Agrobacterium–mediated Transformation of Kalanchoe laxiflora

作     者：Xiling Wang Xinlu Chen Qunkang Cheng Kaikai Zhu Xiaohan Yang Zongming Cheng Xiling Wang;Xinlu Chen;Qunkang Cheng;Kaikai Zhu;Xiaohan Yang;Zongming Cheng

作者机构：Department of Plant SciencesUniversity of TennesseeKnoxvilleTN37996USA College of BiotechnologySouthwest UniversityChongqing400715China Department of Entomology and Plant PathologyUniversity of TennesseeKnoxvilleTN37996USA College of HorticultureNanjing Agricultural UniversityNanjing210095China Biosciences DivisionOak Ridge National LaboratoryOak RidgeTN37831-6422USA 

出 版 物：《Horticultural Plant Journal》 (园艺学报（英文版）)

年 卷 期：2019年第5卷第5期

页      面：221-228页

核心收录：

学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 090706[农学-园林植物与观赏园艺] 0907[农学-林学] 09[农学] 0807[工学-动力工程及工程热物理] 0901[农学-作物学] 0902[农学-园艺学] 0713[理学-生态学] 

基　　金：supported by the Tennessee Agricultural Experiment Station Hatch Project (TEN00491) Fundamental Research Funds for the Central Universities (XDJK2018C065) 

主　　题：Kalanchoe laxiflora Agrobacterium-mediated transformation CAM plants 

摘      要：Plants with crassulacean acid metabolism(CAM) generally utilize water 20%–80% more efficiently than non-CAM plants. The whole genomes of several CAM plants have been sequenced or are being sequenced. For effective genome characterization and genome editing of CAM plants,an efficient transformation system is essential. In this study, we developed an Agrobacterium-mediated transformation protocol for Kalanchoe laxiflora, an obligate CAM plant,by optimizing several factors affecting the transformation efficiency. Agrobacterium strains AGL1, C58, EHA105,and GV3101 were all suitable for K. laxiflora transformation. Fifty-nine percent of the leaf explants yielded kanamycin-resistant and GUS-positive shoots. Polymerase chain reaction and quantitative real-time PCR(qRT-PCR) using gus A-, gus Plus-, npt II-and hpt-specific primers confirmed that the transgenes were integrated into K. laxiflora genome and expressed. This efficient transformation system will allow effective functional characterization of genes through over-or down-expression, knockout, or genome editing.