基于DNA条形码技术鉴别有毒鹅膏菌属物种  

DNA Barcoding for Identification of Toxic Amanita Species

作　　者：白文明 邢冉冉 陈丽萍 彭涛[2] 雷红涛[1] 陈颖[2] BAI Wenming;XING Ranran;CHEN Liping;PENG Tao;LEI Hongtao;CHEN Ying(College of Food Science,South China Agricultural University,Guangzhou 510642,China;Chinese Academy of Inspection and Quarantine,Beijing 100176,China;Inspection and Quarantine Technical Center,Kunming Customs District P.R.China,Kunming 650051,China)

机构地区：[1]华南农业大学食品学院,广东广州510642 [2]中国检验检疫科学研究院,北京100176 [3]中华人民共和国昆明海关检验检疫技术中心,云南昆明650051

出　　处：《食品科学》2021年第4期278-286,共9页Food Science

基　　金：“十三五”国家重点研发计划重点专项(2017YFF0211301)。

摘　　要：收集27个鹅膏菌属物种共38份样本,提取样品基因组DNA,应用通用引物扩增其内转录间隔区(internal transcribed spacer,ITS)、核糖体大亚基(large ribosomal subunit,LSU)、RNA聚合酶的第二大亚基(the second largest subunit of RNA polymerase II,RPB2)、β-微管蛋白(β-tubulin)基因序列并进行Sanger双向测序,将得到的序列进行校对拼接后与NCBI的GenBank数据库中的参考序列进行比对鉴别物种来源;计算物种的种内、种间Kimura-2-Parameter(K2P)遗传距离并构建系统发育树。结果表明,β-tubulin、ITS基因序列鉴别能力优于RPB2、LSU基因序列,可将β-tubulin与ITS两者联合用于鹅膏菌属的物种鉴别,为有毒蘑菇诱发的食源性中毒风险进行预警。β-tubulin基因序列长度较LSU、ITS、RPB2等基因序列短,适合对深加工的蘑菇制品以及误食毒蘑菇后的呕吐物进行分析,可作为鹅膏菌属中毒事件中物种鉴定及溯源的优选条形码。In this study,we collected a total of 38 samples of 27 Amanita species and extracted their genomic DNA.Universal primers were used to amplify the internal transcribed spacer(ITS),large ribosomal subunit(LSU),the secondlargest subunit of RNA polymerase II(RPB2),and the β-tubulin gene sequences.Sanger bidirectional sequences were obtained,proofread and then submitted to the NCBI GenBank for sequence alignment to identify the species.We calculated the intra-species and inter-species Kimura-2-Parameter(K2P)genetic distance and constructed the phylogenetic tree.The results indicated that β-tubulin and ITS were more suitable than RPB2 and LSU for use in the identification of Amanita species.The combined use ofβ-tubulin and ITS could be recommended to identify Amanita species,providing early warning of foodborne poisoning caused by poisonous mushrooms.β-tubulin was shorter than LSU,ITS,and RPB2,being suitable for use in the analysis of highly-processed mushroom products and vomits after eating poisonous mushrooms by mistake.Thus,β-tubulin can be used as the optimal barcode to identify and trace Amanita species causing mushroom poisoning.

关 键 词：鹅膏菌属 DNA条形码 物种鉴别 

分 类 号：Q939.5[生物学—微生物学]