Color-related chlorophyll and carotenoid concentrations of Chinese kale can be altered through CRISPR/Cas9 targeted editing of the carotenoid isomerase gene BoaCRTISO

Bo Sun , Min Jiang , Hao Zheng , Yue Jian , Wen-Li Huang , Qiao Yuan , Ai-Hong Zheng , Qing Chen , Yun-Ting Zhang , Yuan-Xiu Lin , Yan Wang , Xiao-Rong Wang , Qiao-Mei Wang , Fen Zhang , Hao-Ru Tang

Horticulture Research ›› 2020, Vol. 7 ›› Issue (1) : 161

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Horticulture Research ›› 2020, Vol. 7 ›› Issue (1) :161 DOI: 10.1038/s41438-020-00379-w
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Color-related chlorophyll and carotenoid concentrations of Chinese kale can be altered through CRISPR/Cas9 targeted editing of the carotenoid isomerase gene BoaCRTISO
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Abstract

The carotenoid isomerase gene (BoaCRTISO) of Chinese kale was targeted and edited using the CRISPR/Cas9 system in the present study. The results showed a high mutation rate (81.25%), and 13 crtiso mutants were obtained. Only two types of mutations, insertions and replacements, were found. Both the total and individual carotenoid and chlorophyll concentrations of the biallelic and homozygous mutants were reduced, and the total levels declined by 11.89–36.33%. The color of the biallelic and homozygous mutants changed from green to yellow, likely reflecting a reduction in the color-masking effect of chlorophyll on carotenoids. The expression levels of most carotenoid and chlorophyll biosynthesis-related genes, including CRTISO, were notably lower in the mutants than in the WT plants. In addition, the functional differences between members of this gene family were discussed. In summary, these findings indicate that CRISPR/Cas9 is a promising technique for the quality improvement of Chinese kale and other Brassica vegetables.

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Bo Sun, Min Jiang, Hao Zheng, Yue Jian, Wen-Li Huang, Qiao Yuan, Ai-Hong Zheng, Qing Chen, Yun-Ting Zhang, Yuan-Xiu Lin, Yan Wang, Xiao-Rong Wang, Qiao-Mei Wang, Fen Zhang, Hao-Ru Tang. Color-related chlorophyll and carotenoid concentrations of Chinese kale can be altered through CRISPR/Cas9 targeted editing of the carotenoid isomerase gene BoaCRTISO. Horticulture Research, 2020, 7 (1) : 161 DOI:10.1038/s41438-020-00379-w

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References

[1]

Sun, B. et al. Molecular cloning and expression analysis of the ζ-carotene desaturase gene in Chinese kale (Brassica oleracea var. alboglabra Bailey). Hortic. Plant J. 4, 94-102 (2018).

[2]

Chen, J. et al. Assessment of glucosinolates in Chinese kale by near-infrared spectroscopy. Int. J. Food Prop. 17, 1668-1679 (2014).

[3]

Sun, B., Yan, H. Z., Liu, N., Wei, J. & Wang, Q. M. Effect of 1-MCP treatment on postharvest quality characters, antioxidants and glucosinolates of Chinese kale. Food Chem. 131, 519-526 (2012).

[4]

Guo, X. Y., Song, C. K., Ho, C. T. & Wan, X. C. Contribution of L-theanine to the formation of 2, 5-dimetheylpyrazine, a key roasted peanutty flavor in Oolong tea during manufacturing processes. Food Chem. 263, 18-28 (2018).

[5]

Li, X. D. et al. Lycopene is enriched in tomato fruit by CRISPR/Cas9-mediated multiplex genome editing. Front. Plant Sci. 9, 559 (2018).

[6]

Ma, J., Li, J. W., Xu, Z. S., Wang, F. & Xiong, A. S. Transcriptome profiling of genes involving in carotenoid biosynthesis and accumulation between leaf and root of carrot (Daucus carota L.). Acta Biochim. Biophys. Sin. 50, 481-490 (2018).

[7]

Lu, S. et al. The cauliflower Or gene encodes a DnaJ cysteine-rich domain-containing protein that mediates high levels of β-carotene accumulation. Plant Cell 18, 3594-3605 (2006).

[8]

Lv, P., Li, N., Liu, H., Gu, H. H. & Zhao, W. E. Changes in carotenoid profiles and in the expression pattern of the genes in carotenoid metabolisms during fruit development and ripening in four watermelon cultivars. Food Chem. 174, 52-59 (2015).

[9]

Kato, M. et al. Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit. Plant Physiol. 134, 824-837 (2004).

[10]

Yamagishi, M., Kishimoto, S. & Nakayama, M. Carotenoid composition and changes in expression of carotenoid biosynthetic genes in tepals of Asiatic hybrid lily. Plant Breed. 129, 100-107 (2010).

[11]

Yuan, H., Zhang, J., Nageswaran, D. & Li, L. Carotenoid metabolism and regulation in horticultural crops. Hortic. Res. 2, 15036 (2015).

[12]

Isaacson, T., Ronen, G., Zamir, D. & Hirschberg, J. Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of β-carotene and xanthophylls in plants. Plant Cell 14, 333-342 (2002).

[13]

Chai, C. L. et al. ZEBRA2, encoding a carotenoid isomerase, is involved in photoprotection in rice. Plant Mol. Biol. 75, 211-221 (2011).

[14]

Su, T. B. et al. Loss of function of the carotenoid isomerase gene BrCRTISO confers orange color to the inner leaves of Chinese cabbage (Brassica rapa L. ssp. pekinensis). Plant Mol. Biol. Rep. 33, 648-659 (2015).

[15]

Jia, H. G. et al. Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker. Plant Biotechnol. J. 15, 817-823 (2016).

[16]

Li, R. et al. CRISPR/Cas9-Mediated SlNPR1 mutagenesis reduces tomato plant drought tolerance. BMC Plant Biol. 19, 38 (2019).

[17]

Braatz, J. et al. CRISPR-Cas9 targeted mutagenesis leads to simultaneous modification of different homoeologous gene copies in polyploid oilseed rape (Brassica napus). Plant Physiol. 174, 935-942 (2017).

[18]

Li, X. F. et al. High-efficiency breeding of early-maturing rice cultivars via CRISPR/Cas9-mediated genome editing. J. Genet. Genom. 44, 175-178 (2017).

[19]

Sun, B. et al. CRISPR/Cas9-mediated mutagenesis of homologous genes in Chinese kale. Sci. Rep. 8, 16786 (2018).

[20]

Sun, B. et al. Functional differences of BaPDS1 and BaPDS2 genes in Chinese kale. R. Soc. Open Sci. 6, 190260 (2019).

[21]

Lei, J. J., Chen, G. J., Chen, C. M. & Cao, B. H. Germplasm diversity of Chinese kale in China. Hortic. Plant J. 3, 101-104 (2017).

[22]

Yan, L. H. et al. High-efficiency genome editing in Arabidopsis using YAO promoter-driven CRISPR/Cas9 system. Mol. Plant 8, 1820-1823 (2015).

[23]

Doyle, J. J. & Doyle, J. L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19, 11-15 (1987).

[24]

Shi, Y. M. et al. Molecular cloning and functional characterization of the lycopene ε-cyclase gene via virus-induced gene silencing and its expression pattern in Nicotiana tabacum. Int. J. Mol. Sci. 15, 14766-14785 (2014).

[25]

Chen, Q. et al. An alternative cetyltrimethylammonium bromide-based protocol for RNA isolation from blackberry (Rubus L.). Genet. Mol. Res. 11, 1773-1782 (2011).

[26]

Hasperué, J. H., Gómez-Lobato, M. E., Chaves, A. R., Civello, P. M. & Martínez, G. A. Time of day at harvest affects the expression of chlorophyll degrading genes during postharvest storage of broccoli. Postharvest Biol. Technol. 82, 22-27 (2013).

[27]

Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method . Methods 25, 402-408 (2001).

[28]

Büchert, A. M., Civello, P. M. & Martínez, G. A. Effect of hot air, UV-C, white light and modified atmosphere treatments on expression of chlorophyll degrading genes in postharvest broccoli (Brassica oleracea L.) florets. Sci. Hortic. 127, 214-219 (2011).

[29]

Tian, S. W. et al. Efficient CRISPR/Cas9-based gene knockout in watermelon. Plant Cell Rep. 36, 399-406 (2017).

[30]

Miao, H. et al. Changes of the photosynthetic pigment and differential expression of the correlated genes in a chlorophyll-deficient cucumber mutant (Cucumis sativus L.). Sci. Agric. Sin. 43, 4027-4035 (2010).

[31]

Gu, H. H. et al. Characterization of the appearance, health-Promoting compounds, and antioxidant capacity of the florets of the loose-curd cauliflower. Int. J. Food Prop. 18, 392-402 (2015).

[32]

Guiné, R. P. F. & Barroca, M. J. Effect of drying treatments on texture and color of vegetables (pumpkin and green pepper). Food Bioprod. Process. 90, 58-63 (2012).

[33]

Lee, S. M., Lee, K. T., Lee, S. H. & Song, J. K. Origin of human colour preference for food. J. Food Eng. 119, 508-515 (2013).

[34]

Manninen, H., Paakki, M., Hopia, A. & Franzén, R. Measuring the green color of vegetables from digital images using image analysis. LWT Food Sci. Technol. 63, 1184-1190 (2015).

[35]

Nisar, N., Li, L., Lu, S., Khin, N. C. & Pogson, B. J. Carotenoid metabolism in plants. Mol. Plant 8, 68-82 (2015).

[36]

Koichi, K. & Tatsuru, M. Transcriptional regulation of tetrapyrrole biosynthesis in Arabidopsis thaliana. Front. Plant Sci. 1811, 1-17 (2016).

[37]

Soufflet-Freslon, V. et al. Functional gene polymorphism to reveal species history: the case of the CRTISO gene in cultivated carrots. PloS ONE 8, e70801 (2013).

[38]

Zhang, B. et al. Disruption of a CAROTENOID CLEAVAGE DIOXYGENASE 4 gene converts flower colour from white to yellow in Brassica species. N. Phytol. 206, 1513-1526 (2015).

[39]

Di, F. F. et al. Genome-wide analysis of the PYL gene family and identification of PYL genes that respond to abiotic stress in Brassica napus. Genes 9, 156 (2018).

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