GONG Na,MA Xiaoying,CHEN Xun,et al.Study on Classification of Oudemansiella raphanipes Based on HPCE and Multi Gene Combined Sequence Identification[J].Northern Horticulture,2022,(18):117-123.[doi:10.11937/bfyy.20221010]
基于HPCE和多基因联合序列鉴定卵孢长根菇的分类研究
- Title:
- Study on Classification of Oudemansiella raphanipes Based on HPCE and Multi Gene Combined Sequence Identification
- 文献标志码:
- A
- 摘要:
- 以5株卵孢长根菇菌株为试材，采用 ITS 和nLSU序列的测定并综合分析，结合GenBank中相关物种序列及菌株的形态学特征，进行卵孢长根菇的分子生物学鉴定，并且采用高效毛细管电泳检测，研究了卵孢长根菇的高效毛细管指纹图谱，通过主成分分析、相似度分析和聚类分析，对其进行鉴定和评价，以期探究新的快速准确的鉴定方法解决卵孢长根菇种类繁多，种内分类难度大，命名混乱等问题。结果表明：参试菌株总体亲缘关系较近，均为卵孢长根菇(Oudemansiella raphanipes)，Or2、Or5和Or1亲缘关系较近的菌株均不同程度的聚为一支，Or2、Or5和Or1相对于Or4、Or6亲缘关系更近，基于指纹图谱鉴别卵孢长根菇子实体形态的多样性，通过“W”形的三联差异峰作为鉴别卵孢长根菇不同子实体出菇表型的关键指示峰。卵孢长根菇具有丰富的物种多样性，采用HPCE和多基因联合序列的鉴定方法可有效区分不同形态种，为快速筛选优质卵孢长根菇菌株提供参考依据。
- Abstract:
- Five isolates of Oudemansiella raphanipes were used as the experimental materials.The molecular biological identification of Oudemansiella raphanipes was determined and comprehensive analysised by using ITS and nLSU sequence,combined with the sequences of related species in GenBank and morphological characteristics of the strains. In addition,high performance capillary electrophoresis was used to study the high performance capillary fingerprinting of Oudemansiella raphanipes,and the identification and evaluation were carried out by principal component analysis,similarity analysis and cluster analysis,in order to explore a new rapid and accurate identification method and to solve the problem of confusing nomenclature problems due to the large number of species of Oudemansiella raphanipes.The results showed that all test strains had close relationships,were all Oudemansiella raphanipes.Strains with close genetic relationships of Or2,Or5 and Or1 all clustered into one branch to varying degrees,Or2,Or5 and Or1 had closer genetic relationships than Or4 and Or6.The ‘W’ shaped triad differential peak was used as the key indicator peak for the identification of different fruiting bodies of Oudemansiella raphanipes.There was rich species diversity in Oudemansiella raphanipes.The identification method of HPCE and multi gene joint sequence could effectively distinguish different morphological species,which provided reference for rapid screening of high-quality Oudemansiella raphanipes strains.
参考文献/References:
[1]HAO Y J,ZHAO Q,WANG S X,et al.What is the radicate Oudemansiella cultivated in China?〖KG-*3〗[J].Phytotaxa,2016,286(1)：1-12.[2]张金霞,赵永昌.食用菌种质资源学[M].北京:科学出版社,2016.[3]TSANTFIZOS Y S,YANG X S,MCCLORY A.Studies on the biosynthesis of the fungal metabolite oudenone.Synthesis and enzymatic cyclization of analpha-diketone open-chainprecursor into oudenone in cuhures of Oudemansiella radicata[J].Journal of Organic Chemistry,1999,64（8）:6609-6614.[4]YANG Z L.Furthernotes on the Oudemansiella from southwestern China[J].Mycotaxon,2000,74:357-366.[5]PETERSEN R H,HUGHES K W.The xerula/oudemansiella complex (Agaricales)[J].Nova Hedwigia Beiheft,2010,137:1-625.[6]YANG Z L,ZHANG L F,MUELLER G M,et al.A newsystematic arrangement of the genus Oudemansiellas.str.(Physalacriaceae,Agaricales)[J].Mycosystema,2009,28(1):1-13.[7]BEAN S R,LOOKHART G L.High-performance capillary electrophoresis of meat,dairy,and cereal proteins[J].Electrophoresis,2001,22(19):4207-4215.[8]KAISOON O,SIRIAMORNPUN S,MEESO N.Distinction between cereal genotypes based on the protein and DNA composition of the grain by capillary electrophoresis[J].World Applied Sciences Journal,2008(4):384-395.[9]ZELLER F J,YAN Y M,YU J Z,et al.Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers[J].Electrophoresis,2003(24):1429-1436.[10]马晓颖,贾东贝,李冰宇,等.利用高效毛细管电泳测定真菌代谢产物吲哚乙酸含量[J].江苏农业科学,2011,39(3):439-441.[11]吴怡.灵芝属漆酶高产菌株的筛选及其所产漆酶的纯化和性质分析[D].北京:北京林业大学,2020.[12]马晓颖,肖军,杨涛,等.高效毛细管电泳法测定桑黄菌丝体提取物中腺苷含量的研究[J].园艺与种苗,2020,40(5):27-29.[13]马晓颖,杨镇,宋艳雨,等.高效毛细管电泳-二极管阵列检测法测定微生物代谢产物中的植物激素含量[J].江苏农业科学,2017,45(8):169-172.[14]袁晓艳,张书山,王明阳,等.贵州产灵芝HPLC指纹图谱研究[J].遵义医科大学学报,2019,42(6):641-645.[15]马晓颖,吕立涛,曹君,等.羊肚菌毛细管电泳指纹图谱的研究[J].北方园艺,2019(12):131-138.[16]VAL R H,MIADLIKOWSKA J,KAUFF F,et al.Phylogenetic comparison of protein-coding versus ribosomal RNA-coding sequence data：A case study of the Lecanoromycetes (Ascomycota)[J].Molecular Phylogenetics and Evolution,2007,44(1):412-426.[17]LIU Y J,WHELEN S,HALL B D.Phylogenetic relationships among ascomycetes：Evidence from an RNA polymerse Ⅱ subunit[J].Molecular Biology and Evolution,1999,16(12):1799-1808.[18]P LDMAA K.Tropical species of Cladobotryum and Hypomyces producing red pigments[J].Studies in Mycology,2011,68：1-34.[19]刘国丽,李红,龚娜,等.基于形态特征和多基因联合序列鉴定卵孢长根菇蛛网病病原菌[J].食用菌学报,2020,27(3)：30-36.[20]苏春丽,唐传红,张劲松,等.基于rDNA ITS序列探讨中国栽培灵芝菌株的亲缘关系[J].微生物学报,2007(1):11-16.[21]谢占玲,田飞,余静,等.基于LSU 和 ITS 的青藏高原黄绿卷毛菇种群遗传多样性分析[J].菌物学报,2015,34(1)：26-37.[22]RANS T D,SHEFFERSON R P.Evolutionary studies of ectomycorrhizal fungi：Recent advances and future directions[J].Can J Bot,2004,82（8）：1122-1132.[23]王卫东,高翔,赵丹阳.高分子量麦谷蛋白亚基 HPCE 高效分离及图谱鉴定[J].作物学报,2018,44(7)：966-976.[24]BEAN S R,LOOKHART G L.Recent developments in high-performance capillary electrophoresis of cereal proteins[J].Electrophoresis,2001，22(8)：1503-1509.[25]ARYANI S,MOHAMAD R，LATIFAHK D.Discrimination of red and white rice bran from indonesia using HPLC fingerprint analysis combined with chemometrics[J].Food Chemistry,2017,221(15):1717-1722.[26]常冰.不同桂花品种GC/MS和HPLC指纹图谱分析[D].郑州:河南大学,2019.[27]孟虎,孙国琴,睢韡,等.ISSR技术在食用菌研究上的应用[J].北方园艺,2016(5)：207-210.
相似文献/References:
[1]程立君,罗家刚,胡志芳,等.四个天麻品种种植效果的对比分析[J].北方园艺,2016,40(20):156.[doi:10.11937/bfyy.201620040]
CHENG Lijun,LUO Jiagang,HU Zhifang,et al.Planting Effect Comparison of Four Varieties of Gastrodia elata[J].Northern Horticulture,2016,40(18):156.[doi:10.11937/bfyy.201620040]
[2]赵思思,刘兴菊,李雪雁,等.基于成熟期的梨品种遗传多样性SSR分析[J].北方园艺,2017,41(16):43.[doi:10.11937/bfyy.20170631]
ZHAO Sisi,LIU Xingju,LI Xueyan,et al.Genetic Diversity of SSR Analysis of Pear Cultivars Based on Maturity[J].Northern Horticulture,2017,41(18):43.[doi:10.11937/bfyy.20170631]
备注/Memo
第一作者简介：龚娜(1982-)，女，硕士，副研究员，现主要从事食药用菌育种及栽培生理等研究工作。E-mail：doll52133@163.com.责任作者：肖军（1974-），女，硕士，研究员，现主要从事食药用菌育种等研究工作。E-mail：214081450@qq.com.基金项目：揭榜挂帅科技攻关专项资助项目(2021JH1/10400035)；辽宁省农业科学院院长基金资助项目(2022QN2303)；辽宁省科技特派活动专项计划资助项目(2021JH5/10400035)；沈阳市科技特团专项资助项目(21-116-3-23)；国家食用菌产业技术体系资助项目(CARS-20)。收稿日期：2022-03-18