Agrobacterium–mediated Transformation of Kalanchoe laxiflora

Journal Article · Wed Jul 10 00:00:00 EDT 2019 · Horticultural Plant Journal

DOI:https://doi.org/10.1016/j.hpj.2019.07.001· OSTI ID:1561638

Wang, Xiling ^[1]; Chen, Xinlu ^[2]; Cheng, Qunkang ^[2]; Zhu, Kaikai ^[3]; Yang, Xiaohan ^[4]; Cheng, Zongming ^[2]

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1. Univ. of Tennessee, Knoxville, TN (United States); Southwest Univ., Chongqing (China)
2. Univ. of Tennessee, Knoxville, TN (United States)
3. Nanjing Univ. (China)
4. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

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.

• Research Organization: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
• Sponsoring Organization: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
• Grant/Contract Number: AC05-00OR22725
• OSTI ID: 1561638
• Journal Information: Horticultural Plant Journal, Journal Name: Horticultural Plant Journal Journal Issue: 5 Vol. 5; ISSN 2468-0141
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CAM plants
Kalanchoe laxiflor aAgrobacterium-mediated transformation