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1、萬(wàn)此麈學(xué)報(bào)2013,22(1I):208—212ActaAgriculturaeBoreali—occidentalisSinicadoi:10.7606/j.issn.1004—1389.2013.11.036網(wǎng)絡(luò)出版時(shí)間:2013-11-22網(wǎng)絡(luò)出版地址:http://www.cnki.net/kcms/detail/61.1220.S.20131122.1659.037.html紅花三葉草中產(chǎn)苦馬豆素豆類絲核菌的分離與鑒定郭良,楊鳴琦,趙煜,羅頡,史嘉瑜,劉(西北農(nóng)林科技大學(xué)動(dòng)物醫(yī)學(xué)院,陜西楊凌712ioo)摘要為了確認(rèn)中國(guó)紅花三
2、葉草中是否存在產(chǎn)苦馬豆素(SW)的豆類絲核菌,用改良的真菌分離方法從植物樣品中分離豆類絲核菌。根據(jù)豆類絲核菌的形態(tài)特征對(duì)分離的真菌進(jìn)行初步篩選,然后對(duì)初步篩選所得真菌的5.8SrDNA—ITS序列進(jìn)行擴(kuò)增、測(cè)序并構(gòu)建系統(tǒng)發(fā)育樹以確定初步篩選真菌的種屬,經(jīng)薄層色譜和氣相色譜對(duì)豆類絲核菌菌絲體提取物進(jìn)行SW檢測(cè)。最終,篩選出一株符合豆類絲核菌形態(tài)特征的絲核菌屬真菌RL2012,薄層色譜和氣相色譜檢測(cè)到其菌絲體提取物中含有SW。綜上分析最終確認(rèn)紅花三葉草中存在產(chǎn)苦馬豆素的豆類絲核菌。關(guān)鍵詞豆類絲核菌;苦馬豆索;分離I鑒定中圖分類號(hào)$852.6
3、文獻(xiàn)標(biāo)志碼A文章編號(hào)1004—1389(2013)l1—0208—05IsolationandIdentificationofSwainsonine—producingRhizoctonialeguminicolafromRedCloverGU0Liang,YANGMingqi,ZHAOYu,LUOJie,SHIJiayuandLIUFang(CollegeofVeterinaryMedicine。NorthwestA&FUniversity,YanglingShaanxi712100,China)AbstractThestud芳ywa
4、stoconfirmwhetherredcloverwasinfectedbytheswainsonine—producingRhizoctonialeguminicolainChina.Rhizoctonialeguminicolawasisolatedfromredcloverbymodi—fledmethod.Firstly,Rhizoctonialeguminicolawasscreenedbyobservingthemorphologyofthefun—gi.Secondly,the5.8SrDNA—ITSsequenceof
5、thescreenedfungiwasamplifiedbyPCRandse—quenced。thephylogenetictreewasestablishedbasedonthe5.8SrDNA—ITSsequence.Thespeciesofscreenedfungiwasidentifiedbyanalyzingthephylogenetictree.Atlast,swainsoninewasdetectedinextractionofmyeeliumofRhizoctonialeguminicolabyTLCandGC.Ther
6、esultsshowedthataRhi—zoctoniafungusRL2O2whichhadmorphologicalcharacteristicsofRhizoctonialegurninicolawasiso一1ated;swainsoninewasfoundintheextractionofmyeeliumofRL2012byTLCandGC.Insummarywedeterminedthattheswainsonine—producingRhizoctonialeguminicolaexistintheredclover.K
7、eywordsRhizoctoniafeguminicola;Swainsonine;Isolation;Identification豆類絲核菌(Rhizoctonialeguminicola)為擔(dān)研究表明,引起流涎癥的主要物質(zhì)是豆類絲核菌子菌類(Basidiomycete),無(wú)孢菌群(Myceliasteril—的一種代謝產(chǎn)物,并命名為硫酰胺(slaframine,ia),絲核菌屬(Rhizoctonia)真菌,它可以侵染豆SF)。隨后,Schneider等報(bào)道從豆類絲核菌中科植物引起植物黑斑病,而動(dòng)物在食用豆類絲核分離出另一種代謝產(chǎn)
8、物苦馬豆素(swainsonine,菌污染的牧草后還會(huì)引起嚴(yán)重的流涎癥Ⅲ。早期SW),并發(fā)現(xiàn)其為引發(fā)流涎癥的另一種主要毒素。收稿日期:2013—05—13修回臼期I201306—04基金項(xiàng)目:國(guó)家自然科學(xué)基