红花龙胆内生真菌多样性分析及抑菌活性菌株筛选  

Diversity Analysis of Endophytic Fungi Isolated from Gentiana rhodantha and Screening for Biocontrol Stains

作　　者：张晓勇 李树江 严凯 向红 杨友联 Zhang Xiaoyong;Li Shujiang;Yan Kai;Xiang Hong;Yang Youlian(School of Biological Science and Technology,Liupanshui Normal University,Liupanshui,553004)

机构地区：[1]六盘水师范学院生物科学与技术学院,六盘水553004

出　　处：《分子植物育种》2021年第14期4714-4725,共12页Molecular Plant Breeding

基　　金：贵州省教育厅青年科技人才成长项目(黔教合KY字〔2018〕381);贵州省特色重点实验室建设项目(黔教合KY字〔2017〕012);六盘水师范学院教务处教学改革项目(LPSSYjxtd201802);六盘水市科技局基金项目(52020-2017-02-10)共同资助。

摘　　要：为了明确民族传统中药材红花龙胆(Gentiana rhodantha Franch.ex Hemsl)内生真菌群落结构及功能,本研究采用组织块分离法,对贵州省六盘水地区的野生红花龙胆内生菌进行分析,结合形态特征和ITS序列对分离的菌株进行鉴定,并阐述内生菌多样性;采用平板对峙法筛选能显著抑制红花龙胆菌核病菌(Sclerotinia sclerotionrum)生长的内生菌株。结果表明:从六盘水地区钟山凤凰、水城南开、水城杨梅和六枝新场等4个点的480个野生红花龙胆组织块中分离获得294株内生真菌,隶属32个属,总定殖率和总分离率分别为49.38%和61.25%,内生真菌多样性非常丰富。从总体分离频率分布来看,镰刀菌属(Fusarium)(9.86%)和刺盘孢属(Colletotrichum)(8.84%)为优势菌群。多样性分析结果显示,六枝新场的红花龙胆内生菌多样性程度最高,在不同部位中茎秆的内生真菌多样性大于叶片和根部。通过与丝核菌进行对峙培养筛选出抑制率超过50%的有23株内生真菌,其中葡萄座腔属的R022、R021、拟茎点霉属的S002、刺盘孢属的S028以及镰刀菌属的L031等5株内生菌对红花龙胆菌核病菌菌丝抑制率均超过70%,具有较大的生防潜力。本研究初步探明红花龙胆内生真菌多样性,并为红花龙胆菌核病生物防治研究奠定基础。In order to clarify the composition of endophytic fungi in Gentiana rhodantha in colonization tissues with four Liupanshui sampling sites(Fenghuang Zhongshan,Nankai Shuicheng,Yangmei Shuicheng and Xinchang Liuzhi),strains were isolated with health leaf,stem and root and identification based on morphological characteristics and ITS sequence analysis.As well as screening for biocontrol stains by the dural culture method on PDA plates with Sclerotinia sclerotionrum.The results showed that G.rhodantha concealed rich fungal diversity,the colonization rate and isolation rate of endophytic fungi reached 49.38%and 61.25%respectively.294 strains of endophytic fungi were obtained from 480 explant tissue segments of G.rhodantha and belonging to 2 phylums,5 classes,14 orders and 32 genera of which Fusarium and Colletotrichum were comparatively advantageous taxas in endophytic fungi of G.rhodantha,and the species contained occupied 9.86%and 8.84%of the total endophytes respectively.According to the calculation results,Xinchang Liuzhi in four sites and stem in three tissues showed optimum degrees of diversity,evenness and dominance respectively.23 strains of endophytic fungi with inhibitory activities against S.sclerotionrum by the dural culture method.Among them,the inhibition rate of Botryosphaeria sp.R022,R021,Phomopsis sp.S002,Colletotrichum sp.S028 and Fusarium sp.L031 were higher than 70%and showed the best inhibitory potential.This study demonstrated the diversity of endophytic fungi in Gentiana sclerotiorum,and laid a foundation for the biological control of Gentiana sclerotiorum.

关 键 词：红花龙胆(Gentiana rhodantha) 内生真菌 多样性 核盘菌 拮抗菌株 

分 类 号：S567.239[农业科学—中草药栽培]