赵竟男;江梅;苏晓庆;
ZHAO Jingnan,JIANG Mei,SU Xiaoqing(Department of Biology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

• 1:贵阳医学院生物学教研室
• 2:贵阳医学院生物学教研室
• 3:贵阳医学院生物学教研室 贵州贵阳550004
• 4:贵州贵阳550004

摘要(Abstract)：

目的:建立贵阳腐霉原生质体的制备和再生系统,为该菌的基因工程和细胞工程改良提供适宜的真菌状态。方法:对影响原生质体形成的各因素,包括菌龄、消化酶的种类和比例、消化时间以及渗透压稳定剂的成分和浓度进行实验比较。结果:以1%纤维素酶与1%溶壁酶1:1组合联合脱壁,0.6mol/L的甘露醇作渗透压稳定剂,酶解采用KPYG2培养基培养52～54h的贵阳腐霉菌菌丝,消化5h所获得的原生质体数最多,原生质体浓度可达到6.48×106个/ml;在0.6mol/L甘露醇作渗透压稳定剂的KPYG2培养基上,原生质体再生率为0.043%。结论:本实验所总结的原生质体制备方法可以获得在基因转化或细胞突变诱导所需的原生质体浓度。
Objective:To provide a suitable fungal material for protoplast-mediated genetic transformation and mutation induction of Pythium guiyangense Su.Methods:Conditions for the fungal protoplast preparation and regeneration including mycelium incubation time,various compounding of enzymes and osmotic stabilizers were examined.Results:Under the conditions as following:mixture of 1% lywallzyme and 1%cellulose(1:1) as digestive enzyme solution;0.6mol/L D-Mannitol as osmotic stabilizer,and 5h,of digestive time the obtained protoplast amount reached 6.48×106 each milliliter.A regeneration rate of 0.043% was obtained.Conclusion:A satisfied concentration of P.guiyangense protoplasts can be achieved with the preparation system developed in this research.

关键词(KeyWords)： 真菌;贵阳腐霉;原生质体;生物灭蚊;渗透压
fungi;Pythium guiyangense Su;protoplasts;mosquito biocontrol;osmotic pressure

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30760140);; 贵州省科技攻关项目〔黔科合NY字(2006)3040号〕

作者(Author): 赵竟男;江梅;苏晓庆;
ZHAO Jingnan,JIANG Mei,SU Xiaoqing(Department of Biology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

Email:

DOI: 10.19367/j.cnki.1000-2707.2008.02.001

参考文献(References)：

• [1]Su Xiao-qing.Anewspecies of Pythium isolated from mos-quito larvae and its ITS region of rDNA[J].Mycosystema,2006(4):523-528.
• [2]Emerson S,Emerson M R.Production,reproduction and reversion of protoplastlike structure in the osmotic strain of Neurospora crassa[J].Proe Nal Acad Sel USA,1958(44):668-671.
• [3]甘志被,赵学慧.黑曲霉ANS-11原生质体的制备[J].湖北农业科学,1994(4):32-34.
• [4]张志光.真菌原生质体技术[M].长沙:湖南科学技术出版社,2003:2,41,70.
• [5]姚婷婷,王正祥.黑曲霉原生质体的制备、再生及转化条件[J].食品与生物技术学报,2006(4):116-120.
• [6]王伟霞,李福后.茯苓原生质体制备与再生条件研究[J].菌物研究,2006(4):65-68.
• [7]何社辉,苏晓庆.灭蚊真菌-大链壶菌原生质体形成和再生的研究[J].真菌学报,1995(3):196-201.

文章评论(Comment)：