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Plant Diversity ›› 2012, Vol. 34 ›› Issue (6): 614-622.DOI: 10.3724/SP.J.1143.2012.12140

• 研究论文 • 上一篇    下一篇

大型经济真菌的DNA条形码研究——以我国剧毒鹅膏为例

 蔡箐, 唐丽萍, 杨祝良   

  1. 中国科学院昆明植物研究所生物多样性与生物地理学院重点实验室,云南 昆明650201
  • 收稿日期:2012-11-05 出版日期:2012-12-25 发布日期:2012-11-09
  • 基金资助:

    国家科技部科技基础工作专项项目;国家高科技研究发展计划 (863计划) (2012AA021801);中国科学院大科学装置开放研究项目 (2009-LSF-GBOWS-01)

DNA Barcoding of Economically Important Mushrooms: A Case Study on Lethal Amanitas from China

 CA  Qing, TANG  Li-Ping, YANG  Zhu-Liang   

  1. Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, China
  • Received:2012-11-05 Online:2012-12-25 Published:2012-11-09

摘要:

鹅膏属(Amanita)部分物种为重要食用真菌,而另外部分物种则是剧毒的,在我国及其他许多国家,每年都有因误食剧毒鹅膏而导致中毒甚至死亡的事件发生。DNA条形码是用一段或几段短的DNA序列来对物种进行快速、准确鉴定的方法。本研究选取三个候选片段,即核糖体大亚基(nLSU)、内转录间隔区(ITS)和翻译延长因子1-α(tef1-α),使用真核生物通用引物,测试我国已知的7种剧毒鹅膏及2种易混的可食鹅膏,并将欧美分布的但与黄盖鹅膏(A.subjunquillea)亲缘关系密切的绿盖鹅膏(A.phalloides)纳入分析中。nLSU的PCR扩增和测序成功率均为100%,但种内和种间遗传变异偶有重叠。ITS的PCR扩增和测序成功率分别达到100%和85.7%,且具有高的种间变异和低的种内变异。tef1-α的PCR扩增和测序成功率分别达到85.7%和100%,种间和种内遗传分化均高于ITS和nLSU。三个片段的物种分辨率均较高,但与nLSU相比,ITS和tef1-α具有更为明显的barcode gap。鉴于ITS可能会成为真菌界的通用条码,故建议将ITS作为鹅膏属的核心条形码,tef1α和nLSU作为该属的辅助条形码。

关键词: DNA条形码, 食用野生蘑菇, 真菌, 毒蘑菇, 物种识别

Abstract:

Some species of the genus Amanita are economically important gourmet mushrooms, while others cause dramatic poisonings or even deaths every year in China and in many other countries. A DNA barcode is a short segment or a combination of short segments of DNA sequences that can distinguish species rapidly and accurately. To establish a standard DNA barcode for poisonous species of Amanita in China, three candidate markers, the large subunit nuclear ribosomal RNA (nLSU), the internal transcribed spacer (ITS), and the translation elongation factor 1alpha (tef1α) were tested using the eukaryotic general primers for their feasibility as barcodes to identify seven species of lethal fungi and two species of edible ones which can easily be confused with the lethal ones known from China. In addition, A.phalloides—a European and North American species closely related to one of the seven taxa, A.subjunquillea was also included. PCR amplification and sequencing success rate, intra and interspecific variation and rate of species identification were considered as main criteria for evaluation of the candidate DNA barcodes. Although the nLSU had high PCR and sequencing success rates (100% and 100% respectively), occasional overlapping occurred between the intra and interspecific variations. The PCR amplification and sequencing success rates of ITS were 100% and 85.7% respectively. ITS showed high sequence variation among species group and low variation within a given species. There was a relatively high PCR amplification and sequencing success rate for tef1α (85.7% and 100% respectively), and its intra and interspecific variation was higher than that of ITS or nLSU. All three candidate markers showed hight species resolution. ITS and tef1α had a more clearly defined barcode gap than nLSU. Our study showed that the tef1α and nLSU can be proposed as supplementary barcodes for the genus Amanita, while ITS can be used as a primary barcode marker considering that the ITS region may become a universal barcode marker for the fungal kingdom.

Key words: DNA barcoding, Edible wild mushrooms, Fungi, Poisonous mushrooms, Species recognition

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