Preparation and Regeneration Conditions of Ceratocystis paradoxa Protoplast  

作　　者：Niu Xiaoqing Yu Fengyu Wang Yenan Tang Qinghua Zhu Hui Qin Weiquan 

机构地区：[1]Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wenchang 571339,China

出　　处：《Plant Diseases and Pests》2019年第5期7-10,共4页植物病虫害研究（英文版）

基　　金：Supported by Key Project of Hainan Province(ZDYF2019072);Basic Scientific Research Project of CATAS(16301520190010).

摘　　要：[Objective]The paper was to reveal the preparation and regeneration conditions of Ceratocystis caradoxa protoplast.[Method]Using multi-factors orthogonal test,the effects of hypha age,enzyme system,time of hydrolysis,hydrolysis temperature,osmotic pressure stabilizer and regeneration medium on C.paradoxa protoplast were studied.[Result]The optimum condition for preparing protoplast were conidia cultured in liguid SYM medium for 24 h,enzyme mixture of 1%driselase and 1%lytic enzyme used to digest hypha for 1.5 h,0.7 mol/L MgSO4·7H2O used as osmotic stabilizer,or individual enzyme 1%driselase used to digest hypha for 1.5 h,1.0 mol/L mannitol as osmotic stabilizer.The regeneration rate was over 40%under C.paradoxa protoplast regenerating on CM medium with mannitol and hydrolysis for 2 h.[Conclusion]Higher protoplast yield and higher regeneration rate could be obtained under above conditions,which is beneficial for transformation and further research.[Objective] The paper was to reveal the preparation and regeneration conditions of Ceratocystis caradoxa protoplast. [Method] Using multi-factors orthogonal test, the effects of hypha age, enzyme system, time of hydrolysis, hydrolysis temperature, osmotic pressure stabilizer and re-generation medium on C. paradoxa protoplast were studied. [Result] The optimum condition for preparing protoplast were conidia cultured in liguid SYM medium for 24 h, enzyme mixture of 1% driselase and 1% lytic enzyme used to digest hypha for 1.5 h, 0.7 mol/L MgSO4·7H2O used as osmotic stabilizer, or individual enzyme 1% driselase used to digest hypha for 1.5 h, 1.0 mol/L mannitol as osmotic stabilizer. The regeneration rate was over 40% under C. paradoxa protoplast regenerating on CM medium with mannitol and hydrolysis for 2 h. [Conclusion] Higher protoplast yield and higher regeneration rate could be obtained under above conditions, which is beneficial for transformation and further research.

关 键 词：CERATOCYSTIS paradoxa PROTOPLAST REGENERATION 

分 类 号：S432.44[农业科学—植物病理学]