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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 seek to realize
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 to create biallelic indel mutagenesis to reveal
previously unknown roles of blue light receptor phototropin 2 (KfePHOT2)
in the CAM pathway. Knocking-out of the KfePHOT2 reduced stomatal
conductance and CO2 fixation in the late afternoon and increased stomatal
conductance and CO2 fixation in the early evening, indicating that blue
light signaling plays an important role in the CAM pathway. Lastly, we
provide a genome-wide gRNA database targeting 45,183 protein-coding genes
annotated in the K. fedtschenkoi genome.
207 views reported since publication in 2020.