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Cloning and Expression Analysis of a ζ-carotene Desaturase Gene (HeZDS) in Hibiscus esculentus |
LI Yong-Ping, WANG Bin, CHEN Min-Dong, ZHU Hai-Sheng*, WEN Qing-Fang* |
Fujian Key Laboratory of Vegetable Genetics and Breeding / Crops Research Institute, Fujian Academy of Agricultural Sciences / Vegetable Research Center, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China |
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Abstract ζ-carotene desaturase (ZDS) is the rate-limiting enzyme in carotenoid synthesis, catalyzing ζ-carotene to form lycopene. In this study, a ZDS gene was cloned from Hibiscus esculentus by transcriptome sequencing. The full-length cDNA sequence of ZDS was 2 189 bp, which contained a 1 686 bp ORF that encoded 561 amino acids, with a predicted molecular weight of 61.79 kD and a hypothetical isoelectric point (pI) of 6.807. It shared over 90% identity with the homologous proteins from Gossypium arboreum, G. hirsutum, Corchorus capsularis, Durio zibethinus and Theobroma cacao, showing that it was highly conservative. This gene was named HeZDS and the GenBank No. was MG372371. qRT-PCR analysis revealed that HeZDS could be expressed in different tissues of H. esculentus, including roots, stems, leaves, flowers, fruits and pod of Hibiscus esculentus, and the highest expression was found in mature leaves during leaf growth and tender pods (2 d after anthesis) in fruit development, respectively. Moreover, the expression of HeZDS gene had close correlations with carotenoid contents. These results provide a basis for the study of the HeZDS gene function and carotenoid regulation mechanism in H. esculentus.
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Received: 25 October 2019
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Corresponding Authors:
*zhs@163.com;fjvrc@163.com
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[1] 樊荣辉, 黄敏玲, 钟淮钦, 等. 2016. 鹤望兰ζ-胡萝卜素脱氢酶基因SrZDS的克隆及表达分析[J]. 园艺学报, 43(05): 957-965.
(Fan R H, Huang M L, Zhong H Q, et al.2016. Cloning and analysis of ζ-carotene desatura in Strelitzia reginae[J]. Acta Horticulturae Sinica, 43(05): 957-965. )
[2] 季静, 山村三郎, 西原昌宏, 等. 2004. 通过转基因提高β-胡萝卜素生物合成量[J]. 中国生物化学与分子生物学报, (04): 440-444.
(Ji J, Yamamura S, Nishihara M, et al. 2004. Increasement of β-carotene biosynthesis by gene transformation[J]. Chinese Journal of Biochemistry and Molecular Biology, (04): 440-444.)
[3] 李永平, 朱海生, 温庆放, 等. 2010. 草莓ζ-胡萝卜素脱氢酶基因ZDS的克隆及特征分析[J]. 热带亚热带植物学报, 18(06): 670-674.
(Li Y P, Zhu H S, Wen Q F, et al.2010. Cloning and characteristics of ζ-carotene desatura gene in Fragaria ananassa Duchesne[J]. Journal of Trop ical and Sub Tropical Botany, 18(06): 670-674.)
[4] 李永平, 王彬, 陈敏氡, 等. 2019a. 黄秋葵八氢番茄红素脱氢酶基因的克隆与分析[J]. 西北植物学报, 39(07): 1195-1202.
(Li Y P, Wan B, Chen M D, et al.2019. Cloning and expression analysis of a phytoene desaturase gene in Hibiscus esculentus L.[J]. Acta Botanica Boreali-Occidentalia Sinica, 39(07): 1195-1202.)
[5] 李永平, 陈敏氡, 朱海生, 等. 2019b. 黄秋葵八氢番茄红素合成酶PSY基因的克隆与分析[J]. 中国细胞生物学学报, 41(10): 1938-1947.
((Li Y P, Chen M D, Zhu H S, et al.2019. Cloning and Expression Analysis of a Phytoenesynthase Gene in Hibiscus esculentus L[J]. Chinese Journal of Cell Biology, 41(10): 1938-1947.
[6] 梁敏华, 杨震峰, 苏新国, 等. 2018. 桃果实ζ-胡萝卜素脱氢酶基因克隆及表达分析[J]. 南方农业学报, 49(05): 825-831.
(Liang M H, Yang Z F, Su X G, Song C Bet al.2018. Cloning and expression analysis of ζ-carotene desaturasegene in peach fruit[J]. Journal of Southern Agriculture, 49(05): 825-831.)
[7] 刘东祥, 叶花兰, 刘国道. 2006. 14个黄秋葵品系叶黄素和β-胡萝卜素产量比较试验[J]. 中国农学通报, 22(6): 425-427.
(Liu D X, Ye H L, Liu G D.2006. The production of lutein and beta-carotene were tested in 14 Abelmosch esculentu[J]. Chinese Agricultural Science Bulletin, 22(6): 425-427.)
[8] 王柬人, 李锡香, 王海平, 等. 2012. 黄瓜ζ-胡萝卜素脱氢酶基因克隆及表达分析[J]. 分子植物育种, 10(05): 520-527.
(Wang J R, Li X X, Wang H P, et al.2012. Molecular cloning and expression analysis of ζ-carotene desaturase gene from Cucumis sativus L.[J]. Molecular Plant Breeding, 10(05): 520-527.)
[9] Aruna G, Baskaran V.2010. Comparative study on the levels of carotenoids lutein, zeaxanthin and β-carotene in Indian spices of nutritional and medicinal importance[J]. Food Chemistry, 123(2): 404-409.
[10] Breitenbach J, Kuntz M, Takaichi S, et al.1999. Catalytic properties of an expressed and purified higher plant type δ-carotene desaturase from Capsicum annuum[J]. European Journal of Biochemistry, 265: 376-383.
[11] Chan-Leon A C, Estrella Maldonado H, Dube P, et al.2017. The high content of betacarotene present in orange-pulp fruits of Carica papaya L. is not correlated with a high expression of the CpLCY-beta2 gene[J]. Food Research International, 100(2): 45-56.
[12] Clotault J, Peltier D, Berruyer R, et al.2008. Expression of carotenoid biosynthesis genes during carrot root development[J]. Journal of Experimental Botany, 59: 3563-3573.
[13] Ji J, Wang G, Wang J H, et al.2009. Function analysis of multiple carotenogenic genes from Lycium barbarum and Gentiana lutea L. for their effects onβ-carotene production in transgenic tobacco[J]. Biotechnology Letters, 31(2): 305-312.
[14] Kim Y S, Park S J, Cho Y H, Park J, et al.2011. Efflect of combined treatment of high hydrostatic pressure and mild heat on the quality of carrot juice[J]. Journal of Food Science, 66(9): 1355-1360.
[15] Li R, Kang C, Song X, et al.2017. A zeta-carotene desaturase gene, IbZDS, in-creases beta-carotene and lutein contents and enhances salt tolerance in transgenic sweetpotato[J]. Plant Science, 262: 39-51.
[16] Mann V, Harker M, Pecker I, et al.2000. Metabolic engineering of astaxanthin production in tobacco flowers[J]. Nature Biotechnology, 18(8): 88-92.
[17] Mayo-Wilson E, Imdad A, Herzer K, et al.2011. Vitamin A supplements for preventing mortality, illness and blindness in children aged under 5: Systematic review and meta-analysis[J]. British Medical Journal, 343(8): 5094-5112.
[18] Mueller L, Boehm V.2011. Antioxidant activity of β-carotene compounds in different in vitro assays[J]. Molecules, 16(2): 55-69.
[19] Oliver J, Palou A.2000. Chromatographic determination of carotenoids in foods[J]. Journal of Chromatography A, 881(1/2): 543-555.
[20] Potter A R.1997. Reducing vitamin a deficiency: Could save the eyesight and lives of countless children[J]. British Medical Journal, 314(7077): 317-318.
[21] Rodrigo M J, Marcos J F, Alferez F, et al.2003. Characterization of Pinalate, a novel Citrus sinensis mutant with afruit-specific alteration that results in yellow pigmentation and decreased ABA content[J]. Experimental Botany, 54(383): 727-738.
[22] Semba R D.2007. Vitamin A and immunity to viral, bacterial and protozoan infections[J]. Proceedings of the Nutrition Society,?58(3): 719-727.
[23] Steinbrenner J, Linden H.2001. Regulation of two carotenoid biosynthesis genes coding for phytoene synthase and carotenoid hydroxylase during stress-induced astaxanthin formation in the green alga Haematococcus pluvialis[J]. Plant Physiology, 125(2): 810-817.
[24] Tang G W, Hu Y M, Yin S A, et al.2012. β-Carotene in Golden Rice is as good as β-carotene in oil at providing vitamin A to children[J]. American Journal of Clinical Nutrition, 96(3): 658-664.
[25] Wisutiamonkul A, Ampomah-Dwamena D, Allan A C,et al.2017. Carotenoid accumulation in durian (Durio zibethinus) fruit is affected by ethylene via modulation of carotenoid pathway gene expression[J]. Plant Physiology & Biochemistry, 115(3):308-319.
[26] Yan P, Gao X Z, Shen W T, et al.2011. Cloning and expression analysis of phytoene desaturase and δ-carotene desaturase genes in Carica papaya[J]. Molecular Biology reports, 38(2): 785-791.
[27] Yan P, Gao X Z, Shen W T, et al.2010. Cloning and expression analysis of phytoene desaturase and ζ-carotene desaturase genes in Carica papaya[J]. Molecular Biology Reports, 38(2): 785-791.
[28] Zanfini A, Corbini G, La Rosa C, et al.2010. Antioxidant activity of tomato lipophilic extracts and interactions between carotenoids and a-tocopherol in synthetic mixtures[J]. LWT-Food Science and Technology, 43: 67-72. |
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