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Title:
Enhanced translation of a chloroplast-expressed RbcS gene restores small subunit levels and photosynthesis in nuclear RbcS antisense plants
Authors:
Dhingra, Amit; Portis, Archie R., Jr.; Daniell, Henry
Affiliation:
AA(Department of Molecular Biology and Microbiology, University of Central Florida, 4000 Central Florida Boulevard, Biomolecular Science, Building 20, Room 336, Orlando, FL 32816-2364; and Photosynthesis Research Unit, Agricultural Research Service, United States Department of Agriculture, and Departments of Crop Sciences and Plant Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801), AB(Department of Molecular Biology and Microbiology, University of Central Florida, 4000 Central Florida Boulevard, Biomolecular Science, Building 20, Room 336, Orlando, FL 32816-2364; and Photosynthesis Research Unit, Agricultural Research Service, United States Department of Agriculture, and Departments of Crop Sciences and Plant Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801), AC(Department of Molecular Biology and Microbiology, University of Central Florida, 4000 Central Florida Boulevard, Biomolecular Science, Building 20, Room 336, Orlando, FL 32816-2364; and Photosynthesis Research Unit, Agricultural Research Service, United States Department of Agriculture, and Departments of Crop Sciences and Plant Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801)
Publication:
Proceedings of the National Academy of Sciences of the United States of America, Volume 101, Issue 16, 2004, pp.6315-6320
Publication Date:
04/2004
Category:
Plant Biology
Origin:
PNAS
DOI:
10.1073/pnas.0400981101
Bibliographic Code:
2004PNAS..101.6315D

Abstract

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a key enzyme that converts atmospheric carbon to food and supports life on this planet. Its low catalytic activity and specificity for oxygen leads to photorespiration, severely limiting photosynthesis and crop productivity. Consequently, Rubisco is a primary target for genetic engineering. Separate localization of the genes in the nuclear and chloroplast genomes and a complex assembly process resulting in a very low catalytic activity of hybrid Rubisco enzymes have rendered several earlier attempts of Rubisco engineering unsuccessful. Here we demonstrate that the RbcS gene, when integrated at a transcriptionally active spacer region of the chloroplast genome, in a nuclear RbcS antisense line and expressed under the regulation of heterologous (gene 10) or native (psbA) UTRs, results in the assembly of a functional holoenzyme and normal plant growth under ambient CO2 conditions, fully shortcircuiting nuclear control of gene regulation. There was ≈150-fold more RbcS transcript in chloroplast transgenic lines when compared with the nuclear RbcS antisense line, whereas the wild type has 7-fold more transcript. The small subunit protein levels in the gene 10/RbcS and psbA/RbcS plants were 60% and 106%, respectively, of the wild type. Photosynthesis of gene 10/RbcS plants was approximately double that of the antisense plants, whereas that of psbA/RbcS plants was restored almost completely to the wild-type rates. These results have opened an avenue for using chloroplast engineering for the evaluation of foreign Rubisco genes in planta that eventually can result in achieving efficient photosynthesis and increased crop productivity.
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