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Modifying plant photosynthesis and growth via simultaneous chloroplast transformation of rubisco large and small subunits

Lookup NU author(s): Dr Maxim KapralovORCiD

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Abstract

© 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.

Publication metadata

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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