Journal
PLANT JOURNAL
Volume 85, Issue 1, Pages 148-160Publisher
WILEY-BLACKWELL
DOI: 10.1111/tpj.13098
Keywords
Rubisco; photosynthesis; CO2 concentration mechanism; chloroplast transformation; Synechococcus elongatus; Nicotiana tabacum
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Funding
- UK Biotechnology and Biological Sciences Research Council [BB/I024488/1]
- US National Science Foundation [EF-1105584]
- National Institute of General Medical Sciences of the National Institutes of Health (USA) [F32GM103019]
- 20:20 Wheat Institute Strategic Program (Biotechnology and Biological Sciences Research Council) [BB/J/00426X/1]
- Cornell University Biotechnology Resource Center (National Institutes of Health) [S10RR025502]
- BBSRC [BB/I024488/1, BB/N013662/1, BBS/E/C/00005202, BB/I017372/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BBS/E/C/00005202, BB/I024488/1, BB/N013662/1, BB/I017372/1] Funding Source: researchfish
- Natural Environment Research Council [ceh010010] Funding Source: researchfish
- NATIONAL CENTER FOR RESEARCH RESOURCES [S10RR025502] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [F32GM103019] Funding Source: NIH RePORTER
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Introducing a carbon-concentrating mechanism and a faster Rubisco enzyme from cyanobacteria into higher plant chloroplasts may improve photosynthetic performance by increasing the rate of CO2 fixation while decreasing losses caused by photorespiration. We previously demonstrated that tobacco plants grow photoautotrophically using Rubisco from Synechococcus elongatus, although the plants exhibited considerably slower growth than wild-type and required supplementary CO2. Because of concerns that vascular plant assembly factors may not be adequate for assembly of a cyanobacterial Rubisco, prior transgenic plants included the cyanobacterial chaperone RbcX or the carboxysomal protein CcmM35. Here we show that neither RbcX nor CcmM35 is needed for assembly of active cyanobacterial Rubisco. Furthermore, by altering the gene regulatory sequences on the Rubisco transgenes, cyanobacterial Rubisco expression was enhanced and the transgenic plants grew at near wild-type growth rates, although still requiring elevated CO2. We performed detailed kinetic characterization of the enzymes produced with and without the RbcX and CcmM35 cyanobacterial proteins. These transgenic plants exhibit photosynthetic characteristics that confirm the predicted benefits of introduction of non-native forms of Rubisco with higher carboxylation rate constants in vascular plants and the potential nitrogen-use efficiency that may be achieved provided that adequate CO2 is available near the enzyme.
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