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Osmanthus fragrans OfLCYB Increased Carotenoid Content and Resistance to Low Temperature Stress in Transgenic Tobacco (Nicotiana tabacum) |
PENG Lin*, WANG Yi-Guang*, DONG Bin, ZHONG Shi-Wei, FANG Qiu, XIAO Zheng, DENG Jin-Ping, ZHAO Hong-Bo** |
School of Landscape Architecture/Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants/Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants, Zhejiang A & F University, Hangzhou 311300, China |
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Abstract Lycopene β-cyclase (LCYB) is a key enzyme that catalyzed lycopene in carotenoid biosynthesis pathway. Osmanthus fragrans is mainly distributed in the south of China. O. fragrans, which bloom in late autumn and winter will show a phenomenon of deepening color when faced with low temperatures. In this study, it was found that the petal color of O. fragrans became darker, the carotenoid content increased significantly, and the expression of key synthetic gene OfLCYB was up-regulated by relatively low temperature treatment. To study the mechanism of OfLCYB accumulation and response to carotenoids, in this study, OfLCYB gene (GenBank No. PP438593) was cloned from O. fragrans petals, and overexpression vector was constructed to transform tobacco (Nicotiana tabacum). The results showed that, overexpression of OfLCYB could promote the accumulation of carotenoids in the leaves of transgenic tobacco, and promote the expression of related genes involved in carotenoid biosynthesis pathway. After low temperature treatment (4 ℃), tobacco leaves with OfLCYB overexpression showed better low temperature tolerance compared with control plants.Under low temperature treatment, relative electrolyte leakage (REL) and malondialdehyde (MDA) contents in leaves of transgenic plants were significantly lower than those of control plants. The activity of Superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and proline content were significantly increased. In addition, compared with control plants, the expression levels of 2 cold response transcription factors NtDREB1 (dehydration responsive element binding 1) and NtDREB3, and 2 cold induction genes NtERD10A (early responsive to dehydration 10A) and NtERD10B in transgenic tobacco leaves were up-regulated. According to the results, it was speculated that OfLCYB might enhance plant low temperature tolerance by promoting carotenoid synthesis. This study provides an reference for improving cold tolerance of O. fragrans and expanding its cultivation area.
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Received: 26 December 2023
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Corresponding Authors:
** zhaohb@zafu.edu.cn
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About author:: * These authors contributed equally to this work |
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[1] 高俊凤. 2006. 植物生理学实验指导[M]. 北京: 高等教育出版社, pp. 211-228. (Gao J F.2006. Experimental Guidance of Plant Physiology[M]. Higher Education Press, Beijing, China, pp. 211-228.) [2] 何静娟, 范燕萍. 2022. 观赏植物花色相关的类胡萝卜素组成及代谢调控研究进展[J]. 园艺学报, 48(12): 1-10. (He J J, Fan Y P.2022. Progress in composition and metabolic regulation of carotenoids related to floral color[J]. Acta Horticulturae Sinica, 48(12): 1-10. [3] 卢素文. 2019. 柑橘番茄红素β-环化酶基因功能分析及其转录调控研究[D]. 博士学位论文, 华中农业大学, 导师: 邓秀新, pp. 9-10. (Lu S W.2019. Functional identification and transcriptional regulatory analysis of citrus lycopene β-cyclase genes[D]. Thesis for Ph. D., Huazhong Agricultural University, Supervisor: Deng X X, pp. 9-10.) [4] 向其柏, 刘玉莲. 2008. 中国桂花品种图志[M]. 杭州: 浙江科学技术出版社, pp. 86-88. (Xiang Q B, Liu Y L.2008. An Illustrated Monograph of the Sweet Osmanthus Variety in China[M]. Zhejiang Science & Technology Press, Hangzhou, China, pp. 86-88.) [5] 杨康民. 2013. 中国桂花[M]. 北京: 中国林业出版社, pp. 005-007. (Yang K M.2013. Chinese Osmanthus[M]. China Forestry Publishing House, Beijing, China, pp. 005-007.) [6] Ackerveken G V D.2017. Seeing is believing: Imaging the delivery of pathogen effectors during plant infection[J]. New Phytologist, 216(1): 205-215. [7] Aguilera J, Randez-Gil F, Prieto, J A.2007. Cold response in Saccharomyces cerevisiae: New functions for old mechanisms[J]. FEMS Microbiology Reviews, 31: 327-341. [8] Ahmad A, Hadi F, Ali N.2014. Effective phytoextraction of cadmium (Cd) with increasing concentration of total phenolics and free proline in Cannabis sativa (L) plant under various treatments of fertilizers, plant growth regulators and sodium salt[J]. International Journal of Phytoremediation, 17(1): 56-65. [9] Bhattacharjee S.2005. Reactive oxygen species and oxidative burst: Roles in stress, senescence and signal transducation in plants[J]. Review Articles, 89: 7. [10] Breton G, Danyluk J, Charron J B F, et al.2003. Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis[J]. Plant Physiology, 132(1): 64-74. [11] Catalá R, Santos E, Alonso J M, et al.2003. Mutations in the Ca2+/H+ transporter CAX1 increase CBF/DREB1 expression and the cold-acclimation response in Arabidopsis[J]. The Plant Cell, 15(12): 2940-2951. [12] Cazzonelli C I, Cuttriss A J, Cossetto S B, et al.2009. Regulation of carotenoid composition and shoot branching in Arabidopsis by a chromatin modifying histone methyltransferase, SDG8[J]. The Plant Cell, 21(1): 39-53. [13] Chen L, Zhong H, Ren F, et al.2011a. A novel cold-regulated gene, COR25, of Brassica napus is involved in plant response and tolerance to cold stress[J]. Plant Cell Reports, 30(4): 463-471. [14] Chen X Y, Han H P, Jiang P, et al.2011b. Transformation of β-Lycopene cyclase genes from Salicornia europaea and Arabidopsis conferred salt tolerance in Arabidopsis and tobacco[J]. Plant & Cell Physiology, 52(5): 909-921. [15] Chinnusamy V, Ohta M, Kanrar S, et al.2003. ICE1: A regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis[J]. Genes & Development, 17(8): 1043-1054. [16] Cuttriss A J, Chubb A C, Alawady A, et al.2007. Regulation of lutein biosynthesis and prolamellar body formation in Arabidopsis[J]. Functional Plant Biology, 34(8): 663-672. [17] Gechev T S, Dinakar C, Benina M, et al.2012. Molecular mechanisms of desiccation tolerance in resurrection plants[J]. Cellular and Molecular Life Sciences, 69(19): 3175-3186. [18] Gonnet J F.2001. Colour effects of co-pigmentation of anthocyanin revisited-3. A further description using CIELAB differences and assessment of matched colours using the CMC model[J]. Food Chemistry, 75(4): 473-485. [19] Guo X Y, Zhang L, Dong G Q, et al.2019. A novel cold-regulated protein isolated from Saussurea involucrata confers cold and drought tolerance in transgenic tobacco (Nicotiana tabacum)[J]. Plant Science, 289: 110246. [20] Hasanuzzaman M, Nahar K, Alam M, et al.2013. Physiological, biochemical, and molecular mechanisms of heat stress tolerance in plants[J]. International Journal of Molecular Sciences, 14(5): 9643-9684. [21] Heidarvand L, Maali A R.2010. What happens in plant molecular responses to cold stress?[J]. Acta Physiologiae Plantarum, 32(3): 419-431. [22] Jaspers P, Kangasjärvi J.2010. Reactive oxygen species in abiotic stress signaling[J]. Physiologia Plantarum, 138: 405-413. [23] Kang C, Zhai H, Xue L Y, et al.2018. A lycopene β-cyclase gene, IbLCYB2, enhances carotenoid contents and abiotic stress tolerance in transgenic sweetpotato[J]. Plant Science, 272: 243-254. [24] Kasuga M, Miura S, Shinozaki K, et al.2004. A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought- and low-temperature stress tolerance in tobacco by gene transfer[J]. Plant & Cell Physiology, 45(3): 346-350. [25] Khan M Z, Takemura M, Maoka T, et al.2016. Carotenoid analysis of sweetpotato Ipomoea batatas and functional identification of its lycopene β- and ε-cyclase genes[J]. Zeitschrift für Naturforschung C, 71(9-10): 313-322. [26] Knight M R, Knight H.2012. Low-temperature perception leading to gene expression and cold tolerance in higher plants[J]. New Phytologist, 195(4): 737-751. [27] Kössler S, Armarego-Marriott T, Tarkowská D, et al.2021. Lycopene β-cyclase expression influences plant physiology, development, and metabolism in tobacco plants[J]. Journal of Experimental Botany, 72(7): 2533-2569. [28] Llorente F, López-Cobollo R M, Catalá R, et al.2002. A novel cold-inducible gene from Arabidopsis, RCI3, encodes a peroxidase that constitutes a component for stress tolerance[J]. The Plant Journal, 32(1): 13-24. [29] Li R J, Kang C, Song X J, et al.2017. A ζ-carotene desaturase gene, IbZDS, increases β-carotene and lutein contents and enhances salt tolerance in transgenic sweetpotato[J]. Plant Science, 262: 39-51. [30] Moreno J C, Cerda A, Simpson K, et al.2016. Increased Nicotiana tabacum fitness through positive regulation of carotenoid, gibberellin and chlorophyll pathways promoted by Daucus carota lycopene β-cyclase (Dclcyb1)expression[J]. Journal of Experimental Botany, 67(8): 2325-2338. [31] Nakayama K, Okawa K, Kakizaki T, et al.2007. Arabidopsis Cor15am is a chloroplast stromal protein that has cryoprotective activity and forms oligomers[J]. Plant Physiology, 144(1): 513-523. [32] Ning G, Xiao X, Lv H, et al.2012. Shortening tobacco life cycle accelerates functional gene identification in genomic research[J]. Plant Biology, 14(6): 934-943. [33] Pulido P, Toledo-Ortiz G, Phillips M A, et al.2013. Arabidopsis J-protein J20 delivers the first enzyme of the plastidial isoprenoid pathway to protein quality control[J]. The Plant Cell, 25(10): 4183-4194. [34] Qing H S, Chen J H, Jiang L L, et al.2022. Functional characterization of two lycopene cyclases from sweet Osmanthus (Osmanthus fragrans)[J]. Scientia Horticulturae, 299: 111062. [35] Ralley L, Schuch W, Fraser P D, et al.2016. Genetic modification of tomato with the tobacco lycopene β-cyclase gene produces high β-carotene and lycopene fruit[J]. Zeitschrift für Naturforschung C, 71(9-10): 295-301. [36] Shi Y M, Guo J G, Zhang W, et al.2015. Cloning of the lycopene β-cyclase gene in Nicotiana tabacum and its overexpression confers salt and drought tolerance[J]. International Journal of Molecular Sciences, 16(12): 30438-30457. [37] Song X Y, Zhu W J, Tang R M, et al.2016. Over-expression of StLCYb increases β-carotene accumulation in potato tubers[J]. J Plant Biotechnology Reports, 10(2): 95-104. [38] Steponkus P L, Uemura M, Joseph R A, et al.1998. Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana[J]. Proceedings of the National Academy of Sciences of the USA, 95(24): 14570-14575. [39] Sun T H, Yuan H, Cao H B, et al.2017. Carotenoid metabolism in plants: The role of plastids[J]. Molecular Plant, 11(1): 58-74. [40] Thalhammer A, Bryant G, Sulpice R, et al.2014. Disordered cold regulated15 proteins protect chloroplast membranes during freezing through binding and folding, but do not stabilize chloroplast enzymes in vivo[J]. Plant Physiology, 166(1): 190-201. [41] Thalhammer A, Hincha D K.2014. A mechanistic model of COR15 protein function in plant freezing tolerance: Integration of structural and functional characteristics[J]. Plant Signaling & Behavior, 9(12): e977722. [42] Thomashow M F.1998. Role of cold-responsive genes in plant freezing tolerance1[J]. Plant Physiology, 118(1): 1-8. [43] Thomashow M F.2010. Molecular basis of plant cold acclimation: Insights gained from studying the CBF cold response pathway[J]. Plant Physiology, 154(2): 571-577. [44] Wan F X, Pan Y, Li J H, et al.2014. Heterologous expression of Arabidopsis C-repeat binding factor 3 (AtCBF3) and cold-regulated 15A (AtCOR15A) enhanced chilling tolerance in transgenic eggplant (Solanum melongena L.)[J]. 33(12): 1951-1961. [45] Wang K, Zhong M, Wu Y H, et al.2017. Overexpression of a chrysanthemum transcription factor gene DgNAC1 improves the salinity tolerance in chrysanthemum[J]. Plant Cell Reports, 36: 571-581. [46] Wang M Q, Huang Q X, Lin P, et al.2020. The overexpression of a transcription factor gene VbWRKY32 enhances the cold tolerance in Verbena bonariensis[J]. Frontiers in Plant Science, 10: 1746. [47] Wang Y G, Dong B, Wang N N, et al.2022a. A WRKY transcription factor PmWRKY57 from Prunus mume improves cold tolerance in Arabidopsis thaliana[J]. Molecular Biotechnology, 65(8): 1359-1368. [48] Wang Y Q, Yang Y, Fei Z J, et al.2013. Proteomic analysis of chromoplasts from six crop species reveals insights into chromoplast function and development[J]. Journal of Experimental Botany. 64(4): 949-961. [49] Wang Y G, Zhang C, Dong B, et al.2018. Carotenoid accumulation and its contribution to flower coloration of Osmanthus fragrans[J]. Frontiers in Plant Science, 9: 1499. [50] Wang Y G, Zhang C, Xu B, et al.2022b. Temperature regulation of carotenoid accumulation in the petals of sweet Osmanthus via modulating expression of carotenoid biosynthesis and degradation genes[J]. BMC Genomics, 23(1): 418. [51] Xiong L M, Zhu J K.2003. Regulation of abscisic acid biosynthesis[J]. Plant Physiology, 133(1): 29-36. [52] Yuan H M, Sheng Y, Chen W J, et al.2017. Overexpression of Hevea brasiliensis HbiCE1 enhances cold tolerance in Arabidopsis[J]. Frontiers in Plant Science, 8: 1462. [53] Zarka D G, Vogel J T, Cook D, et al.2003. Cold induction of Arabidopsis CBF genes involves multiple ICE (inducer of CBF expression) promoter elements and a cold-regulatory circuit that is desensitized by low temperature[J]. Plant Physiology, 133(2): 910-918. [54] Zeng J, Wang C, Chen X, et al.2015. The lycopene β-cyclase plays a significant role in provitamin A biosynthesis in wheat endosperm[J]. BMC Plant Biology, 15(1): 112. [55] Zhang L H, Sun L, Zhang L, et al.2017. A Cu/Zn superoxide dismutase gene from Saussurea involucrata Kar. & Kir., SiCSD, enhances drought, cold, and oxidative stress in transgenic tobacco[J]. 97(5): 816-826. [56] Zhao L Y, Yang T Y, Xing C H, et al.2019. The β-amylase PbrBAM3 from pear (Pyrus betulaefolia) regulates soluble sugar accumulation and ROS homeostasis in response to cold stress[J]. Plant Science, 287: 110184. [57] Zhou M Q, Wu L H, Liang J, et al.2012. Cold-induced modulation of CbICE53 gene activates endogenous genes to enhance acclimation in transgenic tobacco[J]. Molecular Breeding, 30(4): 1611-1620. [58] Zhou Q Q, Li Q C, Li P, et al.2019. Carotenoid cleavage dioxygenases: Identification, expression, and evolutionary analysis of this gene family in tobacco[J]. International Journal of Molecular Sciences, 20(22): 5796. [59] Zhu J K.2002. Salt and drought stress signal transduction in plants[J]. Annual Review of Plant Biology, 53(1): 247-273. |
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