CRISPR/Cas9-mediated targeted mutagenesis for functional genomics research of crassulacean acid metabolism plants. (28th September 2019)

• Record Type: Journal Article
• Authors: Liu, Degao; Chen, Mei; Mendoza, Brian; Cheng, Hua; Hu, Rongbin; Li, Linling; Trinh, Cong T; Tuskan, Gerald A; Yang, Xiaohan
• Editors: Hultine, Kevin
• Is Part Of: Journal of experimental botany. Volume 70:Number 22(2019)
• Page Start: 6621
• Page End: 6629
• Publication Date: 2019-09-28
• Subjects: Crassulacean acid metabolism -- CRISPR/Cas9 -- functional genomics -- genome editing -- phototropin 2; Botany -- Periodicals; Botany, Experimental -- Periodicals; Plant physiology -- Periodicals; 580
• DOI: 10.1093/jxb/erz415
• Languages: English
• ISSNs: 0022-0957

Abstract

Abstract: Crassulacean acid metabolism (CAM) is an important photosynthetic pathway in diverse lineages of plants featuring high water-use efficiency and drought tolerance. A big challenge facing the CAM research community is to understand the function of the annotated genes in CAM plant genomes. Recently, a new genome editing technology using CRISPR/Cas9 has become a more precise and powerful tool than traditional approaches for functional genomics research in C3 and C4 plants. In this study, we explore the potential of CRISPR/Cas9 to characterize the function of CAM-related genes in the model CAM species Kalanchoë fedtschenkoi . We demonstrate that CRISPR/Cas9 is effective in creating biallelic indel mutagenesis to reveal previously unknown roles of blue light receptor phototropin 2 ( KfePHOT2 ) in the CAM pathway. Knocking out KfePHOT2 reduced stomatal conductance and CO2 fixation in late afternoon and increased stomatal conductance and CO2 fixation during the night, indicating that blue light signaling plays an important role in the CAM pathway. Lastly, we provide a genome-wide guide RNA database targeting 45 183 protein-coding transcripts annotated in the K. fedtschenkoi genome. Abstract : Loss-of-function analysis using CRISPR/Cas9-mediated mutagenesis provides new insight into the role of phototropin 2 in crassulacean acid metabolism