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Lookup NU author(s): Dr Maxim KapralovORCiD
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© 2020 ASPB.Engineering improved Rubisco for the enhancement of photosynthesis is challenged by the alternate locations of the chloroplast rbcL gene and nuclear RbcS genes. Here we develop an RNAi-RbcS tobacco (Nicotiana tabacum) master-line, tobRrDS, for producing homogenous plant Rubisco by rbcL-rbcS operon chloroplast transformation. Four genotypes encoding alternative rbcS genes and adjoining 59-intergenic sequences revealed that Rubisco production was highest (50% of the wild type) in the lines incorporating a rbcS gene whose codon use and 59 untranslated-region matched rbcL. Additional tobacco genotypes produced here incorporated differing potato (Solanum tuberosum) rbcL-rbcS operons that either encoded one of three mesophyll small subunits (pS1, pS2, and pS3) or the potato trichome pST-subunit. The pS3-subunit caused impairment of potato Rubisco production by ;15% relative to the lines producing pS1, pS2, or pST. However, the bAbB loop Asn-55-His and Lys-57-Ser substitutions in the pS3-subunit improved carboxylation rates by 13% and carboxylation efficiency (CE) by 17%, relative to potato Rubisco incorporating pS1 or pS2-subunits. Tobacco photosynthesis and growth were most impaired in lines producing potato Rubisco incorporating the pST-subunit, which reduced CE and CO2/O2 specificity 40% and 15%, respectively. Returning the rbcS gene to the plant plastome provides an effective bioengineering chassis for introduction and evaluation of novel homogeneous Rubisco complexes in a whole plant context.
Author(s): Martin-Avila E, Lim Y-L, Birch R, Dirk LMA, Buck S, Rhodes T, Sharwood RE, Kapralov MV, Whitney SM
Publication type: Article
Publication status: Published
Journal: Plant Cell
Year: 2020
Volume: 32
Issue: 9
Pages: 2898-2916
Online publication date: 04/09/2020
Acceptance date: 02/04/2016
ISSN (print): 1040-4651
ISSN (electronic): 1532-298X
Publisher: American Society of Plant Biologists
URL: https://doi.org/10.1105/tpc.20.00288
DOI: 10.1105/TPC.20.00288
PubMed id: 32647068
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