利用RAPD技术对木耳属菌株进行分类鉴定的研究
阎培生 罗信昌 周启
摘 要:利用RAPD技术对木耳属不同种和种内不同菌株进行了分子鉴定。结果表明,在供试的26个随机引物中,有10个引物可扩增出清晰、稳定的DNA带型,其中引物S4和S6与供试木耳属菌株的基因组DNA结合位点少而保守,扩增图谱具有种的特异性,可用于种的快速准确鉴定;其综随机引物的扩增DNA多态性较强,可用于种内不同菌株的鉴定;聚类分析表明,在75%相似水平下,可将供试的木耳属菌株分成四大类,其中有三大类各代表一个形态鉴定的种。研究结果表明了RAPD技术可有效地用于木耳属种或菌株的快速准确鉴定。
关键词:随机扩增多态性DNA,分子鉴定,木耳属,聚类分析
中图分类号:Q936, 文献标识码:A.
文章编号:1007-3515(1999)03-0029-0033
CLASSIFICATION AND IDENTIFICATION OF
SPECIES AND STRAINS IN AURICULARIA
BY USING RAPD
YAN Pei-Sheng,LUO Xin-Chang,ZHOU Qi
(Laboratories of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070)
YAN Pei-Sheng
( Lab of Applied Mycology, Department of Plant Protection, Laiyang Agricultural College, Laiyang, Shandong, 265200)
ABSTRACT: RAPD technique was used to characterize and classify the species and strains of the genus Auricularia. The results showed that the annealing sites between random primer S4 or S6 and genomic DNA of different species were rare and conserved. The amplification patterns were species?specific. The two primers could be used in species identification directly; other tested primers could generate more polymorphic DNA and could be used in strain identification. Cluster analysis results showed that at 75% similarity level, all strains could be grouped into four clusters. Of the four clusters, three represented one morphological species respectively. RAPD may provide a useful and rapid tool for identifying species and strains of Auricularia.
KEY WORDS: RAPD, Molecular taxonomy, Auricularia, Cluster analysis.
国家自然科学基金资助项目
阎培生(华中农业大学农业部农业微生物重点实验室 武汉 430070)(莱阳农学院植保系应用真菌研究室 莱阳 265200)
罗信昌(华中农业大学农业部农业微生物重点实验室 武汉 430070)
周启(华中农业大学农业部农业微生物重点实验室 武汉 430070)
[参 考 文 献]
1,刘学敏,惠东威,张明厚等,1997.大豆灰斑病菌生理小种的RAPD标记.菌物系统,16(2):128~133
2,张引芳,1994.Studies on the differentiation of Lentinula edodes by RAPD assay. 食用菌学报,1(1):21~27
3,李丽嘉,1984.木耳属二新种.真菌学报4(3):149~154
4,李英波,罗信昌,李滨等,1995.香菇菌株的限制性片段长度多型性.真菌学报,14(3):209~217
5,周欣起,刘正林,1994.大光木耳栽培技术.江苏食用菌,15(3): 12~14
6,施庆利,罗信昌,1994.RAPD技术在木耳杂交种及原生质体融合子鉴定分析中的应用.中国食用菌,12(6):3~4.
7,秦国夫,贺伟,沈瑞祥,1996.中国密环菌生物种的RAPD分析.真菌学报,15(1):26~33
8,阎培生,罗信昌,周启,1999.木耳属真菌rDNA特异性扩增片段的RFLP研究.菌物系统,18(2): 77~83
9,郭砚翠,刘风春,王雅茹等,1989.大木耳的生物学特性及开发研究.食用菌,5:13~14
10,曾凡亚,张义正,1996.从多糖丰富的样品中制备食用真菌DNA.食用菌学报,3(3): 13~17
11,詹才新,朱兰宝,杨新美,1995.RAPD技术在金针菇杂交育种中的应用.食用菌学报,2(1): 7~12
12,谭琦,1996.利用RAPD技术对不同地区的有害疣孢霉菌株进行遗传分析.食用菌学报,3(4):52~56
13,Bidochka MJ, McDonald MA, Leger RJST, et al., 1994. Differentiation of species and strains of entomopathogenic fungi by random amplifccation of polymorphic DNA. Curr Genet, 25: 107~113