茉莉酸调控马铃薯离体块茎发育的主要代谢物变化

doi:10.3864/j.issn.0578-1752.2024.13.003

摘要/Abstract

摘要：

【目的】茉莉酸（jasmonic acid，JA）是马铃薯块茎发育过程中重要调控激素之一，研究JA调控块茎发育机理将为块茎产量及品质性状形成提供重要理论基础。【方法】采用马铃薯离体匍匐茎作为供试材料，外源添加JA（0、0.5、5和50 μmol·L^-1）处理，分析JA诱导离体块茎表型形态、组织显微结构、碳水化合物积累及蛋白质组变化。【结果】随JA处理浓度升高，单个匍匐茎形成块茎数目、块茎直径及干鲜重、环髓区细胞面积、淀粉及可溶性糖含量在0.5和5 μmol·L^-1 JA处理后逐渐增加（P<0.05），而50 μmol·L^-1 JA处理后块茎直径及干鲜重、淀粉含量显著减少（P<0.05）；脂氧合酶活性随JA浓度升高逐渐降低（P<0.05）。采用双向凝胶电泳（2-DE）和MALDI-TOF/TOF-MS质谱技术鉴定到JA调控块茎发育密切相关的35个差异表达蛋白质（P<0.05且差异表达倍数≥2.5倍），主要涉及生物能量与代谢（28.6%）、细胞防御（28.6%）、蛋白质生物合成与贮藏（11.4%）、信号转导（8.6%）、转录（8.6%）、未知功能（8.6%）和混杂（5.6%）。通过聚类分析将这些蛋白质差异表达模式聚为三类：第一类包括17个蛋白质，在0.5 μmol·L^-1 JA处理后下调表达，而在5 μmol·L^-1 JA处理后上调表达，主要参与生物能量与代谢、蛋白质生物合成与贮藏、信号转导、转录；第二类包括10个蛋白质，随JA浓度升高上调表达，主要参与细胞防御、生物能量与代谢、转录；第三类包括8个蛋白质，JA处理后均下调表达，主要参与生物能量与代谢、细胞防御、蛋白质生物合成与贮藏。【结论】低浓度JA（0.5和5 μmol·L^-1）主要通过诱导块茎环髓区细胞膨大、细胞内蔗糖及多糖积累、细胞防御能力来促进块茎形态建成，而高浓度JA（50 μmol·L^-1）则表现为抑制作用。

关键词: 马铃薯, 茉莉酸, 蛋白质组, 块茎发育, 脂氧合酶, 质谱

Abstract:

【Objective】 Jasmonic acid (JA) is one of key regulatory hormones during potato tuber development. Study on JA regulating tuber development mechanism would provide an important theoretical basis for tuber yield and quality formation.【Method】 In this study, potato stolons cultured in vitro were exogenously applied with JA (0, 0.5, 5, 50 μmol·L^-1) to analyze the phenotype, tissue microstructure, carbohydrate accumulation and proteome change of tubers.【Result】 With the increase of JA concentration, the tuber number formed per stolon, tuber diameter, fresh and dry weight, cell area of perimedullary zone, starch and soluble sugar content were gradually increased at 0.5 and 5 μmol·L^-1 JA (P<0.05), whereas the tuber diameter, fresh and dry weight, starch content were significantly decreased at 50 μmol·L^-1 JA (P<0.05). Lipoxygenase activity was gradually decreased with JA concentration increasing (P<0.05). Thirty-five differentially abundant proteins (P<0.05 and fold change≥2.5) closely related to tuber development regulating by JA were identified by 2-DE and MALDI-TOF/TOF-MS. They were mainly involved in bioenergy and metabolism (28.6%), cell defense and rescue (28.6%), protein biogenesis and storage (11.4%), signaling (8.6%), transcription (8.6%), unknown (8.6%) and miscellaneous (5.6%). The differential expression patterns of these proteins were clustered into three categories by hierarchical clustering analysis. Cluster 1 included 17 proteins that down-regulated at 0.5 μmol·L^-1 JA and up-regulated at 5 μmol·L^-1 JA, which mainly involved in bioenergy and metabolism, protein biogenesis and storage, signaling and transcription. Cluster 2 included 10 proteins that gradually up-regulated with JA concentration increasing, which were mainly involved in cell defense and rescue, bioenergy and metabolism, transcription. Cluster 3 included 8 proteins that down-regulated after JA treatments, which were mainly involved in bioenergy and metabolism, cell defense and rescue, protein biogenesis and storage. 【Conclusion】 Low JA concentration (0.5, 5 μmol·L^-1) might promote tuber morphogenesis by inducing cell enlargement in tuber perimedullary zone, intracellular sucrose and polysaccharide accumulation, and cell defense ability, whereas high JA concentration (50 μmol·L^-1) showed inhibitory effects.

Key words: Solanum tuberosum, jasmonic acid, proteome, tuber development, lipoxygenase, mass spectrum

梁丽娟, 程李香, 袁剑龙, 撒刚, 张峰. 茉莉酸调控马铃薯离体块茎发育的主要代谢物变化[J]. 中国农业科学, 2024, 57(13): 2525-2538.

LIANG LiJuan, CHENG LiXiang, YUAN JianLong, SA Gang, ZHANG Feng. Jasmonic Acid Regulates the Changes of Major Metabolites in Potato Tuber Development in vitro[J]. Scientia Agricultura Sinica, 2024, 57(13): 2525-2538.

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JA调控马铃薯离体块茎发育的差异表达蛋白质鉴定结果"

点序号^a Spot No.	蛋白名称 Ptotein name	物种 Organism	gi号^b gi No.	理论等电点/相对分子质量^c Theoretical pI/Mr (kDa)	得分^d Score	匹配肽段^e PN
生物能量与代谢相关蛋白质Bioenergy and metabolism-related proteins
6606	3-磷酸甘油醛脱氢酶 Glyceraldehyde 3-phosphatedehydrogenase	Solanum tuberosum	gi\|22094840	6.34/36.79	882	17
4805	烯醇化酶 Enolase	Solanum lycopersicum	gi\|350538295	5.68/48.05	1120	23
3819	烯醇化酶 Enolase	Solanum lycopersicum	gi\|350538295	5.68/48.05	1070	15
7610	假定的线粒体NAD依赖的苹果酸脱氢酶 Putative mitochondrial NAD-dependent malate dehydrogenase	Solanum tuberosum	gi\|21388544	8.87/36.40	1000	15
1409	尿苷二磷酸葡萄糖醛基转移酶1-6 UDP-glucuronosyltransferase1-6	Medicago truncatula	gi\|357512853	6.20/54.68	45	7
7701	甲酸脱氢酶 Formate dehydrogenase	Solanum lycopersicum	gi\|350538487	6.87/42.35	864	19
0804	类3-酮脂酰-CoA合酶17 3-ketoacyl-CoA synthase 17-like	Vitis vinifera	gi\|359479575	8.24/52.53	59	10
1309	双半乳糖二酰基甘油合酶2 Digalactosyldiacylglycerol synthase 2	Brachypodium distachyon	gi\|357112938	8.65/53.57	40	8
7311	类细胞色素P450 86A1 Cytochrome P450 86A1-like	Glycine max	gi\|356553060	9.30/59.44	41	4
1804	细胞色素P450 Cytochrome P450	Arabidopsis thaliana	gi\|19698839	8.92/58.80	58	12
细胞防御相关蛋白质Cell defense and rescue-related proteins
1715	铜锌超氧化物歧化酶 Cu/Zn-superoxide dismutase	Solanum tuberosum	gi\|13751866	5.28/14.56	553	8
8711	过氧化物酶 Peroxidase	Nicotiana tabacum	gi\|14031049	5.99/39.49	440	12
5306	脱氢抗坏血酸还原酶 Dehydroascorbate reductase	Solanum tuberosum	gi\|160347100	6.09/23.61	653	12
6827	多酚氧化酶 Polyphenol oxidase	Solanum tuberosum	gi\|1146426	7.18/68.10	492	19
5822	多酚氧化酶 Polyphenol oxidase	Solanum tuberosum	gi\|1146426	7.18/68.10	379	15
5501	I型几丁质酶 Class I chitinase	Solanum tuberosum	gi\|6707113	5.66/36.33	494	6
7505	内切几丁质酶2 Endochitinase 2	Solanum tuberosum	gi\|1705808	5.94/34.57	685	8
8307	假定的无毒性响应蛋白 Putative avirulence-responsive protein	Arabidopsis thaliana	gi\|15217702	8.23/29.08	49	8
2302	核苷酸结合位点富含亮氨酸重复序列抗病蛋白Nucleotide binding site-leucine-rich repeat disease resistance protein	Pyrus communis	gi\|40644864	5.03/5.80	46	4
7112	羧酸酯酶 Carboxylesterase	Malus pumila	gi\|82697935	6.02/34.62	35	2
转录相关蛋白质Transcription-related proteins
6714	核RNA结合蛋白 Nuclear RNA binding protein	Solanum tuberosum	gi\|257215078	6.47/39.06	590	14
0102	假定的富含甘氨酸RNA结合蛋白 Putative glycine-rich RNA binding protein-like	Solanum tuberosum	gi\|82623423	5.58/17.56	599	12
3310	成熟酶K Maturase K	Ledermanniella pusilla	gi\|327204748	9.70/37.54	45	7
蛋白质生物合成与贮藏相关蛋白质Protein biogenesis and storage-related proteins
8001	蛋白酶抑制剂Ⅱ前体类型B Proteinase inhibitor Ⅱ precursor type B	Solanum tuberosum	gi\|334898813	5.69/17.60	447	10
9703	类延伸因子1α蛋白 Elongation factor 1-alpha-like protein	Solanum tuberosum	gi\|77999257	9.20/44.33	303	7
0009	类天冬氨酸蛋白酶前体 Aspartic protease precursor-like	Solanum tuberosum	gi\|82623417	5.85/55.71	420	8
1907	热激蛋白70 Heat shock 70 protein	Spinacia oleracea	gi\|2654208	5.19/76.26	885	18
信号转导相关蛋白质Signaling-related proteins
8108	丝裂原活化蛋白激酶 MAP kinase	Medicago truncatula	gi\|357437463	5.43/103.11	62	14
6416	含EF-手型结构域的钙结合蛋白 Calcium-binding EF-hand-containing protein	Arabidopsis thaliana	gi\|18407118	4.34/15.32	52	7
7320	类磷脂酰肌醇-4-磷酸5-激酶1 Phosphatidylinositol-4-phosphate 5-kinase 1-like	Brachypodium distachyon	gi\|357117945	8.74/81.57	46	11
混杂Miscellaneous
8203	果实成熟蛋白 Fruit-ripening protein	Solanum lycopersicum	gi\|350539427	6.48/12.54	927	13
1507	Ran结合蛋白-1 Ran binding protein-1	Solanum lycopersicum	gi\|350538193	4.88/25.22	665	9
未知功能Unknown
2203	假定蛋白OsJ_24584 Hypothetical protein OsJ_24584	Oryza sativa Japonica Group	gi\|222637200	8.24/69.08	60	10
2401	Os09g0491852 Os09g0491852	Oryza sativa Japonica Group	gi\|297727117	4.96/39.15	51	5
6310	未知Unknown	Picea sitchensis	gi\|294461039	6.51/8.11	45	3

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