1 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2 College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China
Abstract：Molybdenum cofactor sulfurase (MCSU or LOS5) of plants plays an important role in resisting non-biological stress resistance. At present, few studies reported on the regulation of abscisic acid (ABA) synthesis by VvMCSU gene in response to light stress in grape (Vitis vinfera). VvMCSU gene was cloned by PCR with a full length of coding region, then predicted the gene function from the encoded amino acid sequence by bioinformatics. Subsequently, the expression profiles of VvMCSU in response to light shading stress in leaves were analyzed by qPCR in combination with their leaf photosynthetic characteristics. The results were shown below: Under heavy shading, the maximum photosynthetic quantum yield (Fv/Fm) and non-photochemical quenching parameter (qN) increased by 4.32 % and 28.13 % respectively, and the leaf width and the content of ABA significantly increased by 10.16% and by 1.50 times compared with the control group. The coding region of VvMCSU gene was 2 478 bp, which encodes 825 amino acids, and its promoter region had several cis elements in response to abiotic stress and hormone. VvMCSU was a hydrophilic protein with MCSU structure, which interacted with proteins related to ABA synthesis, and belongs to the same evolutionary branch as camellia (Camelia sinensis). The expression of VvMCSU was up-regulated under 60 % light shading, which was 2.77 times higher than that of the control group 0 % shading or under natural light; and 4.01 times higher than that of 80% severe shading. The expression of VvMCSU gene was positively correlated with ABA content and ratios Fv/Fm (P<0.05). In this study, chlorophyll fluorescence and ABA content were used to analyze the expression of VvMCSU under different light conditions. The results suggested that VvMCSU was an important regulatory gene involved in ABA metabolism in response to light shading stress of grape. This study provides a theoretical basis for viniculture studies to improve grape shading tolerance.
[1] 董贞芬, 门宇恒, 邹秋滢, 等. 2019. 叶绿素荧光成像在植物生长环境胁迫中的应用[J]. 沈阳农业大学学报, 50(02): 250-256. (Dong Z F, Men Y H, Zou Q Y, et al.2019. Application of chlorophyll fluorescence imaging in plant growth environment stress[J]. Journal of Shenyang Agricultural University, 50(02): 250-256.) [2] 高学焕, 付凤玲, 牟巍, 等. 2014. 玉米钼辅助因子硫酸化酶基因启动子的克隆与功能验证[J]. 遗传, 36(06): 584-591. (Gao X H, Fu F L, Mu W, et al.2014. Cloning and functional validation of promoter of mo-molybdopterin cofactor sulfurase gene in maize[J]. Hereditas, 36(06): 584-591.) [3] 古丽加汗•克热木, 古丽孜叶•哈力克, 肯吉古丽•苏力旦, 等. 2016. 脱落酸对设施栽培火焰无核葡萄果实品质的影响[J].中外葡萄与葡萄酒,2016(02): 15-18. (Guljahan K, Gulziya H, Kanjigul S, et al.Effect of exogenous ABA on the fruit quality of Flame Seedless grapesvine under greenhouse cultivation[J]. Sino-Overseas Grapevine & Wine, 2016(02): 15-18.) [4] 何静雯, 赵晟, 岳庆春, 等. 2018. 弱光胁迫下'鄞红'葡萄光合特性及相关基因的表达[J]. 西南农业学报, 31(12): 2520-2526. (He J W, Zhao S, Yue Q C, et al.2018. Effects of weak light stress on photosynthetic characteristics and relative gene expression of 'Yinhong' grape[J]. Southwest China Journal of Agricultural Sciences, 31(12): 2520-2526.) [5] 毛剑飞, 李凯伟, 杨再强, 等. 2018. 弱光胁迫及强光恢复对设施红地球葡萄叶片光合及衰老特性的影响[J].江苏农业科学, 46(06): 105-111. (Mao J F, Li K W, Yan Z Q, et al.2018. Effects of weak light stress and strong light recovery on photosynthetic and senescence characteristics of grape leaves in greenhouse[J]. Jiangsu Agricultural Sciences, 46(06): 105-111.) [6] (Ningbo Ecological Environment Meteorological Center. Ningbo climate bulletin for August 2020[R/OL]. Ningbo. 2020-09-04/2022-06-13.) http://www.qx121.com/weather/weatherStatic/climateReport/692.html [7] 丘立杭, 李强, 黄杏, 等. 2017. 弱光胁迫影响甘蔗叶片内源激素的平衡和分蘖进程[J]. 植物生理学报, 53(02): 280-290. (Qiu L H, Li Q, Huang X, et al.2017. Weak-light stress caused by shading affects the balance of endogenous hormones in leaves and tillering process in sugarcane (Saccharum officinarum)[J]. Plant Physiology Journal, 53(02): 280-290.) [8] 魏晓玲, 柯玉琴, 李文卿, 等. 2021. 不同光温条件对烟株叶片活性氧代谢的影响研究[J]. 江西农业学报, 33(08): 59-64. (Wei X L, Ke Y Q, Li W Q, et al.2021. Effects of different light and temperature conditions on reactive oxygen metabolism in tobacco leaves[J]. Acta Agriculturae Jiangxi, 33(08): 59-64.) [9] 周娜娜, 冯素萍, 高新生, 等. 2019. 植物光合作用的光抑制研究进展[J]. 中国农学通报, 35(15): 116-123. (Zhou N N, Feng S P, Gao X S, et al.2019. Photoinhibition of plants photosynthesis: research progress[J]. Chinese Agricultural Science Bulletin, 35(15): 116-123) [10] 朱延姝, 冯辉. 2005. 弱光胁迫下番茄苗期功能叶片内源激素含量的变化[J]. 东北师大学报(自然科学版), 37(03): 77-80. (Zhu Y S, Feng H.2005. Change of internal phyohromons contents in function leaves of tomato seeding under low light stress[J]. Journal of Northeast Normal University (Natural Science Edition), 37(03): 77-80.) [11] Bian Z, Wang Y, Zhang X, et al.2021. A transcriptome analysis revealing the new insight of greenlight on tomato plant growth and drought stress tolerance[J]. Frontiers in Plant Science, 12: 649283. [12] Bittner F, Oreb M, Mendel R R.2001. ABA3 is a molybdenum cofactor sulfurase required for activation of aldehyde oxidase and xanthine dehydrogenase in Arabidopsis thaliana[J]. The Journal of Biological Chemistry, 276(44): 40381-403814. [13] De Wit M, Galvão V C, Fankhauser C.2016. Light-mediated hormonal regulation of plant growth and development[J]. Annual Review of Plant Biology, 67: 513-537. [14] Devireddy A R, Zandalinas S I, Gómez-Cadenas A, et al.2018. Coordinating the overall stomatal response of plants: Rapid leaf-to-leaf communication during light stress[J]. Science Signaling, 11(518): eaam9514. [15] Yadukrishnan P, Datta S.2021. Light and abscisic acid interplay in early seedling development[J]. The New Phytologist, 229(2): 763-769. [16] Guan C F, Ji J, Zhang X Q, et al.2015. Positive feedback regulation of a Lycium chinense-derived VDE gene by drought-induced endogenous ABA, and over-expression of this VDE gene improve drought-induced photo-damage in Arabidopsis[J]. Journal of Plant Physiology, 175: 26-36. [17] Huang J, Zhao X, Chory J.2019. The Arabidopsis transcriptome responds specifically and dynamically to high light stress[J]. Cell Reports, 29(12): 4186-4199. [18] Iuchi S, Kobayashi M, Yamaguchi-Shinozaki K, et al.2000. A stress-inducible gene for 9-cis-epoxycarotenoid dioxygenase involved in abscisic acid biosynthesis under water stress in drought-tolerant cowpea[J]. Plant Physiology, 123(2): 553-562. [19] Jia H, Lu C.2003. Effects of abscisic acid on photoinhibition in maize plants[J]. Plant Science, 165(6): 1403-1410. [20] Lee S U, Mun B G, Bae E K, et al.2021. Drought stress-mediated transcriptome profile reveals NCED as a key player modulating drought tolerance in Populus davidiana[J]. Frontiers in Plant Science, 12: 755539. [21] Lehrke M, Rump S, Heidenreich T, et al.2012. Identification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis[J]. The Biochemical Journal, 441(3): 823-832. [22] Li Q, Yu X, Chen L, et al.2021. Genome-wide identification and expression analysis of the NCED family in cotton (Gossypium hirsutum L.)[J]. Public Library of Science One, 16(2): e0246021. [23] Li Y, Zhang J, Zhang J, et al.2013. Expression of an Arabidopsis molybdenum cofactor sulphurase gene in soybean enhances drought tolerance and increases yield under field conditions[J]. Plant Biotechnology Journal, 11(6): 747-758. [24] Lu Y, Li Y, Zhang J, et al.2013. Overexpression of Arabidopsis molybdenum cofactor sulfurase gene confers drought tolerance in maize (Zea mays L.)[J]. Public Library of Science One, 8(1): e52126. [25] Pei X, Wang X, Fu G, et al.2021. Identification and functional analysis of 9-cis-epoxy carotenoid dioxygenase (NCED) homologs in G. hirsutum[J]. International Journal of Biological Macromolecules, 182: 298-310. [26] Schwarz G, Mendel R R.2006. Molybdenum cofactor biosynthesis and molybdenum enzymes[J]. The Annual Review of Plant Biology, 57: 623-647. [27] Schwartz S H, Léon-Kloosterziel K M, Koornneef M, et al.1997. Biochemical characterization of the aba2 and aba3 mutants in Arabidopsis thaliana[J]. Plant Physiology, 114(1): 161-166. [28] Seo M, Koiwai H, Akaba S, et al.2000. Abscisic aldehyde oxidase in leaves of Arabidopsis thaliana[J]. The Plant Journal, 23(4): 481-488. [29] Suzuki N, Devireddy A R, Inupakutika M A, et al.2015. Ultra-fast alterations in mRNA levels uncover multiple players in light stress acclimation in plants[J]. The Plant Journal, 84(4): 760-772. [30] Xiong L, Ishitani M, Lee H, et al.2001. The Arabidopsis LOS5/ABA3 locus encodes a molybdenum cofactor sulfurase and modulates cold stress- and osmotic stress-responsive gene expression[J]. Plant Cell, 13(9): 2063-2083. [31] Xu D, Li J, Gangappa S N, et al.2014. Convergence of light and ABA signaling on the ABI5 promoter[J]. PLoS Genetics, 10(2): e1004197. [32] Xu D, Wu D, Li X H, et al.2020. Light andabscisic acid coordinately regulate greening of seedlings[J]. Plant Physiology, 183(3): 1281-1294. [33] Yu H Q, Zhang Y Y, Yong T M, et al.2015. Cloning and functional validation of molybdenum cofactor sulfurase gene from Ammopiptanthus nanus[J]. Plant Cell Reports, 34(7): 1165-1176. [34] Yue Y, Zhang M, Zhang J, et al.2011. Arabidopsis LOS5/ABA3 overexpression in transgenic tobacco (Nicotiana tabacum cv. Xanthi-nc) results in enhanced drought tolerance[J]. Plant Science, 181(4): 405-411. [35] Yue Y, Zhang M, Zhang J, et al.2012. Overexpression of the AtLOS5 gene increased abscisic acid level and drought tolerance in transgenic cotton[J]. Journal of Experimental Botany, 63(10): 3741-3748. [36] Zhang J, Yu H, Zhang Y, et al.2016. Increased abscisic acid levels in transgenic maize overexpressing AtLOS5 mediated root ion fluxes and leaf water status under salt stress[J]. Journal of Experimental Botany, 67(5): 1339-1355.
