不同倍性西瓜发育过程中瓜氨酸合成途径主要氨基酸含量变化及相关基因表达分析

Changes of main amino acid contents in citrulline biosynthesis pathway and expression analysis of related genes during fruit development of watermelon with different ploidy

摘要

以同源二倍体、三倍体和四倍体西瓜伊选为试验材料，研究3种倍性西瓜果实发育过程中瓜氨酸合成途径中谷氨酸、鸟氨酸、瓜氨酸和精氨酸4种氨基酸的含量差异以及其合成途径中关键酶基因的表达差异。结果表明，多倍体西瓜果实中瓜氨酸与鸟氨酸含量均显著高于二倍体，而多倍体精氨酸含量低于二倍体。在西瓜果实发育32 d时，瓜氨酸与鸟氨酸含量表现为三倍体＞四倍体＞二倍体，而谷氨酸与精氨酸含量表现为二倍体＞四倍体＞三倍体。多倍体与二倍体在果实发育后期(24~32 d) NAGS、 NAGK、 GAT、 OTC基因表达量差异显著，二倍体中 ASS1、 ASS3、 ARG基因表达量在整个果实发育期始终高于多倍体。以上结果表明，不同倍性西瓜果实中瓜氨酸含量存在差异可能是瓜氨酸合成途径上下游基因的差异表达造成的。研究结果对揭示不同倍性西瓜中瓜氨酸含量差异机制具有重要意义。

Abstract

The content of four amino acids, glutamic acid, ornithine, citrulline and arginine, as well as the expression differences of key enzyme genes in the citrulline synthesis pathway of citrulline, were studied in the diploid, triploid and tetraploid Yixuan watermelon. The results showed that the content of citrulline and ornithine in polyploid watermelon fruits were significantly higher than those in diploid, while the content of arginine in polyploid was lower than that in diploid. At 32 days of fruit development, the content of citrulline and ornithine were triploid ＞ tetraploid ＞ diploid, while the content of glutamic acid and arginine were diploid ＞ tetraploid ＞ triploid. Correspondingly, there were significant differences in the expression of NAGS, NAGK, GAT, OTC genes between polyploid and diploid in the late stage of fruit development, and the expression of ASS1, ASS3 and ARG genes in diploid was always higher than that of polyploid throughout the developmental period. These results indicated that the differences in citrulline content in different ploidy watermelon fruits were caused by the stable differential expression of upstream and downstream genes of citrulline synthesis pathway. This study is of great significance to reveal the mechanism of citrulline content difference in different ploidy watermelon.

关键词

西瓜 / 倍性 / 瓜氨酸 / 基因表达

Key words

Watermelon / Ploidy / Citrulline / Gene expression

参考文献

[1] GUO S G, ZHAO S J, SUN H H, et al. Resequencing of 414 cultivated and wild watermelon accessions identifies selection for fruit quality traits[J]. Nature genetics, 2019, 51(11): 1616-1623.

[2] TIAN S W, JIANG L J, GAO Q, et al. Efficient CRISPR/Cas9-based gene knockout in watermelon[J]. Plant Cell Reports, 2017, 36(3): 399-406.

[3] 刘启振, 王思明. 西瓜的起源及其在全球的传播[J]. 海交史研究, 2020(4): 30-44.

[4] 李福, 谢露露, 刘季扬, 等. 西瓜果实耐裂性的生理基础及其遗传和环境调控[J]. 中国瓜菜, 2023, 36(5): 1-7.

[5] UMER M J, BIN SAFDAR L, GEBRENESKEL H, et al. Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles[J]. Horticulture Research, 2020, 7(1): 193.

[6] 黄小忠, 卢阳月, 林远至, 等. 不同瓤色西瓜果肉挥发性物质检测与分析[J]. 中国瓜菜, 2023, 36(11): 33-39.

[7] CURIS E, NICOLIS I, MOINARD C, et al. Almost all about citrulline in mammals[J]. Amino Acids, 2005, 29(3): 177-205.

[8] COLLINS J K, WU G Y, PERKINS-VEAZIES P, et al. Watermelon consumption increases plasma arginine concentrations in adults[J]. Nutrition, 2007, 23(3): 261-266.

[9] JOSHI V, FEMIE A R. Citrulline metabolism in plants[J]. Amino Acids, 2017, 49(9): 1543-1559.

[10] BAHRI S, ZERROUK N, AUSSEL C, et al. Citrulline: From metabolism to therapeutic use[J]. Nutrition, 2013, 29(3): 479-484.

[11] 杨东东, 朱红菊, 赵永, 等. 蔬菜作物中瓜氨酸代谢调控研究进展[J]. 中国瓜菜, 2023, 36(2): 1-10.

[12] 朱红菊, 赵胜杰, 路绪强, 等. 二倍体和四倍体西瓜幼苗根系形态差异比较[J]. 中国瓜菜, 2020, 33(11): 18-21.

[13] COHEN H, FAIT A, TEL-ZUR N. Morphological, cytological and metabolic consequences of autopolyploidization in Hylocereus (Cactaceae) species[J]. BMC Plant Biology, 2013, 13(173): 2013.

[14] LAVANIA U C, SRIVASTAVA S, LAVANIA S, et al. Autopolyploidy differentially influences body size in plants, but facilitates enhanced accumulation of secondary metabolites, causing increased cytosine methylation[J]. Plant Journal, 2012, 71(4): 539-549.

[15] MENGIST M F, GRACE M H, XIONG J, et al. Diversity in metabolites and fruit quality traits in blueberry enables ploidy and species differentiation and establishes a strategy for future genetic studies[J]. Frontiers in Plant Science, 2020, 11: 370.

[16] CORNEILLIE S, DE STORME N, VAN ACKER R, et al. Polyploidy affects plant growth and alters cell wall composition[J]. Plant Physiology, 2019, 179(1): 74-87.

[17] 陆柳英, 阮孟斌, 吴正丹, 等. 蛋白质组学分析染色体加倍对木薯表型及块根形成期代谢产物的影响[J]. 植物生理学报, 2023(10): 1870-1880.

[18] TAN F Q, ZHANG M, XIE K D, et al. Polyploidy remodels fruit metabolism by modifying carbon source utilization and metabolic flux in Ponkan mandarin (Citrus reticulata Blanco)[J]. Plant Science, 2019, 289: 110276.

[19] 王友平, 朱金英, 徐光东, 等. 西瓜多倍体研究应用进展[J]. 长江蔬菜, 2010(20): 8-12.

[20] 刘文革, 王鸣, 阎志红. 蔬菜作物多倍体育种研究进展[J]. 长江蔬菜, 2003(1): 29-33.

[21] 郑锋, 詹园凤, 党选民, 等. 不同倍性小型西瓜果实中番茄红素、瓜氨酸含量比较分析[J]. 热带农业科学, 2012, 32(2): 68-73.

[22] 王学征, 张雅楠, 肖琪, 等. 西瓜果实发育过程中番茄红素积累变化及相关酶基因表达[J]. 东北农业大学学报, 2016, 47(4): 40-48.

[23] 万学闪, 刘文革, 阎志红, 等. 西瓜果实发育过程中番茄红素、瓜氨酸和维生素C等功能物质含量的变化[J]. 中国农业科学, 2011, 44(13): 2738-2747.

[24] VAN HIEU P. Polyploid gene expression and regulation in polysomic polyploids[J]. American Journal of Plant Sciences, 2019, 10(8): 1409-1443.

[25] IANNICELLI J, GUARINIELLO J, TOSSI V E, et al. The “polyploid effect” in the breeding of aromatic and medicinal species[J]. Scientia Horticulturae, 2019, 260: 108854.

[26] CORNEILLIE S, DE STPRME N, VAN ACKER R, et al. Polyploidy affects plant growth and alters cell wall composition[J]. Plant Physiology, 2019, 179(1): 74-87.

[27] TAN F Q, TU H, WANG R, et al. Metabolic adaptation following genome doubling in citrus doubled diploids revealed by non-targeted metabolomics[J]. Metabolomics, 2017, 13(11): 143.

[28] JOSHI V, JOSHI M, SILWAL D, et al. Systematized biosynthesis and catabolism regulate citrulline accumulation in watermelon[J]. Phytochemistry, 2019, 162: 129-140.

[29] AKASHI K, MIYAKE C, YOKOTA A. Citrulline, a novel compatible solute in drought-tolerant wild watermelon leaves, is an efficient hydroxyl radical scavenger[J]. Febs Letters, 2001, 508(3): 438-442.

[30] DASGAN H Y, KUSVURAN S, ABAK K, et al. The relationship between citrulline accumulation and salt tolerance during the vegetative growth of melon (Cucumis melo L.) [J]. Plant, Soil and Environment, 2009, 55(2): 51-57.

[31] 李蒙蒙, 路绪强, 赵胜杰, 等. 嫁接对西瓜果实瓜氨酸含量及合成途径关键酶基因表达的影响[J]. 果树学报, 2019, 36(7): 857-865.

[32] SONG Q S, JOSHI M, DIPIAZZA J, et al. Functional relevance of citrulline in the vegetative tissues of watermelon during abiotic stresses[J]. Frontiers in Plant Science, 2020, 11: 512.

基金资助

河南省科技攻关项目(242102110242)

中国农业科学院科技创新工程专项(CAAS-ASTIP-2021-ZFRI)

国家现代农业产业技术体系(CARS-25-03)

河南省重大科技专项(221100110400)

河南省农业良种联合攻关项目(2022010503)