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Engineering chloroplast development in rice through cell-specific control of endogenous genetic circuits

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Published
  • Dong-Yeon Lee
  • Lei Hua
  • Roxana Khoshravesh
  • Rita Giuliani
  • Indrajit Kumar
  • Asaph Cousins
  • Tammy L Sage
  • Julian M Hibberd
  • Thomas P Brutnell
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<mark>Journal publication date</mark>30/11/2021
<mark>Journal</mark>Plant Biotechnology Journal
Issue number11
Volume19
Number of pages13
Pages (from-to)2291-2303
Publication StatusPublished
Early online date18/08/21
<mark>Original language</mark>English

Abstract

The engineering of C4 photosynthetic activity into the C3 plant rice has the potential to nearly double rice yields. To engineer a two-cell photosynthetic system in rice, the rice bundle sheath (BS) must be rewired to enhance photosynthetic capacity. Here, we show that BS chloroplast biogenesis is enhanced when the transcriptional activator, Oryza sativa Cytokinin GATA transcription factor 1 (OsCGA1), is driven by a vascular specific promoter. Ectopic expression of OsCGA1 resulted in increased BS chloroplast planar area and increased expression of photosynthesis-associated nuclear genes (PhANG), required for the biogenesis of photosynthetically active chloroplasts in BS cells of rice. A further refinement using a DNAse dead Cas9 (dCas9) activation module driven by the same cell-type specific promoter, directed enhanced chloroplast development of the BS cells when gRNA sequences were delivered by the dCas9 module to the promoter of the endogenous OsCGA1 gene. Single gRNA expression was sufficient to mediate the transactivation of both the endogenous gene and a transgenic GUS reporter fused with OsCGA1 promoter. Our results illustrate the potential for tissue-specific dCas9-activation and the co-regulation of genes needed for multistep engineering of C4 rice.

Bibliographic note

© 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.