1 Crops Research Institute/Fujian Key Laboratory of Vegetable Genetics and Breeding, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2 Faculty of Forestry, University of British Columbia, Vancouver V6T1Z2, Canada
Abstract：Abscisic acid (ABA) serves as a pivotal hormone in plants, regulating the entire process of plant growth, development, and stress responses. Nine-cis-epoxycarotenoid dioxygenase (NCED) acts as the rate-limiting enzyme in ABA biosynthesis, directly determining its biosynthetic efficiency and content. The identification and analysis of the NCED gene in Hibiscus esculentus are crucial for elucidating the mechanism of ABA-mediated regulation of pod senescence. In this study, the HeNCED gene in H. esculentus was identified and analyzed using bioinformatics. The results revealed that the full length of HeNCED gene (GenBank No. PZ350883) was 2 196 bp, with an open reading frame (ORF) of 1 803 bp encoding 600 amino acids. The protein was localized in the cytoplasm, exhibiting acidic, hydrophilic, and unstable properties. Sequence similarity analysis showed over 88% identity with homologous proteins from Gossypium anomalum, G. raimondii, G. arboreum, and H. syriacus, indicating high conservation. qPCR analysis of pod samples at different developmental stages and post-harvest treated with exogenous ABA and nordihydroguaiaretic acid (NDGA) demonstrated a significant positive correlation between HeNCED expression and ABA content (P<0.01). It was hypothesized that HeNCED regulated secondary wall thickening in H. esculentus pods via modulating ABA synthesis. This study preliminarily revealed the molecular mechanism of HeNCED in Hibiscus esculentus pod aging, which can provide a theoretical basis and genetic resources for molecular breeding, cultivation, and post-harvest storage of H esculentus.
[1] 陈尚武, 张大鹏. 2000. ABA和Fluridone对苹果果实成熟的影响[J]. 植物生理学报, 02: 123-129. (Chen S W, Zhang D P.2000. Effects of ABA and fluridone on apple fruit ripening[J]. Journal of Plant Physiology, 123-129.) [2] 崔萌萌, 孙亮, 王灿磊, 等. 2010. 番茄果实LeNCED2基因的克隆及其表达分析[J]. 中国农业大学学报, 15(6): 20-26. (Cui M M, Sun L, Wang C L, et al.2010. Cloning and expression analysis of LeNCED2 gene in tomato fruit[J]. Journal of China Agricultural University, 15(6): 20-26.) [3] 豆峻岭, 刘文革, 赵胜杰, 等. 2014. 西瓜果实不同发育时期内源激素含量的变化[J]. 中国瓜菜, 27(2): 12-15. (Dou J L, Liu W G, Zhao S J, et al.2014. Changes in endogenous hormone contents in watermelon fruit during different developmental stages[J]. China Cucurbits and Vegetables, 27(2): 12-15.) [4] 高艳, 陈光辉, 陈秀娟, 等. 2014. 植物细胞壁纤维素生物合成的调控[J]. 生物技术通报, 01: 1-7. (Gao Y, Chen G H, Chen X J, et al.2014. Regulation of cellulose biosynthesis in plant cell wall[J]. Biotechnology Bulletin, 01: 1-7.) [5] 龚霄, 姜永超, 周伟, 等. 2018. 黄秋葵研究进展及其应用[J]. 食品工业科技, 39(23): 329-333. (Gong X, Jian Y C, Zhou W, et al.2018. Research progress and application of okra[J]. Science and Technology of Food Industry, 39(23): 329-333.) [6] 郭芳芸. 2024. 银杏木材发育木质素合成基因挖掘及其功能研究[D]. 博士学位论文, 南京林业大学, 导师: 汪贵斌, pp. 18-23. (Guo F Y.2024. Developmental characteristics of wood and analysis of genes related to lignin biosynthesis in Ginkgo biloba L.[D]. Thesis for Ph.D., Nanjing Forestry University, Supervisor: Wang G B, pp. 18-23.) [7] 黄欣. 2017. 异花授粉对'琯溪蜜柚'汁胞内源激素和相关代谢途径基因表达的影响[D]. 硕士学位论文, 福建农林大学, 导师: 王平, pp. 23. (Huang X.2017. Effects of cross pollination on endogenous hormones and gene expression of related metabolic pathways in 'Guanximiyou' (Citrus grandis) juice sacs[D]. Thesis for M.S., Fujian Agriculture and Forestry University, Supervisor: Wang P, pp. 23.) [8] 姜微波, 刘尊英, 冯双庆, 等. 2003. 豌豆苗采后GA、乙烯和ABA处理对纤维合成的调控作用[J]. 园艺学报, 30(03): 296-298. (Jiang W B, Liu Z Y, Feng S Q, et al.Regulations of fiber synthesis in excised pea seedlings by gibberellic acid ethylene and abscisic acid[J]. Acta Horticulturae Sinica, 30(03): 296-298.) [9] 李永平, 陈敏氡, 刘建汀, 等. 2022. 黄秋葵纤维素合酶基因家族鉴定及表达分析[J]. 园艺学报49(01): 73-85. (Li Y P, Chen M D, Liu J T, et al.2022. Identification and expression analysis of CESA gene family in Hibiscus esculentus L[J]. Acta Horticulturae Sinica, 49(01): 73-85.) [10] 罗自生. 2006. 采后竹笋木质化与内源激素的关系[J]. 中国农业科学, 39(04): 792-797. (Luo Z S. 2006. Relationship between postharvest lignification and endogenous hormones in bamboo shoots[J]. Scientia Agricultura Sinica 39(04): 792-797.) [11] 秦一统, 付金元, 肖正璐, 等. 2016. 黄秋葵的营养成分及应用价值[J]. 中国食品工业, 5: 2. (Qin Y T, Fu J Y, Xiao Z L, et al.2016. Nutritional components and application value of Okra[J]. China food industry, 5: 2.) [12] 单承莺, 马世宏, 张卫明. 2012. 保健蔬菜黄秋葵的应用价值与前景[J]. 中国野生植物资源, 31(2): 68-71. (Shan C Y, Ma S H, Zhang W M.2012. Utilization and prospect of okra: A health vegetable[J]. Chinese Wild Plant Resottrces, 31(2): 68-71.) [13] 汤佳, 冯金玲, 杨志坚, 等. 2015. 油茶果实内源激素变化与果实发育的关系[J]. 森林与环境学报, 35(4): 331-336. (Tang J, Feng J L, Yang Z J, et al.2015. Relationship between endogenous hormone changes and fruit development in Camellia oleifera[J]. Journal of Forest and Environment, 35(4): 331-336.) [14] 王春飞, 郁松林, 肖年湘, 等. 2007. 果树果实生长发育细胞学研究进展[J]. 中国农学通报, 7: 386-390. (Wang C F, Yu S L, Xiao N X, et al.2007. Advances in cytological research on fruit growth and development in fruit trees[J]. Chinese Agricultural Science Bulletin, 7: 386-390.) [15] 王向阳. 2005. 椪柑萎缩型枯水病和粒化型枯水病几种生理指标差异的研究[J]. 果树学报, 3: 216-219. (Wang X Y.Study on differences of physiological indexes between atrophic and granulated types of water-deficient disease in Ponkan mandarin[J]. Journal of Fruit Science, 3: 216-219.) [16] 席玙芳, 王向阳, 余挺, 等. 1998. 椪柑枯水后几种生理生化指标的变化[J]. 植物生理学通讯, 5: 345-347. (Xi Y F, Wang X Y, Yu T, et al.1998. Changes of physiological and biochemical indexes after water loss in Ponkan mandarin[J]. Plant Physiology Communications, 5: 345-347.) [17] 谢国芳, 刘娜, 宋易, 等. 2021. 菜豆豆荚发育过程中内源激素与细胞壁代谢的关系[J]. 园艺学报, 48(2): 289-299. (Xie G F, Liu N, Song Y, et al.2021. Relationship between endogenous hormones and cell wall metabolism during pod development of common bean[J]. Acta Horticulturae Sinica, 48(2): 289-299.) [18] 许如意, 罗丰, 袁廷庆, 等. 2011. 不同采摘期对黄秋葵果实性状和品质的影响[J]. 长菜江蔬菜, 2: 18-20. (Xu R Y, Luo F, Yuan T Q, et al.2011. Effect of picking time on the fruit characters of okra[J]. Changjiang Vegetable, 2: 18-20.) [19] 杨洪强, 贾文锁, 张大鹏. 2000. 水分胁迫下湖北海棠根系脂氧合酶活性与ABA积累的关系[J]. 植物学报, 42(03): 244-248. (Yang H Q, Jia W S, Zhang D P.2000. Relationship between lipoxygenase activity and ABA accumulation in roots of Malus hupehensis under water stress[J]. Bulletin of Botanical Research, 42(03): 244-248.) [20] 杨艳敏, 徐长宁, 贾继周. 2001. 棉花(陆地棉)纤维发育的珠心培养体系研究[J]. 作物学报, 27(6): 694-703. (Yang Y M, Xu C N, Jia J Z.2001, A system of ovule subculture for cotton (Gossypium hirsutum L.) fiber development[J]. Acta Agronomica Sinica, 27(6): 694-703.) [21] 杨志芳, 陈常璐, 吕海燕, 等. 2025. ABA调控呼吸跃变型果实成熟机制研究进展[J]. 北方园艺, (05): 123-130. (Yang Z F, Chen C L, Lv H Y, et al. 2025. Research progress on the mechanism of ABA regulating climacteric fruit ripening[J]. Northern Horticulture, (05): 123-130.) [22] 岳翔. 2009. 不结球白菜膳食纤维含量分析及理化特性研究[D]. 硕士学位论文, 南京农业大学, 导师: 朱玉英, 侯喜林, pp. 31. (Yue X.2009. Studies on the analysis of dietary fiber content and physiology and biochemistry characterisics in non-heading chinese cabbage[D]. Thesis for M.S., Nanjing Agricultural University, Supervisor: Zhu Y Y, Hou X L, pp. 31.) [23] 张小英. 2021. 刺梨果实膳食纤维积累过程中相关基因的表达[D]. 硕士学位论文, 贵州大学, 导师: 安华明, pp. 37-40. (Zhang X Y.2021. Expression of related genes during dietary fiber accumulation in Rosa roxburghii fruit[D]. Thesis for M.S., Guizhou University, Supervisor:An H M, pp. 37-40.) [24] 赵阳, 何炎红, 林涛, 等. 2025. 胭脂红杏果实发育过程中内源激素与糖酸代谢的关联研究[J].果树学报, 7: 1-22. (Zhao Y, He Y H, Lin T, et al.2025. Correlation study of endogenous hormones and sugar-acid Metabolism during fruit development in Yanzhihong apricot[J]. Journal of Fruit Science, 7: 1-22.) [25] 赵娟娟. 2013. 柚果实生长过程中内源激素含量变化的研究[D]. 硕士学位论文, 福建农林大学, 导师: 潘东明, pp. 26-32. (Zhao J J.2013. Study on the changes of endogenous hormone contents in Pomelo fruit growth process[D]. Thesis for M.S., Fujian agriculture and Forestry University, Supervisor: Pan D M, pp. 26-32.) [26] 赵秋月. 2020. ABA和ETH对'琯溪蜜柚'粒化汁胞次生壁木质素代谢的影响[D]. 硕士学位论文, 福建农林大学, 导师: 王平, pp. 41. (Zhao Q Y.2020. Study on the effects of secondary wall lignin metabolism in the juice sace of 'Guanxi' Pomelo by ABA and ETH[D]. Thesis for M.S., Fujian Agriculture and Forestry University, Supervisor: Wan P, pp. 41.) [27] 赵玉雪, 朱佳敏, 杨霞, 等. 2021. 核桃青皮中木质素、纤维素、半纤维素测定初报[J]. 贵州林业科技, 49(2): 7-10. (Zhao Y X, Zhu J M, Yang X, et al, 2021. A preliminary report on the determination of lignin, cellulose and hemicellulose in walnut green husk[J]. Guizhou Forestry Science and Technology, 49(2): 7-10.) [28] 曾文静, 杨军, 陈世美杰, 等. 2021. 番茄成熟过程中细胞壁代谢相关基因的表达及外源乙烯和ABA处理对其表达的影响[J]. 农业生物技术学报, 29(06): 1040-1049. (Zeng W J, Yang J, Chen S M J, et al.2021. Expression of cell wall metabolism genes during tomato (Solanum lycopersicum) maturation and the effects of treatment with exogenous C2H4 and ABA on their expression[J]. Journal of Agricultural Biotechnology, 29(06): 1040-1049.) [29] 朱东煌, 黄日升, 蔡顺其, 等. 2015. 植物生长调节剂提高溪蜜柚坐果率的研究与应用综述[J]. 中国热带农业, 4: 110-112. (Zhu D H, Huang R S, Cai S Q, et al.2015. Review on the research and application of plant growth regulators in improving fruit set rate of Ximi pomelo[J]. China Tropical Agriculture, 4: 110-112.) [30] 祝军, 柴璐, 侯柄竹, 等. 2015. 果实中脱落酸的研究进展与展望[J]. 园艺学报, 42(9): 1664-1672. (Zhu J, Chai L, Hou B Z, et al.2015. Advances and perspectives in abscisic acid research in fruits[J]. Acta Horticulturae Sinica, 42(9): 1664-1672.) [31] Chernys J T, Zeevaart J A D.2000. Characterization of the 9-cis-epoxycarotenoid dioxygenase gene family and the regulation of abscisic acid biosynthesis in avocado[J]. Plant Physiology, 124(1): 343-353. [32] Jia H F, Chai Y M, Li C L, et al.2011. Abscisic acid plays an important role in the regulation of strawberry fruit ripening[J]. Plant Physiology, 157(1): 188-199. [33] Lang J, Fu Y, Zhou Y, et al.2021. MYB10-D confers PHS-3D resistance to pre-harvest sprouting by regulating NCED in ABA biosynthesis pathway of wheat[J]. New Phytologist, 230(5): 1940-1952. [34] Luchi, Kobayashi, Yamaguchi-shinozaki, 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. [35] Messing S A J, Gabelli S B, Echeverria I, et al.2010. Structural insights into maize viviparous14, a key enzyme in the biosynthesis of the phytohormone abscisic acid[J]. Plant Cell, 22(9): 2970-2980. [36] Nambara E, Marion-poll A.2005. Abscisic acid biosynthesis and catabolism[J]. Annual Review of Plant Biology, 56(1): 165-185. [37] Sun L, Yuan B, Zhang M, et al.2012. Fruit-specific RNAi-mediated suppression of SlNCED1 increases both lycopene and β-carotene contents in tomato fruit[J]. Journal of Experimental Botany, 63(8): 3097-3108. [38] Tan B C, Joseph L M, Deng Y T, et al.2003. Molecular characterization of the Arabidopsis 9-cis epoxycarotenoid dioxygenase gene family[J]. The Plant Journal, 35(1): 44-56. [39] Vallabhaneni R, Bradbury L M T, Wurtzel E T.2010. The carotenoid dioxygenase gene family in maize, sorghum, and rice[J]. Archives of Biochemistry and Biophysics, 504(1): 104-111.
