利用RAPD技術(shù)對木耳屬菌株進(jìn)行分類鑒定的研究
閻培生 羅信昌 周啟
摘 要:利用RAPD技術(shù)對木耳屬不同種和種內(nèi)不同菌株進(jìn)行了分子鑒定。結(jié)果表明,在供試的26個隨機(jī)引物中,有10個引物可擴(kuò)增出清晰、穩(wěn)定的DNA帶型,其中引物S4和S6與供試木耳屬菌株的基因組DNA結(jié)合位點少而保守,擴(kuò)增圖譜具有種的特異性,可用于種的快速準(zhǔn)確鑒定;其綜隨機(jī)引物的擴(kuò)增DNA多態(tài)性較強,可用于種內(nèi)不同菌株的鑒定;聚類分析表明,在75%相似水平下,可將供試的木耳屬菌株分成四大類,其中有三大類各代表一個形態(tài)鑒定的種。研究結(jié)果表明了RAPD技術(shù)可有效地用于木耳屬種或菌株的快速準(zhǔn)確鑒定。
關(guān)鍵詞:隨機(jī)擴(kuò)增多態(tài)性DNA,分子鑒定,木耳屬,聚類分析
中圖分類號:Q936, 文獻(xiàn)標(biāo)識碼:A.
文章編號: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.
國家自然科學(xué)基金資助項目
閻培生(華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室 武漢 430070)(萊陽農(nóng)學(xué)院植保系應(yīng)用真菌研究室 萊陽 265200)
羅信昌(華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室 武漢 430070)
周啟(華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室 武漢 430070)
[參 考 文 獻(xiàn)]
1,劉學(xué)敏,惠東威,張明厚等,1997.大豆灰斑病菌生理小種的RAPD標(biāo)記.菌物系統(tǒng),16(2):128~133
2,張引芳,1994.Studies on the differentiation of Lentinula edodes by RAPD assay. 食用菌學(xué)報,1(1):21~27
3,李麗嘉,1984.木耳屬二新種.真菌學(xué)報4(3):149~154
4,李英波,羅信昌,李濱等,1995.香菇菌株的限制性片段長度多型性.真菌學(xué)報,14(3):209~217
5,周欣起,劉正林,1994.大光木耳栽培技術(shù).江蘇食用菌,15(3): 12~14
6,施慶利,羅信昌,1994.RAPD技術(shù)在木耳雜交種及原生質(zhì)體融合子鑒定分析中的應(yīng)用.中國食用菌,12(6):3~4.
7,秦國夫,賀偉,沈瑞祥,1996.中國密環(huán)菌生物種的RAPD分析.真菌學(xué)報,15(1):26~33
8,閻培生,羅信昌,周啟,1999.木耳屬真菌rDNA特異性擴(kuò)增片段的RFLP研究.菌物系統(tǒng),18(2): 77~83
9,郭硯翠,劉風(fēng)春,王雅茹等,1989.大木耳的生物學(xué)特性及開發(fā)研究.食用菌,5:13~14
10,曾凡亞,張義正,1996.從多糖豐富的樣品中制備食用真菌DNA.食用菌學(xué)報,3(3): 13~17
11,詹才新,朱蘭寶,楊新美,1995.RAPD技術(shù)在金針菇雜交育種中的應(yīng)用.食用菌學(xué)報,2(1): 7~12
12,譚琦,1996.利用RAPD技術(shù)對不同地區(qū)的有害疣孢霉菌株進(jìn)行遺傳分析.食用菌學(xué)報,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