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作物学报

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

• 耕作栽培·生理生化 • 上一篇    下一篇

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*()   

  1. 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*()   

  1. 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)

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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 Pn 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

表1

不同处理土壤盐碱类型和盐碱化程度"

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 EC1:5 (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

表2

盐胁迫和碱胁迫对棉花生物量的影响"

处理
Treatment		 生物量Biomass (g plant-1)
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

图1

盐胁迫和碱胁迫对棉花光合作用的影响 柱上不同小写字母表示不同处理在0.05概率水平差异显著。处理同表2。"

图2

盐胁迫和碱胁迫下棉花叶片中表达蛋白的主坐标分析 处理同表2。"

图3

棉花叶片中表达蛋白火山图 A: CS vs CK; B: AS vs CK。散点颜色: 显著上调的蛋白质以红色表示, 显著下调的蛋白质以蓝色表示, 非显著差异的蛋白质为灰色。处理同表2。"

图4

棉花叶片差异表达蛋白GO富集分析分类直方图 A: CS vs CK; B: AS vs CK。富集条目为top 10, 纵坐标为映射差异表达蛋白个数。BP: 生物过程; CC: 细胞组分; MF: 分子功能。处理同表2。"

图5

棉花叶片差异表达蛋白的前十位KEGG代谢通路富集分析图 A: CS vs CK; B: AS vs CK。图中横坐标为富集因子值, 纵坐标为KEGG 通路信息。其中, 圆圈的大小表示映射通路的差异表达蛋白个数, 圆圈越大, 数量越多; 圆圈的颜色表示P值大小, 颜色越红, P值越小。处理同表2。"

图6

盐胁迫和碱胁迫对棉花光合作用和碳转化蛋白的影响 A: 光合作用; B: 糖酵解; C: 柠檬酸循环; D: 氨基酸转化。柱上不同小写字母表示不同处理在0.05概率水平差异显著。处理同表2。"

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*
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*
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*
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*
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*
A0A7J9LJA3 V型H+转运ATP酶亚基D
V-type H+-transporting ATPase subunit D		 1.26**

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**
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*
A0A7J9N2X9 色氨酸合成酶β链
Tryptophan synthase beta chain		 1.49*

图7

盐胁迫对蛋白代谢的影响 A: 翻译过程; B: 复制和修复过程; C: 转录过程; D: 折叠、分类和降解过程。处理同表2。"

图8

碱胁迫对蛋白代谢的影响 A: 转录过程; B: 折叠、分类和降解过程; C: 翻译过程。处理同表2。"

表5

盐胁迫和碱胁迫对棉花叶片信号传导的影响"

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 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*
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*
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*
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*
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**
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**
A0A7J9MBY1 钙结合蛋白39
Calcium binding protein 39		 1.29*
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