| 土生空团菌(Cenococcum geophilum Fr.)的菌种鉴定及其遗传多样性的初步分析 |
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| 引用本文： | 陈立红,闫伟,徐燕.土生空团菌(Cenococcum geophilum Fr.)的菌种鉴定及其遗传多样性的初步分析[J].中国农业科学,2007,40(10):2214-2220. |
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| 作者姓名： | 陈立红 闫伟 徐燕 |
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| 作者单位： | 1. 内蒙古农业大学林学院,呼和浩特,010019;内蒙古农业大学农学院,呼和浩特,010019
2. 内蒙古农业大学林学院,呼和浩特,010019 |
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| 摘 要： | 【目的】鉴定外生菌根真菌土生空团菌（Cenococcum geophilum Fr.（Cg））菌种，分析土生空团菌的遗传多样性，探讨影响土生空团菌遗传分化的因素。【方法】采用形态特征结合PCR方法，从分离自6种寄主植物的27个中国菌株和来自法国的5个Cg菌株中鉴定出20个Cg菌株。利用PCR-限制性片段长度多态性（PCR-RFLP）和随机扩增片断多态性DNA（RAPD）两种分子标记方法对这20个Cg菌株进行遗传多样性分析。【结果】PCR-RFLP方法以通用引物ITS1和ITS4对20个Cg菌株的核糖体DNA转录间隔区（ITS）进行了PCR扩增，扩增产物用3种内切酶（EcoRⅠ、HinfⅠ和MboⅠ）酶切，酶切图谱显示菌株间有明显差异。RAPD方法用经过筛选的随机引物（5'-CGCACCGCAC-3'）对20个菌株的基因组DNA进行PCR扩增，扩增产物的片段大小在300～2 000 bp范围之间，共产生19个多态性位点。根据电泳图谱上DNA带的数目、位置及强度进行数值化，用聚类分析软件计算菌株间的遗传距离和遗传相似性，根据遗传距离构建20个菌株的系统聚类图。【结论】来源于不同寄主及地域的20株Cg有着较丰富的遗传多样性；地理环境和寄主对Cg遗传多样性的影响不大。
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| 关 键 词： | 外生菌根真菌 土生空团菌 限制性片段长度多态性 随机扩增片段多态性DNA 遗传多样性 |
| 收稿时间： | 2006-8-7 |
| 修稿时间： | 2006-07-28 |
Identification and Preliminary Analysis of Genetic Diversity of Cenococcum geophilum Fr. |
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| CHEN Li-hong,YAN Wei,XU Yan.Identification and Preliminary Analysis of Genetic Diversity of Cenococcum geophilum Fr.[J].Scientia Agricultura Sinica,2007,40(10):2214-2220. |
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| Authors: | CHEN Li-hong YAN Wei XU Yan |
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| Institution: | 1. College of Forestry, Inner Mongolia Agricultural University, Huhhot 010019; 2.College of Agronomy, Inner Mongolia Agricultural University, Huhhot 010019 |
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| Abstract: | Cenococcum geophilum Fr. (Cg) is an ectomycorrhizal fungus that has a wide host range and distribution. This mycobiont has been suggested to provide isolate-dependant drought protection to fine roots. It lacks asexual or sexual spores and froms abundant sclerotia that serve as resistant structures under environmental stress. The traditional classification method is very difficult to satisfy to the Cg classification and research of genetic diversity, so little is known about its genetic structure at fine scale. We obtained 27 Cg strains isolated from 6 hosts in China and 5 Cg strains introduced from France as our tested materials, and made identification based on morphological charactertics and PCR method. The genetic diversity of 20 Cg strains was estimated using RAPD and PCR-RFLP of the rDNA internal transcribed spacer (ITS) in this study. PCR-RFLP method used universal primers ITS1/ITS4 to amplified the rDNA ITS of mycelium, then amplified products were digested with HinfⅠand MboⅠ. The digested fragments of PCR products were run on 2% agarose gel, showed that there were obvious differences among strains. RAPD method used selected random primer (5'- CGCACCGCAC-3') to amplified genomic DNA of Cg RAPD produced 19 detectable and reliable DNA bands in the 300bp-2000bp size range. According to the number、position and strength of the DNA bands in agar gel, using PopGen Version 1.31 dendrogram analysis software calculated the genetic distance and genetic similarity among all tested strains. According to the genetic distance to constructed phylogenetic tree of 20 strains, the results indicated that a high degree of genetic diversity was observed among the Cg strains from the same or different hosts. |
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| Keywords: | Ectomycorrhizal fungi Cenococcum geophilum Fr RFLP RAPD Genetic diversity |
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