Transcriptomic and physiological analysis of OsCAO1 knockout lines using the CRISPR/Cas9 system in rice
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ORIGINAL ARTICLE
Transcriptomic and physiological analysis of OsCAO1 knockout lines using the CRISPR/Cas9 system in rice Yu Jin Jung1,2 · Hyo Ju Lee1 · Jihyeon Yu3 · Sangsu Bae3 · Yong‑Gu Cho4 · Kwon Kyoo Kang1,2 Received: 29 June 2020 / Accepted: 17 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Key message The altered rice leaf color based on the knockout of CAO1 gene generated using CRISPR/Cas9 technology plays important roles in chlorophyll degradation and ROS scavenging to regulate both natural and induced senescence in rice. Abstract Rice chlorophyllide a oxygenase (OsCAO1), identified as the chlorophyll b synthesis under light condition, plays a critical role in regulating rice plant photosynthesis. In this study, the development of edited lines with pale green leaves by knockout of OsCAO1 gene known as a chlorophyll synthesis process is reported. Eighty-one genetically edited lines out of 181 T0 plants were generated through CRISPR/Cas9 system. The edited lines have short narrow flag leaves and pale green leaves compared with wild-type ‘Dongjin’ plants (WT). Additionally, edited lines have lower chlorophyll b and carotenoid contents both at seedling and mature stages. A transcriptome analysis identified 580 up-regulated and 206 downregulated genes in the edited lines. The differentially expressed genes (DEGs) involved in chlorophyll biosynthesis, magnesium chelatase subunit (CHLH), and glutamate-1-semialdehyde2, 1-aminomutase (GSA) metabolism decreased significantly. Meanwhile, the gel consistency (GC) levels of rice grains, chalkiness ratios and chalkiness degrees (CD) decreased in the edited lines. Thus, knockout of OsCAO1 influenced growth period, leaf development and grain quality characters of rice. Overall, the result suggests that OsCAO1 also plays important roles in chlorophyll degradation and ROS scavenging to regulate both natural and induced rice senescence. Keywords Chlorophyllide a oxygenase (CAO) · CRISPR/Cas9 · Chlorophyll degradation · Transcriptome analysis Abbreviations LHC Chlorophyll–protein complex CAO Chlorophyllide a oxygenase KO Knockout GSA Glutamate-1-semialdehyde2, 1-aminomutase
CHLH Magnesium-chelatase subunit GC Gel consistency CD Chalkiness degrees T-DNA Transfer, DNA
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00299-020-02607-y) contains supplementary material, which is available to authorized users. * Kwon Kyoo Kang [email protected] 1
Division of Horticultural Biotechnology, Hankyong National University, Anseong 17579, South Korea
2
Institute of Genetic Engineering, Hankyong National University, Anseong 17579, South Korea
3
Department of Chemistry, Hanyang University, Seoul 04763, South Korea
4
Department of Crop Science, Chungbuk National University, Cheongju 28644, South Korea
Chloroplasts include chlorophyll and many other proteinbinding pigments, which are responsible for capturing light energy needed for photosynthesis. Green chloro
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