Abstract
Carotenoids are important photosynthetic pigments with many physiological functions, nutritional properties and high commercial value. β-carotene hydroxylase is one of the key enzymes in the carotenoid synthesis pathway of Chlamydomonas reinhardtii for the conversion of β-carotene to astaxanthin. The vector p64DZ containing the β-carotene hydroxylase gene crtZ from Haematococcus pluvialis was transformed into C. reinhardtii CC-503. The transformants were selected by alternate culture in solid–liquid medium containing spectinomycin (100 µg mL−1). PCR results indicated that the gene crtZ and aadA were integrated into the genome of C. reinhardtii. RT-PCR analysis showed that the gene crtZ was transcribed in Chlamydomonas transformants. HPLC analysis showed that the content of astaxanthin and β-carotene in cells of C. reinhardtii were simultaneously increased. Under medium light intensity cultivation (60 µmol m−2 s−1), transgenic C. reinhardtii had an 85.8% increase in β-carotene content compared with the wild type. The content of astaxanthin and β-carotene reached 1.97 ± 0.13 mg g−1 fresh cell weight (FCW) and 105.94 ± 5.84 µg g−1 FCW, which were increased 18% and 42.4% than the wild type after 6 h of high light treatment (200 µmol m−2 s−1), respectively. Our results indicate the regulatory effect on pigments in C. reinhardtii by β-carotene hydroxylase gene of H. pluvialis, and demonstrate the positive effect of high light stress on pigment accumulation in transgenic C. reinhardtii.
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The authors would like to thank lab mates for their kind assistance in the present study.
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This work was supported by the National Natural Science Foundation of China (Grant No. 41471279).
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ZS conceived and designed research. KH conducted experiments and wrote the manuscript. MH, YW and MW analyzed data. All authors read and approved the manuscript.
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Huang, K., Su, Z., He, M. et al. Simultaneous accumulation of astaxanthin and β-carotene in Chlamydomonas reinhardtii by the introduction of foreign β-carotene hydroxylase gene in response to high light stress. Biotechnol Lett 44, 321–331 (2022). https://doi.org/10.1007/s10529-022-03230-5
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DOI: https://doi.org/10.1007/s10529-022-03230-5