|
|
|
| Cloning and Expression Characterization Analysis of the Watermelon (Citrullus lanatus) Fruit Flesh Hardness Regulating Gene ClMADS37 |
| ZHOU Mei-Han1,2, LIU Chen-Xin1,2, LIU Shi-Liang1,2, MA Ting-Ting1,2, JIA Yun-He1,2,3, WANG Xue-Zheng1,2,* |
1 Key Laboratory of Biology and Germplasm Innovation of Horticultural Crops in Northeast China, Ministry of Agriculture and Rural Affairs, Harbin 150030, China; 2 College of Horticulture, Northeast Agricultural University, Harbin 150030, China; 3 Horticulture Branch of Heilongjiang Academy of Agricultural Sciences, Harbin 150030, China |
|
|
|
|
Abstract The MADS-box transcription factor family is widely involved in fruit ripening, cell wall remodeling, and texture regulation in plants, playing a central role in maintaining and softening fruit flesh. However, the molecular mechanisms of this family in regulating flesh hardness in watermelon (Citrullus lanatus) remain unclear. To investigate the regulatory function of MADS-box transcription factors in the formation of fruit flesh firmness in watermelon, fruit flesh tissues of the hard-flesh line '51' and the soft-flesh line '1338' were used as experimental materials. Through transcriptome analysis and qRT-PCR validation, a candidate gene associated with fruit flesh firmness, designated ClMADS37, was screened. The coding sequence of this gene was cloned, and its structural and expression characteristics were analyzed using bioinformatics methods. The results showed that the open reading frame (ORF) of ClMADS37 was 1 176 bp in length, encoding 391 amino acids. The protein was predicted to be an acidic, stable, hydrophilic protein localized in the nucleus. Expression analysis revealed that ClMADS37 expression levels in the flesh were significantly positively correlated with fruit firmness (r=0.7558). Furthermore, its expression was highest in stems and lowest in leaves. Promoter region analysis identified multiple light-responsive and hormone-responsive elements. In summary, ClMADS37 may positively regulate watermelon flesh hardness, and this study provides a theoretical basis and genetic resource for improving fruit texture and molecular breeding in watermelon.
|
|
Received: 29 December 2025
|
|
|
|
Corresponding Authors:
*wangxuezheng@neau.edu.cn
|
|
|
|
[1] 高欢, 郑珂昕, 廖光联, 等. 2023. 中华猕猴桃全基因组MADS-box基因家族鉴定及表达分析[J]. 果树学报, 40(11): 2307-2324. (Gao H, Zheng K X, Liao G L, et al.2023. Genome-wide identification and expression analysis of the MADS-box gene family in Actinidia chinensis[J]. Journal of Fruit Science, 40(11): 2307-2324.) [2] 高磊, 赵胜杰, 路绪强, 等. 2016. 利用SSR标记对西瓜果肉硬度性状的连锁分析[J]. 植物遗传资源学报, (5): 866-870. (Gao L, Zhao S J, Lu X Q, et al. 2016. Linkage analysis of flesh firmness trait in watermelon using SSR markers[J]. Journal of Plant Genetic Resources, (5): 866-870.) [3] 李干琼, 王志丹. 2019. 我国西瓜产业发展现状及趋势分析[J]. 中国瓜菜, 32(12): 79-83. (Li G Q, Wang Z D.2019. Current situation and development trend of watermelon industry in China[J]. China Cucurbits and Vegetables, 32(12): 79-83.) [4] 李钰婷. 2021. 西瓜属4种近缘种果肉质地特征及相关基因表达分析[D]. 硕士毕业论文, 东北农业大学, 导师: 王学征, pp. 11-12. (Li Y T.2021. Analysis of flesh texture character‐istics and related gene expression in four related species of Citrullus[D]. Thesis for M.S., Northeast Agricultural University, Supervisor: Wang X Z, pp. 11-12.) [5] 刘晨昕 .2023. 西瓜ClMADS37基因克隆及在果实发育中的表达分析[D]. 硕士毕业论文, 东北农业大学, 导师: 王学征, pp. 10-12. (Liu C X.2023. Cloning of ClMADS37 gene and its expression analysis during fruit development in watermelon[D]. Thesis for M. S., Northeast Agricultural University, Supervisor: Wang X Z, pp. 10-12.) [6] 刘景安, 何洪巨, 郭绍贵, 等. 2013. 西瓜果实成熟软化的生理生化机制[J]. 果树学报, 30(5): 813-818. (Liu J A, He H J, Guo S G, et al.2013. Physiological and biochemical mechanisms of watermelon fruit ripening and softening[J]. Journal of Fruit Science, 30(5): 813-818.) [7] 柳文, 马永鹏, 应支萍, 等. 2024. 朱红大杜鹃MADS-box基因家族的全基因组鉴定与特征分析[J]. 植物科学学报,42(5): 624-633. (Liu W, Ma Y P, Ying Z P, et al.2024. Genome-wide identification and characteristic analysis of MADS-box gene family in Rhododendron griersonianum[J]. Acta Botanica Sinica, 42(5): 624-633.) [8] 钱金森, 田雅, 王飞鉴, 等. 2022. 基于转录组测序信息的德国鸢尾 MADS-box 基因家族分析[J]. 分子植物育种, 20(1): 49-63. (Qian J S, Tian Y, Wang F J, et al.2022. Analysis of MADS-box gene family in Iris germanica based on transcriptome sequencing data[J]. Journal of Molecular Plant Breeding, 20(1): 49-63.) [9] 申孜, 闫小玲, 郝琴, 等. 2024. 毛竹PheMADS47a基因启动子克隆及功能分析[J]. 核农学报,38(9): 1682-1690. (Shen Z, Yan X L, Hao Q, et al.2024. Cloning and functional analysis of promoter of PheMADS47a gene in Phyllostachys edulis[J]. Journal of Nuclear Agricultural Sciences, 38(9): 1682-1690.) [10] 孙蕾. 2021. 西瓜中心果肉硬度主效QTL及候选基因分析[D]. 博士毕业论文, 东北农业大学, 导师: 栾非时, pp.16-17. (Sun L.2021. Analysis of main effect QTL and candidate genes for firmness of central watermelon flesh[D]. Dissertation for Ph.D., Northeast Agricultural University, Supervisor: Luan F S, pp.16-17.) [11] 王梦园, 魏倩睿, 李海艳, 等. 2025. 辣椒MADS-box转录因子基因CaAGL61的耐热功能分析[J]. 中国农业科学,58(8): 1604-1616. (Wang M Y, Wei Q R, Li H Y, et al.2025. Functional analysis of heat tolerance of MADS-box transcription factor gene CaAGL61 in Capsicum annuum[J]. Scientia Agricultura Sinica, 58(8): 1604-1616.) [12] 张雯, 马依努尔·加马力, 王敏, 等. 2022. 不同葡萄品种果肉质地和细胞结构及生理指标分析[J]. 西北植物学报, 42(11): 1870-1879. (Zhang W, Mayinur J M, Wang M, et al.2022. Analysis of flesh texture, cell structure and physiological indices of different grape varieties[J]. Acta Botanica Boreali-Occidentalia Sinica, 42(11): 1870-1879.) [13] 郑雪梦, 时月, 王雪敏, 等. 2020. 基于转录组测序的西瓜果肉硬度相关基因表达分析[J]. 中国瓜菜, (10): 15-22. (Zheng X M, Shi Y, Wang X M, et al. 2020. Expression analysis of genes related to watermelon flesh firmness based on transcriptome sequencing[J]. China Cucurbits and Vegetables, (10): 15-22.) [14] 朱敬业, 陈建设, 陈勇. 2025. 坚果质构特性与动态感官感知研究进展[J]. 中国食品学报, 25(8): 455-470. (Zhu J Y, Chen J S, Chen Y.2025. Research progress on texture properties and dynamic sensory perception of nuts[J]. Journal of Chinese Institute of Food Science and Technology, 25(8): 455-470.) [15] Arroyo A J M, McQuinn R, Poole M, et al.2009. Fleshy fruit expansion and ripening are regulated by the tomato SHATTERPROOF gene TAGL1[J]. Plant Cell, 21: 3041-3062. [16] Brummell D A, Harpster M H.2001. Cell wall metabolism in fruit softening and quality and its manipulation in transgenic plants[J]. Plant Molecular Biology, 47(1-2): 311-340. [17] Chi Z H, Wang L W, Hu Q K, et al.The MADS-Box transcription factor EjAGL18 negatively regulates malic acid content in loquat (Eriobotrya japonica) by repressing EjtDT1[J]. International Journal of Molecular Sciences, 2025, 26(2): 530. [18] Dong T T, Hu Z L, Deng L, et al.2013. A tomato MADS-box transcription factor, SlMADS1, acts as a negative regulator of fruit ripening[J]. Plant Physiology, 163(2): 1026-1036. [19] Gao Y, Guo Y G, Su Z Y, et al.2020.Transcriptome analysis of genes related to fruit texture in watermelon[J]. Scientia Horticulturae, 262: 109075. [20] Guo L, Luo X, Li M, et al.2022. Mechanism of fertilization-induced auxin synthesis in the endosperm for seed and fruit development[J]. Nature Communications, 13: 3985. [21] Guo P Y, Cheng X, Xing C J, et al.2025. Overexpression of SlMADS48 alters the structure of inflorescence and the sizes of sepal and fruit in tomato[J]. Plants, 14(21): 3259. [22] Guo S G, McQuinn R, Poole M, et al.2015. Comparative transcriptome analysis of cultivated and wild watermelon during fruit development[J]. PLOS ONE, 10(6): e0130267. [23] Guo X H, Li L Z, Han Z P, et al.2026. SlMADS50, a type Ⅰ MADS-box transcription factor, regulates tomato (Solanum lycopersicum) plant architecture via hormonal pathways[J]. Plant Science, 362: 112811. [24] Hu J, Chang X J, Zhang Y, et al.2021. The pineapple MADS-box gene family and the evolution of early monocot flower[J]. Scientific Reports, 11: 849. [25] Qi X L, Liu C L, Song L L, et al.2020. PaMADS7, a MADS-box transcription factor, regulates sweet cherry fruit ripening and softening[J]. Plant Science, 301: 110634 [26] Wang P, Wang S B, Chen Y, et al.2019. Genome-wide analysis of the MADS-box gene family in watermelon[J]. Computational Biology and Chemistry, 80: 341-350. [27] Wang W, Guo S, Tian S, et al.2017. Abscisic acid pathway involved in the regulation of watermelon fruit ripening and quality trait evolution[J]. PLOS ONE, 12: e179944. [28] Wang Z M, Chang J S, Han J, et al.2025.Genome-wide reidentification and expression analysis of MADS-box gene family in cucumber[J]. International Journal of Molecular Sciences, 26(8): 3800. [29] Wu Q F, Wu Y, Li R, et al.2025. Research progress on the regulation of plant floral organ development by the MADS-box gene family[J]. International Journal of Molecular Sciences, 26(18): 8946. [30] Xiao K, Fan J M, Bi X Y, et al.2025. A NAC transcription factor and a MADS-box protein antagonistically regulate sucrose accumulation in strawberry receptacles[J]. Plant Physiology, 197(3)(https://doi.org/10.1093/plphys/kiaf043). [31] Yin W C, Hu Z L, Cui B L, et al.2017. Suppression of the MADS-box gene SlMBP8 accelerates fruit ripening of tomato (Solanum lycopersicum)[J]. Plant Physiology and Biochemistry, 118: 235-244. [32] Yin W C, Yu X H, Chen G P, et al.2018. Suppression of SlMBP15 inhibits plant vegetative growth and delays fruit ripening in tomato[J]. Frontiers in Plant Science, 9: 938. [33] Zheng Y K, Liu M T, Caihong, et al.2020. Characteristics of banana B genome MADS-box family demonstrate their roles in fruit development, ripening, and stress[J]. Scientific Reports, 10: 20840. |
|
|
|