盐碱胁迫对棉花叶片蛋白质组的影响及差异性分析

doi:10.3724/SP.J.1006.2024.34035

作物学报 ›› 2024, Vol. 50 ›› Issue (1): 219-236.doi: 10.3724/SP.J.1006.2024.34035

盐碱胁迫对棉花叶片蛋白质组的影响及差异性分析

郭家鑫(), 叶扬, 郭慧娟, 闵伟^*()

石河子大学农学院, 新疆石河子 832000

收稿日期:2023-02-22 接受日期:2023-05-24 出版日期:2024-01-12 网络出版日期:2023-12-16
通讯作者: *闵伟, E-mail: minwei555@126.com
作者简介:E-mail: gjx19960323@126.com
基金资助:
石河子大学青年创新人才培养计划项目(CXPY202111);农业农村部西北绿洲农业环境重点实验室开放基金(XBLZ-20214);石河子大学高层次人才科研启动资金专项(RCZK202017)

Effects and variability analysis of different salt and alkali stresses on the proteome of cotton leaves

GUO Jia-Xin(), YE Yang, GUO Hui-Juan, MIN Wei^*()

Agricultual College, Shihezi University, Shihezi 832000, Xinjiang, China

Received:2023-02-22 Accepted:2023-05-24 Published:2024-01-12 Published online:2023-12-16
Contact: *E-mail: minwei555@126.com
Supported by:
Youth Innovation Talent Cultivation Program of Shihezi University(CXPY202111);Open Fund of Key Laboratory of Northwest Oasis Agro-Environment of Ministry of Agriculture and Rural Affairs of China(XBLZ-20214);Scientific Research Starting Foundation for High Level Talents of Shihezi University(RCZK202017)

摘要/Abstract

摘要：

盐胁迫和碱胁迫是严重影响农业生产的2种不同非生物胁迫, 探讨棉花耐受盐胁迫和碱胁迫的差异, 可为不同类型盐碱地的棉花栽培提供一定的理论基础。本试验设置对照(CK)、盐胁迫(NaCl, CS)和碱胁迫(NaHCO₃+Na₂CO₃, AS) 3个处理, 通过蛋白质组学分析和生理指标进行验证, 揭示棉花对盐胁迫和碱胁迫的耐受机制。与CK相比, CS和AS的棉花生物量分别降低51.1%和50.9%, CS棉花叶片的叶绿素含量和净光合速率分别降低53.9%和57.2%, CS棉花叶片中己糖激酶、磷酸果糖激酶、丙酮酸激酶、柠檬酸合酶、谷氨酸脱氢酶和谷草转氨酶活性分别增加13.8%、14.4%、4.7%、4.5%、36.6%和12.9%; AS棉花叶片中己糖激酶、磷酸果糖激酶、丙酮酸激酶、苹果酸脱氢酶、柠檬酸合酶、谷氨酸脱氢酶和谷草转氨酶活性分别增加4.8%、38.8%、15.1%、4.3%、3.4%、15.2%和21.1%。基于TMT蛋白质组学分析, 在CS和AS叶片中分别鉴定出458个和140个差异表达蛋白质。这些蛋白参与了光合作用、糖代谢、2-氧羧酸代谢以及氨基酸合成和代谢等生命过程。表明盐胁迫和碱胁迫均抑制棉花生长。盐胁迫下, 与光合作用相关的蛋白表达减少, 光合作用被显著抑制, 同时碳水和能量代谢加强, 更多的光合产物用于能量代谢; 而碱胁迫对棉花光合作用无显著影响, 更多的光合产物可能运往根部分泌有机酸。

关键词: 盐胁迫, 碱胁迫, 棉花, 蛋白质组, 光合作用

Abstract:

Salt stress and alkali stress are two different abiotic stresses that seriously affect agricultural production. Exploring the differences between salt stress and alkali stress in cotton provides a theoretical basis for cotton cultivation on different types of saline alkali land. In this experiment, three treatments of CK, NaCl (CS), and NaHCO₃+Na₂CO₃ (AS) were conducted. Proteomics analysis and physiological index verification revealed that the tolerance mechanism of cotton to salt stress and alkali stress. Compared with CK, CS, and AS cotton plants decreased by 51.1% and 50.9% at P < 0.05 in biomass, respectively. The chlorophyll content and P_n under salt stress decreased significantly by 53.9% and 57.2%, respectively. Under salt stress, the activities of hexokinase, fructose phosphate kinase, pyruvate kinase, citrate synthase, glutamate dehydrogenase, and glutamic oxaloacetic transaminase in cotton leaves increased significantly by 13.8%, 14.4%, 4.7%, 4.5%, 36.6%, and 12.9%, respectively. Under alkali stress, the activities of hexokinase, phosphofructose kinase, pyruvate kinase, malate dehydrogenase, citrate synthase, glutamate dehydrogenase, and glutamic oxaloacetic transaminase in cotton leaves significantly increased by 4.8%, 38.8%, 15.1%, 4.3%, 3.4%, 15.2%, and 21.1%, respectively. Based on TMT proteomics, 458 differentially expressed proteins and 140 differentially expressed proteins in salt stress and alkali stress species were detected, respectively. These proteins were involved in photosynthesis, sugar metabolism, 2-oxocarbonic acid metabolism, amino acid synthesis and metabolism, and other life processes. These results showed that both salt stress and alkali stress inhibited cotton growth. The difference was that under salt stress, proteins expression related to photosynthesis decreased, photosynthesis was significantly inhibited, and carbon water and energy metabolism were enhanced, and more photosynthates were used for energy metabolism. Alkali stress had no significant effect on cotton photosynthesis, and more photosynthates might be transported to the roots to secrete organic acids.

Key words: salt stress, alkali stress, cotton, proteome, photosynthesis

郭家鑫, 叶扬, 郭慧娟, 闵伟. 盐碱胁迫对棉花叶片蛋白质组的影响及差异性分析[J]. 作物学报, 2024, 50(1): 219-236.

GUO Jia-Xin, YE Yang, GUO Hui-Juan, MIN Wei. Effects and variability analysis of different salt and alkali stresses on the proteome of cotton leaves[J]. Acta Agronomica Sinica, 2024, 50(1): 219-236.

图/表 13

表1

表2

图1

图2

图3

图4

图5

图6

表3

盐胁迫和碱胁迫对棉花叶片能量代谢的影响"

处理 Treatment	代谢通路 Pathway name	序列号 Accession	功能描述 Description	变化倍数 Fold change (CS or AS)/CK
CS	2-氧代羧酸代谢 2-oxocarboxylic acid metabolism	A0A7J9KP33	酮酸还原酶 Ketol-acid reductoisomerase	1.29^*
		A0A7J9MR64	乙酰谷氨酸激酶 Acetylglutamate kinase	1.36^**
		A0A7J9M5F7	乌头酸水合酶 Aconitate hydratase	0.73^*
		A0A7J9N0C5	芳香脱硫葡萄糖酸磺基转移酶 Aromatic desulfoglucosinolate sulfotransferase	1.27^*
	氧化磷酸化 Oxidative phosphorylation	A0A7J9LU75	F型H⁺转运ATP酶亚基δ F-type H⁺-transporting ATPase subunit delta	1.37^*
		A0A7J9LUR9	F型H⁺转运ATP酶亚基γ F-type H⁺-transporting ATPase subunit gamma	1.21^*
		A0A7J9KQM1	F型H⁺转运ATP酶亚基d F-type H⁺-transporting ATPase subunit d	1.57^*
		A0A7J9MB87	F型H⁺转运ATP酶亚基g F-type H⁺-transporting ATPase subunit g	1.40^**
		A0A7J9LJA3	V型H⁺转运ATP酶亚基D V-type H⁺-transporting ATPase subunit D	1.31^*
		A0A7J9N9Y3	细胞色素c氧化酶亚基5b Cytochrome c oxidase subunit 5b	2.86^**
		A0A7J9ML93	NADH脱氢酶(泛醌) 1α亚基1 NADH dehydrogenase (ubiquinone) 1 alpha subcomplex subunit 1	1.27^**
		A0A7J9LKQ6	NADH脱氢酶(泛醌)1α亚基9 NADH dehydrogenase (ubiquinone) 1 alpha subcomplex subunit 9	1.22^*
	脂肪酸代谢 Fatty acid metabolism	A0A7J9M364	长链酰基辅酶A合成酶 Long-chain acyl-CoA synthetase	1.68^**
		A0A7J9KWA3	3-羟基酰基-[酰基载体蛋白]脱水酶 3-hydroxyacyl-[acyl-carrier-protein] dehydratase	1.24^*
		A0A7J9LZQ8	3-氧代酰基-[酰基载体蛋白]合成酶II 3-oxoacyl-[acyl-carrier-protein] synthase II	1.29^*
	原核生物中的碳固定途径 Carbon fixation pathways in prokaryotes	A0A7J9KVG7	磷酸烯醇式丙酮酸羧化酶 Phosphoenolpyruvate carboxylase	0.64^*
	原核生物中的碳固定途径 Carbon fixation pathways in prokaryotes	A0A7J9M5F7	乌头酸水合酶 Aconitate hydratase	0.73^*
	甲烷代谢 Methane metabolism	A0A7J9KVG7	磷酸烯醇式丙酮酸羧化酶 Phosphoenolpyruvate carboxylase	0.64^*
	甲烷代谢 Methane metabolism	A0A7J9KMU6	烯醇化酶 Enolase	1.25^*
AS	2-氧代羧酸代谢 2-Oxocarboxylic acid metabolism	A0A7J9KP33	酮酸还原酶 Ketol-acid reductoisomerase	1.39^*
		A0A7J9M6Q9	支链氨基酸氨基转移酶 Branched-chain amino acid aminotransferase	0.83^**
		A0A7J9L982	乙酰乳酸合成酶I/III小亚基 Acetolactate synthase I/III small subunit	1.22^*
AS	脂肪酸代谢 Fatty acid metabolism	A0A7J9N2P9	酰基-[酰基载体蛋白]去饱和酶 Acyl-[acyl-carrier-protein] desaturase	1.39^*
	脂肪酸代谢 Fatty acid metabolism	A0A7J9LKB6	乙酰辅酶A酰基转移酶1 Acetyl-CoA acyltransferase 1	0.71^**
	硫代谢 Sulfur metabolism	A0A7J9MMD9	胱硫醚γ合酶 Cystathionine gamma-synthase	1.48^*
	硫代谢 Sulfur metabolism	A0A7J9L9D5	3’-磷酸腺苷5’-磷酸硫酸合成酶 3’-phosphoadenosine 5’-phosphosulfate synthase	1.29^*
	氧化磷酸化 Oxidative phosphorylation	A0A7J9LGN1	无机焦磷酸酶 Inorganic pyrophosphatase	0.80^*
	氧化磷酸化 Oxidative phosphorylation	A0A7J9LJA3	V型H⁺转运ATP酶亚基D V-type H⁺-transporting ATPase subunit D	1.26^**

表3

表4

盐胁迫和碱胁迫对棉花叶片氨基酸代谢的影响"

处理 Treatment	代谢通路 Pathway name	序列号 Accession	功能描述 Description	变化倍数 Fold change (CS or AS)/CK
CS	苯丙氨酸、酪氨酸和色氨酸生物合成 Phenylalanine, tyrosine, and tryptophan biosynthesis	A0A7J9LV20	蒽酰磷酸核糖转移酶 Anthranilate phosphoribosyl transferase	1.30^*
		A0A7J9KNK6	蒽合酶 Anthranilate synthase	0.83^**
		A0A7J9N2J2	色氨酸合成酶α链 Tryptophan synthase alpha chain	1.64^**
		A0A7J9N2X9	色氨酸合成酶β链 Tryptophan synthase beta chain	1.78^**
		A0A7J9N2E0	芳香烃脱水酶 Arogenate dehydratase	1.32^*
		A0A7J9LLN6	色氨酸合成酶β链 Tryptophan synthase beta chain	1.39^*
	甘氨酸、丝氨酸和苏氨酸代谢 Glycine, serine and threonine metabolism	A0A7J9N2J2	色氨酸合成酶α链 Tryptophan synthase alpha chain	1.64^**
		A0A7J9N2X9	色氨酸合成酶β链 Tryptophan synthase beta chain	1.78^**
		A0A7J9M6Y0	苏氨酸合酶 Threonine synthase	1.57^**
		A0A7J9LLN6	色氨酸合成酶β链 Tryptophan synthase beta chain	1.39^*
		A0A7J9M104	乙醛酸/羟基丙酮酸还原酶 Glyoxylate/hydroxypyruvate reductase	0.79^**
	酪氨酸代谢 Tyrosine metabolism	A0A7J9MAI5	多酚氧化酶 Polyphenol oxidase	0.68^*
		A0A7J9M832	富马酸酯酶 Fumarylacetoacetase	0.75^*
		A0A7J9M104	羟基苯基丙酮酸还原酶 Hydroxyphenylpyruvate reductase	0.79^**
	精氨酸生物合成 Arginine biosynthesis	A0A7J9MR64	乙酰谷氨酸激酶 Acetylglutamate kinase	1.36^**
CS	精氨酸生物合成 Arginine biosynthesis	A0A7J9KRZ6	精氨酸琥珀酸裂解酶 Argininosuccinate lyase	1.53^*
	丙氨酸、天冬氨酸和谷氨酸代谢 Alanine, aspartate, and glutamate metabolism	A0A7J9KPZ3	谷氨酸脱羧酶 Glutamate decarboxylase	0.77^*
		A0A7J9KRZ6	精氨酸琥珀酸裂解酶 Argininosuccinate lyase	1.53^*
	色氨酸代谢 Tryptophan metabolism	A0A7J9KNP8	细胞色素P450家族1亚家族A1 Cytochrome P450 family 1 subfamily A1	0.16^**
	色氨酸代谢 Tryptophan metabolism	A0A7J9N0C5	芳香脱硫葡萄糖酸磺基转移酶 Aromatic desulfoglucosinolate sulfotransferase	1.27^*
AS	缬氨酸、亮氨酸和异亮氨酸生物合成 Valine, leucine, and isoleucine biosynthesis	A0A7J9KP33	酮酸还原酶 Ketol-acid reductoisomerase	1.39^*
		A0A7J9M6Q9	支链氨基酸氨基转移酶 Branched-chain amino acid aminotransferase	0.83^*
		A0A7J9L982	乙酰乳酸合成酶I/III小亚基 Acetolactate synthase I/III small subunit	1.22^*
	苯丙氨酸、酪氨酸和色氨酸生物合成 Phenylalanine, tyrosine, and tryptophan biosynthesis	A0A7J9LV20	蒽酰磷酸核糖转移酶 Anthranilate phosphoribosyl transferase	1.25^*
		A0A7J9N2J2	色氨酸合成酶α链 Tryptophan synthase alpha chain	1.41^*
		A0A7J9N2X9	色氨酸合成酶β链 Tryptophan synthase beta chain	1.49^*
		A0A7J9MCV6	预苯酸脱水酶 Prephenate dehydratase	1.29^**
		A0A7J9M850	芳香酸脱氢酶(NADP⁺) Arogenate dehydrogenase (NADP⁺)	1.44^*
	缬氨酸、亮氨酸和异亮氨酸降解 Valine, leucine, and isoleucine degradation	A0A7J9M6Q9	支链氨基酸氨基转移酶 Branched-chain amino acid aminotransferase	0.83^*
		A0A7J9LT42	羟甲基戊二酰辅酶A合酶 Hydroxymethylglutaryl-CoA synthase	1.27^**
		A0A7J9LKB6	乙酰辅酶A酰基转移酶1 Acetyl-CoA acyltransferase 1	0.71^*
	半胱氨酸和蛋氨酸代谢 Cysteine and methionine metabolism	A0A7J9LQ81	5-甲基四氢蝶酰三谷氨酸 5-methyltetrahydropteroyltriglutamate	0.73^*
		A0A7J9M6Q9	支链氨基酸氨基转移酶 Branched-chain amino acid aminotransferase	0.83^*
		A0A7J9MMD9	胱硫苷γ合酶 Cystathionine gamma-synthase	1.48^*
		A0A7J9LCJ0	1,2-二羟基-3-酮-5-甲基硫戊烯双加氧酶 1,2-dihydroxy-3-keto-5-methylthiopentene dioxygenase	0.80^**
	甘氨酸、丝氨酸和苏氨酸代谢 Glycine, serine, and threonine metabolism	A0A7J9N2J2	色氨酸合成酶α链 Tryptophan synthase alpha chain	1.41^*
	甘氨酸、丝氨酸和苏氨酸代谢 Glycine, serine, and threonine metabolism	A0A7J9N2X9	色氨酸合成酶β链 Tryptophan synthase beta chain	1.49^*

表4

图7

图8

表5

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处理 Treatment	盐碱类型及盐碱化程度 Saline and alkaline	含盐量 Salt content (g kg^-1)	电导率 Electrical conductivity EC_1:5(dS m^-1)	pH (1:2.5)
对照 CK	对照-非盐(碱)化 Control-non salting (alkalization)	0.53	0.17	8.16
盐胁迫 CS	NaCl-中度盐化 NaCl-moderate salinization	4.43	1.39	8.43
碱胁迫 AS	Na₂CO₃+NaHCO₃-中度碱化 Na₂CO₃+NaHCO₃-moderate alkalization	2.03	0.63	9.92

处理 Treatment	生物量Biomass (g plant^-1)
处理 Treatment	叶Leaf	茎Stem	根Root	总Total
CK	2.45±0.200 a	1.81±0.100 a	0.87±0.040 a	5.13±0.260 a
CS	1.24±0.015 b	0.73±0.006 c	0.54±0.015 c	2.51±0.025 c
AS	1.27±0.095 b	0.72±0.050 b	0.53±0.045 b	2.52±0.190 b

处理 Treatment	代谢通路 Pathway name	序列号 Accession	功能描述 Description	变化倍数 Fold change (CS or AS)/CK
CS	AMPK信号通路 AMPK signaling pathway	A0A7J9L324	5'-AMP激活蛋白激酶, 调节β亚基 5'-AMP-activated protein kinase, regulatory beta subunit	1.29^*
		A0A7J9MK06	5'-AMP激活蛋白激酶, 调节γ亚基 5'-AMP-activated protein kinase, regulatory gamma subunit	0.64^**
		A0A7J9MBY1	钙结合蛋白39 Calcium binding protein 39	1.37^*
	MAPK信号通路 MAPK signaling pathway	A0A7J9KIX8	酪蛋白激酶1 Casein kinase 1	0.75^*
	MAPK信号通路 MAPK signaling pathway	A0A7J9KNP5	14-3-3蛋白ε 14-3-3 protein epsilon	1.45^*
	FoxO信号通路 FoxO signaling pathway	A0A7J9L324	5'-AMP激活蛋白激酶, 调节β亚基 5'-AMP-activated protein kinase, regulatory beta subunit	1.29^*
	FoxO信号通路 FoxO signaling pathway	A0A7J9MK06	5'-AMP激活蛋白激酶, 调节γ亚基 5'-AMP-activated protein kinase, regulatory gamma subunit	0.64^**
	Apelin信号通路 Apelin signaling pathway	A0A7J9L324	5'-AMP激活蛋白激酶, 调节β亚基 5'-AMP-activated protein kinase, regulatory beta subunit	1.29^*
	Apelin信号通路 Apelin signaling pathway	A0A7J9MK06	5'-AMP激活蛋白激酶, 调节γ亚基 5'-AMP-activated protein kinase, regulatory gamma subunit	0.64^**
CS	mTOR信号通路 mTOR signaling pathway	A0A7J9LJA3	V型H⁺转运ATP酶亚基D V-type H⁺-transporting ATPase subunit D	1.31^*
CS	mTOR信号通路 mTOR signaling pathway	A0A7J9MBY1	钙结合蛋白39 Calcium binding protein 39	1.37^*
AS	植物激素信号转导 Plant hormone signal transduction	A0A7J9LIK5	含组氨酸磷酸转移蛋白 Histidine-containing phosphotransfer peotein	1.30^**
	植物激素信号转导 Plant hormone signal transduction	A0A7J9MIA1	丝氨酸/苏氨酸蛋白激酶SRK2 Serine/threonine-protein kinase SRK2	1.23^*
	mTOR信号通路 mTOR signaling pathway	A0A7J9LJA3	V型H⁺转运ATP酶亚基D V-type H⁺-transporting ATPase subunit D	1.26^**
	mTOR信号通路 mTOR signaling pathway	A0A7J9MBY1	钙结合蛋白39 Calcium binding protein 39	1.29^*

盐碱胁迫对棉花叶片蛋白质组的影响及差异性分析

Effects and variability analysis of different salt and alkali stresses on the proteome of cotton leaves

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