Optimization of protoplast isolation,transformation and its application in sugarcane (Saccharum spontaneum L.)

作     者：Qiongli Wang Guangrun Yu Zhiyong Chen Jinlei Han Yufang Hu Kai Wang Qiongli Wang;Guangrun Yu;Zhiyong Chen;Jinlei Han;Yufang Hu;Kai Wang

作者机构：Key Laboratory of GeneticsBreeding and Multiple Utilization of CropsMinistry of Education/Fujian Provincial Key Laboratory of Haixia Applied Plant Systems BiologyCenter for Genomics and BiotechnologyFujian Agriculture and Forestry UniversityFuzhou 350002FujianChina National Engineering Research Center of SugarcaneFujian Agriculture and Forestry UniversityFuzhou 350002FujianChina 

出 版 物：《The Crop Journal》 (作物学报（英文版）)

年 卷 期：2021年第9卷第1期

页      面：133-142页

核心收录：

学科分类：09[农学] 0901[农学-作物学] 

基　　金：funded by the National Natural Science Foundation of China(3190020451 and 31771862) 

主　　题：Saccharum spontaneum L. Protoplast isolation PEG Transformation efficiency Transient expression 

摘      要：Sugarcane is a prominent source of sugar and ethanol *** analysis for trait improvement of sugarcane is greatly hindered by its complex genome,long breeding cycle,and recalcitrance to genetic *** protoplast-based transient transformation system is a versatile and convenient tool for in vivo functional gene analysis;however,quick and effective transformation systems are still lacking for ***,we developed an efficient protoplast-based transformation system by optimizing conditions of protoplasts isolation and PEG-mediated transformation in *** yield of viable protoplasts was approximately 1.26×107 per gram of leaf material,and the transformation efficiency of 80.19%could be achieved under the optimized ***,using this approach,the nuclear localization of an ABI5-like bZIPs transcription factor was validated,and the promoter activity of several putative DNase I hypersensitive sites(DHSs)was *** results indicated this system can be conveniently applied to protein subcellular localization and promoter activity assays.A highly efficient *** mesophyll cell protoplast isolation and transient transformation method was developed,and it shall be suitable for in vivo functional gene analysis in sugarcane.