Xinxiang,Henan453007,China)Abstract:Usingthemethodsofplatecultivation,130strain
4、sofendophyteswereisolatedfromReh-manniaglutinosaLibosch,including67bacteria,50fungiand13actinomycetes.Fromthem,8isolatesofbacteriawereselectedduetotheirhighandwideantagonisticactivitiestothetestedmicrobialstrains,Escherichiacoli,StaphalocolcusaureusandAspergillusnigerontheplateconfrontation.The8act
5、ivestrainsofbacteriawereclassifiedtogenusofPseudomonasandsharedthehighestidentitiesre-spectivelywith5speciesincludingPseudomonasfluorescens,Pseudomonasthivervalensi,Pseudomo-naschlororaphis,Pseudomonaskoreensisandoneunidentifiedaccordingtotheirmorphological,phy-siologicalandchemicalcharacteristicsa
6、nd16SrRNAsequences.Activeproductswereobtainedfromthe8strainsthroughliquidfermentationandfurtherextractionbydifferentorganicsolvents.Activitystudiesindicatedthattheextractionproductsfromallthe8strainsusingethylacetateorethanolexhi-bitedsignificantinhibitiononthe3testedtypesofmicroorganismsandesophag
7、ealcancercellsEc9706.基金项目：国家自然科学基金项目(No.20677014)*通讯作者：Tel:86-373-3325528;:yangqx66@163.com收稿日期：2010-04-06;接受日期：2010-06-221468微生物学通报2010,Vol.37,No.10Amongthem,strain2-2hadthehighestactivitiesofbothanti-micr
