Pham, H.A.; Cho, K.; Tran, A.D.; Chandra, D.; So, J.; Nguyen, H.T.T.; Sang, H.; Lee, J.-Y.; Han, O. Compensatory Modulation of Seed Storage Protein Synthesis and Alteration of Starch Accumulation by Selective Editing of 13 kDa Prolamin Genes by CRISPR-Cas9 in Rice. Int. J. Mol. Sci.2024, 25, 6579.
Pham, H.A.; Cho, K.; Tran, A.D.; Chandra, D.; So, J.; Nguyen, H.T.T.; Sang, H.; Lee, J.-Y.; Han, O. Compensatory Modulation of Seed Storage Protein Synthesis and Alteration of Starch Accumulation by Selective Editing of 13 kDa Prolamin Genes by CRISPR-Cas9 in Rice. Int. J. Mol. Sci. 2024, 25, 6579.
Pham, H.A.; Cho, K.; Tran, A.D.; Chandra, D.; So, J.; Nguyen, H.T.T.; Sang, H.; Lee, J.-Y.; Han, O. Compensatory Modulation of Seed Storage Protein Synthesis and Alteration of Starch Accumulation by Selective Editing of 13 kDa Prolamin Genes by CRISPR-Cas9 in Rice. Int. J. Mol. Sci.2024, 25, 6579.
Pham, H.A.; Cho, K.; Tran, A.D.; Chandra, D.; So, J.; Nguyen, H.T.T.; Sang, H.; Lee, J.-Y.; Han, O. Compensatory Modulation of Seed Storage Protein Synthesis and Alteration of Starch Accumulation by Selective Editing of 13 kDa Prolamin Genes by CRISPR-Cas9 in Rice. Int. J. Mol. Sci. 2024, 25, 6579.
Abstract
Rice prolamins are categorized into three groups by molecular size (10, 13, or 16 kDa), while the 13 kDa prolamins are assigned to four subgroups (Pro13a-I, Pro13a-II, Pro13b-I, and Pro13b-II) based on cysteine residue content. Here, we generated four knockout strains of rice using CRISPR-Cas9, each of which demonstrated selectively reduced expression of specific subgroups of the 13 kDa prolamins. These four mutant rice lines also demonstrated compensatory expression of glutelins and non-targeted prolamins, which resulted in low grain weight, altered starch content and atypically-shaped starch granules and protein bodies. Transcriptome analysis identified 746 transcripts that were differentially expressed in the mutant rice lines during development. These transcripts played roles primarily in RNA processing, protein synthesis, stress response, and transport. After selective suppression/knockout of genes in the Pro13a-I subgroup, compensatory upregulation of genes in the Pro13a-II and Pro13b-I/II subgroups was observed. Compensatory expression of 9 ER stress and 17 transcription factor genes was also observed in mutant rice in which expression of 13 kDa prolamin genes was suppressed. Our results provide valuable insight into the regulatory mechanisms underlying rice seed development and the specific roles played by 13 kDa rice prolamins and their target genes.
Keywords
prolamin; 13 kDa prolamin; glutelin; seed storage protein; CRISPR-Cas9; protein body; starch; rice seed; ER stress; differentially expressed genes
Subject
Biology and Life Sciences, Plant Sciences
Copyright:
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