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1、微生物學(xué)通報(bào)OCT20,2010,37(10):1467?1474MicrobiologyChina?2010byInstituteofMicrobiology,CAStongbao@im.ac.cn研究報(bào)告懷地黃活性內(nèi)生菌的分離鑒定及抗菌抗腫瘤活性*楊清香謝永生張昊李學(xué)梅(河南師范大學(xué)生命科學(xué)學(xué)院河南新鄉(xiāng)453007)摘要:利用平板分離法從懷地黃(RehmanniaglutinosaLibosch)中分離出內(nèi)生菌共130株,包括67株細(xì)菌、50株真菌和13株放線菌。利用大腸桿菌、金黃色葡萄球菌和黑曲霉作為供試菌株,通過(guò)對(duì)峙試驗(yàn)和平板點(diǎn)接法篩選出8株對(duì)3種供試菌株均具有較強(qiáng)拮抗作用的內(nèi)生菌。
2、經(jīng)菌種形態(tài)觀察、生理生化實(shí)驗(yàn)以及16SrRNA序列測(cè)定,這8株活性內(nèi)生菌均為假單胞菌屬的細(xì)菌,并分別與Pseudomonasfluorescens、Pseudomonasthivervalensis、Pseudomonaschlororaphis、Pseudomonaskoreensis和一個(gè)未定種具有最高相似性。對(duì)這8株活性內(nèi)生菌進(jìn)行液體發(fā)酵、不同有機(jī)溶劑抽提和抗菌抗腫瘤研究,結(jié)果表明它們的乙醇抽提物和乙酸乙酯抽提物均不同程度地對(duì)3種供試菌株和食管癌細(xì)胞系Ec9706具有抑制作用。其中2-2號(hào)菌株(Pseudomonaschlororaphis)抗菌抗腫瘤作用最強(qiáng),具有重要的開發(fā)價(jià)值。關(guān)鍵
3、詞:內(nèi)生菌,懷地黃,抗菌,抗腫瘤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.基金項(xiàng)目：國(guó)家自然科學(xué)基金項(xiàng)目(No.20677014)*通訊作者：Tel:86-373-3325528;:yangqx66@163.com收稿日期：2010-04-06;接受日期：2010-06-221468微生物學(xué)通報(bào)2010,Vol.37,No.10Amongthem,strain2-2hadthehighestactivitiesofbothanti-micr
