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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

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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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Acknowledgements

The authors would like to thank lab mates for their kind assistance in the present study.

Funding

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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Correspondence to Zhongliang Su.

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The authors declare no competing interests.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Informed consent was obtained from all individual participants included in the study.

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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

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