Agrobacterium–mediated Transformation of Kalanchoe laxiflora

Xiling Wang, Xinlu Chen, Qunkang Cheng, Kaikai Zhu, Xiaohan Yang, Zongming Cheng

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

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 gusA-, gusPlus-, nptII- 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.

Original languageEnglish
Pages (from-to)221-228
Number of pages8
JournalHorticultural Plant Journal
Volume5
Issue number5
DOIs
StatePublished - Sep 2019

Funding

This research was supported by the Tennessee Agricultural Experiment Station Hatch Project (TEN00491) and Fundamental Research Funds for the Central Universities ( XDJK2018C065 ).We thank Robert Trigiano for critical review of the manuscript.

Keywords

  • Agrobacterium-mediated transformation
  • CAM plants
  • Kalanchoe laxiflora

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