古尼蟲(chóng)草小孢變種無(wú)性型原生質(zhì)體制備及再生條件的研究 陳曉琳　 樊美珍　 李增智　 摘　要：以從古尼蟲(chóng)草小孢變種Cordyceps gunnii var.minor上分離的無(wú)性型古尼擬青霉小孢變種Paecilomyces gunnii var.minor G106M為出發(fā)菌株,進(jìn)行原生質(zhì)體制備及再生條件的研究.將培養(yǎng)16h的菌絲體用溶壁酶和蝸牛酶的混合酶液于28℃酶解1.5～2h,原生質(zhì)體產(chǎn)量可達(dá)3.74×107/mL.以培養(yǎng)12h的菌絲制備的原生質(zhì)體在0.6mol/L氯化鈉的SDAY培養(yǎng)基上再生率最高,為17.92%. 關(guān)鍵詞：蟲(chóng)草;蟲(chóng)生真菌;最佳再生條件;再生率 分類號(hào)：Q939.97　文獻(xiàn)標(biāo)識(shí)碼：A 文章編號(hào)：1672-6472(2005)04-0533-0538 PREPARATION AND REGENERATION OF PROTOPLASTS OF PAECILOMYCES GUNNII VAR. MINOR CHEN Xiao-Lin　 FAN Mei-Zhen　 LI Zeng-Zhi　 基金項(xiàng)目：安徽省教育廳自然科學(xué)基金重點(diǎn)項(xiàng)目(編號(hào):2002KJ110ZD) 作者簡(jiǎn)介：樊美珍,通訊作者,E-mail:mzfan@ahau.edu.cn 作者單位：陳曉琳（安徽農(nóng)業(yè)大學(xué),合肥,230036）　 　　　　　樊美珍（安徽農(nóng)業(yè)大學(xué),合肥,230036）　 　　　　　李增智（安徽農(nóng)業(yè)大學(xué),合肥,230036）　 參考文獻(xiàn)： [1]Gao D, 1996. Microbiology Genetics. Shandong: Shandong University Press. 1～377 (in Chinese) [2]Li ZZ, Li CR< Huang B, Fan MZ & Lee MW, 1999. New variety of Cordyceps gunnii(Berk.) Berk. and its Paecilomyces anamorph. The Korea Journal of Mycology, 26 (1): 15～21 [3]Nong XQ, Wu ZK, BAO JZ, 1990. Studies on protoplast preparation and regeneration of Metarhizium anisopliae. Microbiology, 17 (2): 76～80 (in Chinese) [4]Deng CS, Wu ZK, Ye ZC, 1991. Studies on protoplast preparation and regeneration of Verticillium lecanii. Studies and Application of Entomogenous Fungi in China, 2:176～179 (in Chinese) [5]Xiang SH, 1989. Protoplast formation and regeneration of Geotrichum suaveolens. Journal of Microbiology, 9 ( 1 ): 24～30 (in Chinese) [6]Zhang HB, Ding HS, Cheng LZ, Sha T, 1999.Studies on protoplast preparation and regeneration of Aureobasidium pullulans.Journal of Yunnan University, 21 (3): 202～204 (in Chinese) [7]Zhou DP, Ping WX, 1990. Protoplast Fusion of Microorganisms. Heilongjiang: Heilongjiang Technology Press. 1～406 (in Chinese) [8]Riba G, 1978. Combinasion apres heteroceryose chez le champignon entomopathogene Paecilomyces fumosoroseus (Deuteromycete). Entomophaga,23:417～421 [9]Silveira WD, Azevedo JL, 1987. Protoplast fusion and genetic recombination in Metarhuzium anisopliae. Enzyme Microbiol Technol, 9: 149～152 [10]Pfeifer TA, Khachatourians GG, 1987. The formation of protoplasts from Beauveria bassiana. Appl Microbiol Biotechnol, 26:248～253 [11]高東,1996.微生物遺傳學(xué).山東:山東大學(xué)出版社.1～377 [12]農(nóng)向群,吳正鎧,包建中,1990.金龜子綠僵菌原生質(zhì)體形成和再生的研究.微生物學(xué)通報(bào),17(1):76～80 [13]鄧春生,吳正凱,葉正楚,1991.蠟蚧輪枝孢原生質(zhì)體形成和再生的研究.中國(guó)蟲(chóng)生真菌研究與應(yīng)用,2:176～179 [14]向世華,1989.果香地霉原生質(zhì)體的釋放與再生.微生物學(xué)雜志,9(1):24～30 [15]張漢波,丁驊孫,程立忠,沙濤,1999.出芽短梗霉的原生質(zhì)體形成及再生研究.云南大學(xué)學(xué)報(bào)(自然科學(xué)版),21(3):202～204 [16]周東坡,平文祥,1990.微生物原生質(zhì)體融合.黑龍江:黑龍江科學(xué)技術(shù)出版社.1～402 菌物學(xué)報(bào) 2005年第4期-HTM文件 No.10