本文主要研究内容
作者迟惠荣（2019）在《多花黄精叶枯病病原菌鉴定及贝莱斯芽胞杆菌防病促生效果研究》一文中研究指出：多花黄精为百合科黄精属植物,被《中国药典》(2015年版)收录,是一味药食同源的中药材。截至2018年,人工栽培已成为黄精主要生产模式,为此栽培过程中常受到多种病原菌侵害,如叶斑病、炭疽病等。在浙江省黄精种植基地中,发现一种发病比较严重的病害,该病的发病症状为在叶片顶端先形成病斑,后不断扩大,最终整个植株叶片枯死,为区分此种症状,起名为多花黄精叶枯病,该病害严重影响多花黄精根茎的生长和产量。为此,本文对多花黄精叶枯病的病原、多花黄精的内生菌、生防菌的防病效果及其拮抗机制和促生机理进行了研究,具体结果如下:1、对多花黄精的感病植株进行病原菌的分离、鉴定,分离得到5株真菌,对其进行致病性测定,发现只有菌株PC-4具有致病性。根据形态学、rDNA-ITS和TEF-1α序列同源性分析,对该病害的致病菌种进行鉴定,最终确定导致多花黄精叶枯病的病原菌为尖孢镰刀菌Fusarium oxysporum。2、从多花黄精根茎中分离获得11株内生细菌,经鉴定分别属于3个属,其中芽胞杆菌属为多花黄精内生细菌的优势菌属,采用平板对峙法筛选出一株对尖孢镰刀菌F.oxysporumPC-4具有抗菌效果的内生拮抗菌ZJU-3,通过形态学观察、生理生化测定、16S rDNA和gyrB基因序列同源性分析,最终鉴定菌株ZJU-3鉴定为贝莱斯芽胞杆菌Bacillusvelezensis。3、盆栽实验测定B.velezensisZJU-3的生防效果,发现OD600=0.5生防菌液的病情指数为39.5%,而只接种病原菌的对照组的病情指数为70.8%,防病效果为44.2%,可见B.velezensisZJU-3可有效降低发病率。4、多花黄精和滇黄精组培试验中,发现B.velezensB.ZJU-3发酵液均可以对株高、鲜重有促生作用,对叶片中叶绿素含量无影响。温室栽培试验结果发现该菌株对多花黄精具有显著的促生效果,根长和单株根数明显高于对照组。对发酵液中激素种类进行利用液相色谱-质谱联用(LC-MS)进行分析,发现B.velezensis ZJU-3产生多种植物内源激素如IAA、kinetin、zeatin、GA3,其浓度依次为8.29 ng/mL、21.34 ng/mL、0.02 ng/mL、0.07 ng/mL。5、利用浓度梯度法对菌株进行利福平的抗性诱导,筛选出保持抗菌活性的抗利福平的突变体Rif-B.velezensis ZJU-3,其对利福平的抗性达到150 μg/mL。通过叶片接种、组织分离,发现叶片内Rif-B.velezensis ZJU-3菌株数量为2×105个/g。6、菌丝生长速率测定结果显示,拮抗菌ZJU-3无菌发酵液对F.oxysporum PC-4菌丝生长均有抑制作用,随着培养基中发酵液添加比例的增加,抑菌效果增强,当加入5%发酵滤液时,菌落直径为3.78cm,对照为4.87cm,抑制率为22.3%。对B.velezensisZJU-3发酵液抗菌活性稳定性分析,发现贝莱斯芽胞杆菌B.velezensisZJU-3发酵液抗菌活性不受温度、pH、蛋白酶K和金属盐离子影响。7、抑菌谱测定试验发现拮抗菌ZJU-3对铁皮石斛炭疽病菌(Colletotrichum gloeosporioides)、大豆 炭疽病(C.truncatum)、小麦赤霉病菌(F.graminearum)、西红花茄病镰刀菌(F.solani)、棉花立枯丝核病菌(Rhizoctoniasolani)、西瓜枯萎病菌(F.oxyporum f.sp.niveum)的菌丝都有较强的抑制作用,抑菌效果分别为71.8%、79%、55%、58.4%、75.1%和66.4%,且部分病菌菌丝缩短变粗、扭曲、变黑、表面出现褶皱,产孢量明显减少,可说明该菌株具有一定的广谱抗性。8、抑菌物质检测发现,B.velezensis ZJU-3能够产生嗜铁素、蛋白酶、纤维素酶和β-葡聚糖酶,不分泌几丁质酶。根据脂肽类抗生素合成相关基因序列(fenA、sfp、ituA)对目标菌株进行PCR扩增,发现贝莱斯芽胞杆菌B.velezensB.ZJU-3的基因组中可能存在泛革素、伊枯草菌素和表面活性素的代谢合成操纵子序列。MALDI-TOF-MS检测结果表明生防菌B.velezensis能够产生丰富的脂肽类抗生素,包括泛革素、表面活性素和伊枯草菌素三个家族。利用盐酸沉淀和甲醇抽提法获得了菌株ZJU-3发酵液中脂肽粗提物,确定该脂肽粗提物具有明显抑制尖孢镰刀菌菌丝生长的能力,抑制率达到51.6%。
Abstract
duo hua huang jing wei bai ge ke huang jing shu zhi wu ,bei 《zhong guo yao dian 》(2015nian ban )shou lu ,shi yi wei yao shi tong yuan de zhong yao cai 。jie zhi 2018nian ,ren gong zai pei yi cheng wei huang jing zhu yao sheng chan mo shi ,wei ci zai pei guo cheng zhong chang shou dao duo chong bing yuan jun qin hai ,ru xie ban bing 、tan ju bing deng 。zai zhe jiang sheng huang jing chong zhi ji de zhong ,fa xian yi chong fa bing bi jiao yan chong de bing hai ,gai bing de fa bing zheng zhuang wei zai xie pian ding duan xian xing cheng bing ban ,hou bu duan kuo da ,zui zhong zheng ge zhi zhu xie pian ku si ,wei ou fen ci chong zheng zhuang ,qi ming wei duo hua huang jing xie ku bing ,gai bing hai yan chong ying xiang duo hua huang jing gen jing de sheng chang he chan liang 。wei ci ,ben wen dui duo hua huang jing xie ku bing de bing yuan 、duo hua huang jing de nei sheng jun 、sheng fang jun de fang bing xiao guo ji ji jie kang ji zhi he cu sheng ji li jin hang le yan jiu ,ju ti jie guo ru xia :1、dui duo hua huang jing de gan bing zhi zhu jin hang bing yuan jun de fen li 、jian ding ,fen li de dao 5zhu zhen jun ,dui ji jin hang zhi bing xing ce ding ,fa xian zhi you jun zhu PC-4ju you zhi bing xing 。gen ju xing tai xue 、rDNA-ITShe TEF-1αxu lie tong yuan xing fen xi ,dui gai bing hai de zhi bing jun chong jin hang jian ding ,zui zhong que ding dao zhi duo hua huang jing xie ku bing de bing yuan jun wei jian bao lian dao jun Fusarium oxysporum。2、cong duo hua huang jing gen jing zhong fen li huo de 11zhu nei sheng xi jun ,jing jian ding fen bie shu yu 3ge shu ,ji zhong ya bao gan jun shu wei duo hua huang jing nei sheng xi jun de you shi jun shu ,cai yong ping ban dui shi fa shai shua chu yi zhu dui jian bao lian dao jun F.oxysporumPC-4ju you kang jun xiao guo de nei sheng jie kang jun ZJU-3,tong guo xing tai xue guan cha 、sheng li sheng hua ce ding 、16S rDNAhe gyrBji yin xu lie tong yuan xing fen xi ,zui zhong jian ding jun zhu ZJU-3jian ding wei bei lai si ya bao gan jun Bacillusvelezensis。3、pen zai shi yan ce ding B.velezensisZJU-3de sheng fang xiao guo ,fa xian OD600=0.5sheng fang jun ye de bing qing zhi shu wei 39.5%,er zhi jie chong bing yuan jun de dui zhao zu de bing qing zhi shu wei 70.8%,fang bing xiao guo wei 44.2%,ke jian B.velezensisZJU-3ke you xiao jiang di fa bing lv 。4、duo hua huang jing he dian huang jing zu pei shi yan zhong ,fa xian B.velezensB.ZJU-3fa jiao ye jun ke yi dui zhu gao 、xian chong you cu sheng zuo yong ,dui xie pian zhong xie lu su han liang mo ying xiang 。wen shi zai pei shi yan jie guo fa xian gai jun zhu dui duo hua huang jing ju you xian zhe de cu sheng xiao guo ,gen chang he chan zhu gen shu ming xian gao yu dui zhao zu 。dui fa jiao ye zhong ji su chong lei jin hang li yong ye xiang se pu -zhi pu lian yong (LC-MS)jin hang fen xi ,fa xian B.velezensis ZJU-3chan sheng duo chong zhi wu nei yuan ji su ru IAA、kinetin、zeatin、GA3,ji nong du yi ci wei 8.29 ng/mL、21.34 ng/mL、0.02 ng/mL、0.07 ng/mL。5、li yong nong du ti du fa dui jun zhu jin hang li fu ping de kang xing you dao ,shai shua chu bao chi kang jun huo xing de kang li fu ping de tu bian ti Rif-B.velezensis ZJU-3,ji dui li fu ping de kang xing da dao 150 μg/mL。tong guo xie pian jie chong 、zu zhi fen li ,fa xian xie pian nei Rif-B.velezensis ZJU-3jun zhu shu liang wei 2×105ge /g。6、jun si sheng chang su lv ce ding jie guo xian shi ,jie kang jun ZJU-3mo jun fa jiao ye dui F.oxysporum PC-4jun si sheng chang jun you yi zhi zuo yong ,sui zhao pei yang ji zhong fa jiao ye tian jia bi li de zeng jia ,yi jun xiao guo zeng jiang ,dang jia ru 5%fa jiao lv ye shi ,jun la zhi jing wei 3.78cm,dui zhao wei 4.87cm,yi zhi lv wei 22.3%。dui B.velezensisZJU-3fa jiao ye kang jun huo xing wen ding xing fen xi ,fa xian bei lai si ya bao gan jun B.velezensisZJU-3fa jiao ye kang jun huo xing bu shou wen du 、pH、dan bai mei Khe jin shu yan li zi ying xiang 。7、yi jun pu ce ding shi yan fa xian jie kang jun ZJU-3dui tie pi dan hu tan ju bing jun (Colletotrichum gloeosporioides)、da dou tan ju bing (C.truncatum)、xiao mai chi mei bing jun (F.graminearum)、xi gong hua jia bing lian dao jun (F.solani)、mian hua li ku si he bing jun (Rhizoctoniasolani)、xi gua ku wei bing jun (F.oxyporum f.sp.niveum)de jun si dou you jiao jiang de yi zhi zuo yong ,yi jun xiao guo fen bie wei 71.8%、79%、55%、58.4%、75.1%he 66.4%,ju bu fen bing jun jun si su duan bian cu 、niu qu 、bian hei 、biao mian chu xian zhe zhou ,chan bao liang ming xian jian shao ,ke shui ming gai jun zhu ju you yi ding de an pu kang xing 。8、yi jun wu zhi jian ce fa xian ,B.velezensis ZJU-3neng gou chan sheng shi tie su 、dan bai mei 、qian wei su mei he β-pu ju tang mei ,bu fen bi ji ding zhi mei 。gen ju zhi tai lei kang sheng su ge cheng xiang guan ji yin xu lie (fenA、sfp、ituA)dui mu biao jun zhu jin hang PCRkuo zeng ,fa xian bei lai si ya bao gan jun B.velezensB.ZJU-3de ji yin zu zhong ke neng cun zai fan ge su 、yi ku cao jun su he biao mian huo xing su de dai xie ge cheng cao zong zi xu lie 。MALDI-TOF-MSjian ce jie guo biao ming sheng fang jun B.velezensisneng gou chan sheng feng fu de zhi tai lei kang sheng su ,bao gua fan ge su 、biao mian huo xing su he yi ku cao jun su san ge jia zu 。li yong yan suan chen dian he jia chun chou di fa huo de le jun zhu ZJU-3fa jiao ye zhong zhi tai cu di wu ,que ding gai zhi tai cu di wu ju you ming xian yi zhi jian bao lian dao jun jun si sheng chang de neng li ,yi zhi lv da dao 51.6%。
论文参考文献
论文详细介绍
论文作者分别是来自浙江大学的迟惠荣，发表于刊物浙江大学2019-05-20论文，是一篇关于多花黄精论文,尖孢镰刀菌论文,内生细菌论文,贝莱斯芽胞杆菌论文,生防作用论文,促生作用论文,浙江大学2019-05-20论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自浙江大学2019-05-20论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：多花黄精论文; 尖孢镰刀菌论文; 内生细菌论文; 贝莱斯芽胞杆菌论文; 生防作用论文; 促生作用论文; 浙江大学2019-05-20论文;