巴西橡胶树6个PP2C家族基因成员分子信息学与抗旱功能分析
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摘要
PP2C是一类丝氨酸/苏氨酸残基蛋白磷酸酶,在高等植物ABA信号途径中起着重要的作用。为阐明巴西橡胶树中PP2C基因的结构与功能,本研究通过生物信息学方法,从橡胶树转录组数据库中鉴定并获得6个PP2C家族基因,均含有PP2CD、F1和F2亚族。通过qRT-PCR技术对6个PP2C家族基因进行了干旱处理下的差异表达分析,发现6个基因都不同程度上响应橡胶树干旱胁迫。本研究为探究PP2C基因在橡胶树抗干旱反应机制提供了理论依据。
PP2Cs are a group of monomeric serine/threonine protein phosphatases,which play an important role in ABA signaling pathway in higher plants. In order to elucidate the structure and function of PP2C gene in rubber tree,we used a bioinformatics approach to get six PP2C genes members from rubber tree transcription database,six HbPP2Cs all contain PP2C conserved domains. By sequence alignment and phylogenetic analysis,six HbPP2Cs were divided into four subfamilies: subfamily A,D,F1 and F2. Different expression analysis of six PP2C family genes under drought stress by qRT-PCR showed that they all response to drought stress in varying degrees. Our study provides a theoretical basis for exploring the mechanism of PP2C genes in drought resistance of rubber tree.
PP2Cs are a group of monomeric serine/threonine protein phosphatases,which play an important role in ABA signaling pathway in higher plants. In order to elucidate the structure and function of PP2C gene in rubber tree,we used a bioinformatics approach to get six PP2C genes members from rubber tree transcription database,six HbPP2Cs all contain PP2C conserved domains. By sequence alignment and phylogenetic analysis,six HbPP2Cs were divided into four subfamilies: subfamily A,D,F1 and F2. Different expression analysis of six PP2C family genes under drought stress by qRT-PCR showed that they all response to drought stress in varying degrees. Our study provides a theoretical basis for exploring the mechanism of PP2C genes in drought resistance of rubber tree.
引文
1.高伟,韦家少,何鹏,等.巴西橡胶树水解酶基因克隆与表达分析[J].中国农学通报,2013,29(19):9-13.Gao W,Wei G S,He P,et al.Cloning and Expression Analysis of Hydrolase Gene from Hevea brasiliensis Muell.Arg[J].Chinese Agricultural Science Bulletin,2013,29(19):9-13.
2 .钟勇.海南橡胶种植业发展前景分析[J].热带农业科学,2016,36(8):69-75.Zhong Y.Development Prospects of Rubber Industry in Hainan Province[J].Chinese Journal of Tropical Agriculture,2016,36(8):69-75.
3 .李春鸾,陈丽英,郑亚娜天.海南岛暴雨统计分析[J].气象研究与应用,2008,29:57-58.Li C Y,Chen L Y,Zheng Y N.Statistical analysis of Rainstorm in Hainan Island[J].Journal of Meteorological Research and Application,2008,29:57-58.
4 .李海亮,戴声佩,胡盛红,等.海南岛农业干旱综合监测研究[J].中山大学学报:自然科学版,2012,51(6):126-130.Li H L,Dai S P,Hu S H,et al.Study on Agricultural Drought Comprehensive Monitoring in Hainan Island Based on MODIS Data and Meteorological Data[J].Acta Scientiarum Naturalium Universitatis Sunyatsenii,2012,51(6):126-130.
5 .Kumagai T O,Mudd R G,Giambelluca T W,et al.How do rubber(Hevea brasiliensis)plantations behave under seasonal water stress in northeastern Thailand and central Cambodia?[J].Agricultural and Forest Meteorology,2015,213:10-22.
6 .Li Y W,Lan G Y,Xia Y J.Rubber trees demonstrate a clear retranslocation under seasonal drought and coldstresses[J].Frontiers in Plant Science,2016,7(e69357):1907.
7 .林秀琴,袁坤,王真辉,等.干旱胁迫下橡胶树叶片差异表达蛋白的鉴定与功能解析[J].热带作物学报,2009,30(12):1782-1788.Lin X Q,Yuan K,Wang Z H,et al.Identification and Functional Analysis of DIfferentially Expressed Proteins in Hevea brasiliensis Leaves during Drought Stress[J].Chinese Journal of Tropical Crops,2009,30(12):1782-1788.
8 .Wang L F.Physiological and molecular responses to drought stress in rubber tree(Hevea brasiliensis Muell.Arg.)[J].Plant Physiol and Biochemistry,2014,83(2):243-249.
9 .Xue T T,Wang D,Zhang S Z,et al.Genome-wide and expression analysis of protein phosphatase 2C in rice and arabidopsis[J].BMC Genomics,2008,9(1):550.
10 .Landi S,Nurcato R,De Lillo A,et al.Glucose-6-phosphate dehydrogenase plays a central role in the response of tomato(Solanum lycopersicum)plants to short and long-term drought[J].Plant Physiology and Biochemistry,2016,105:79-89.
11 .Zhang F Y,Fu X Q,LüZ Y,et al.Type 2C phosphatase 1of Artemisia annua L.is a negative regulator of ABA signaling[J].Bio Med Research International,2014,2014:521794.
12 .Wei K F,Pan S.Maize protein phosphatase gene family:identification and molecular characterization[J].BMC Genomics,2014,15(1):773.
13 .Stern A,Privman E,Rasis M,et al.Evolution of the metazoan protein phosphatase 2C superfamily[J].Journal of Molecular Evolution,2007,64(1):61-70.
14 .Xing Y N,Xu Y H,Chen Y,et al.Structure of protein phosphatase 2A core enzyme bound to tumor-inducing toxins[J].Cell,2006,127(2):341-353.
15 .Fuchs S,Grill E,Meskiene I,et al.Type 2C protein phosphatases in plants[J].FEBS Journal,2013,280(2):681-693.
16 .Cao J M,Jiang M,Li P,et al.Genome-wide identification and evolutionary analyses of the PP2C gene family with their expression profiling in response to multiple stresses in Brachypodium distachyon[J].BMC Genomics,2016,17(1):175.
17 .阮海华,徐朗莱.2C类蛋白磷酸酶的结构与功能研究进展[J].南京农业大学学报,2007,30(1):136-141.Ruan H H,Xu L L.Progress in the structure and function of PP2C type protein phosphatases[J].Journal of Nanjing Agricultural University,2007,30(1):136-141.
18 .胡晓丽,李德全.植物蛋白磷酸酶2C(PP2C)及其在信号转导中的作用[J].植物生理学通讯,2007,43(3):407-412.Hu X L,Li D Q.Protein Phosphatase 2C in Plants and Its Functions of Signal Transduction[J].Plant Physiology Communications,2007,43(3):407-412.
19 .Chen J H,Zhang D Z,Zhang C,et al.A putative PP2C-encoding gene negatively regulates ABA Signaling in Populus euphratica[J].PLo S One,2015,10(10):e0139466.
20 .Cui M H,Yoo K S,Hyoung S,et al.An Arabidopsis R2R3-MYB transcription factor,At MYB20,negatively regulates type 2C serine/threonine protein phosphatases to enhance salt tolerance[J].FEBS Letters,2013,587(12):1773-1778.
21 .Leung J,Merlot S,Giraudat J.The Arabidopsis ABSCISIC ACID-INSENSITIVE2(ABI2)and ABI1genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction[J].The Plant Cell,1997,9(5):759-771.
22 .Gosti F,Beaudoin N,Serizet C,et al.ABI1 protein phosphatase 2C is a negative regulator of abscisic acid signaling[J].The Plant Cell,1999,11(10):1897-1909.
23 .Schweighofer A,Kazanaviciute V,Scheikl E,et al.The PP2C-type phosphatase AP2C1,which negatively regulates MPK4 and MPK6,modulates innate immunity,jasmonic acid,and ethylene levels in Arabidopsis[J].The Plant Cell,2007,19(7):2213-2224.
24 .Meskiene I,Baudouin E,Schweighofer A,et al.Stress-induced protein phosphatase 2C is a negative regulator of a mitogen-activated protein kinase[J].Journal of Biological Chemistry,2003,278(21):18945-18952.
25 .郭鹏,张士刚,邢鑫,等.欧美杨Pd PP2C基因的克隆与功能分析[J].北京林业大学学报,2015,37(2):100-106.Guo P,Zhang S G,Xing X,et al.Cloning and Function analysis of Pd PP2C from Popululus deltoises[J].Journal of Beijing Forestry University,2015,37(2):100-106.
26 .闵东红,薛飞洋,马亚男,等.谷子PP2C基因家族的特性[J].作物学报,2013,39(12):2135-2144.Min D H,Xue F Y,Ma Y N,et al.Characteristics of PP2C Gene Family in Foxtail Millet(Setaria italica)[J].Acta Agronomica Sinica,2013,39(12):2135-2144.
27 .Vurukonda S S K P,Vardharajula S,Shrivastava M,et al.Enhancement of drought stress tolerance in crops by plant growth promoting rhizobacteria[J].Microbiological Research,2016,184:13-24.
28 .Basu S,Ramegowda V,Kumar A,et al.Plant adaptation to drought stress[J].F1000Research,2016,5:1554.
29 .Chaves M M,Pereira J S,Maroco J,et al.How plants cope with water stress in the field?Photosynthesis and growth[J].Annals of Botany,2002,89(7):907-916.
30 .李国尧,王权宝,李玉英,等.橡胶树产胶量影响因素[J].生态学杂志,2014,33(2):510-517.Li G Y,Wang Q B,Li Y Y,et al.A Review of Influencing Factors on Latex,Yeild of Hevea brasiliensis[J].Chinese Journal of Ecology,2014,33(2):510-517.
31 .Xu Z Z,Zhou G S,Shimizu H.Plant responses to drought and rewatering[J].Plant Signaling&Behavior,2010,5(6):649-654.
32 .Rodriguez P L.Protein phosphatase 2C(PP2C)function in high plants[J].Plant Molecular Biology,1998,38(6):919-927.
33 .Moorhead G B G,Trinkle-mulcahy L,Ulke-lemée A.Emerging roles of nuclear protein phosphatases[J].Nature Reviews Molecular Cell Biology,2007,8(3):234-244.
34 .Bhalothia P,Sangwan C,Alok A,et al.PP2C-like promoter and its deletion variants are induced by ABA but not by Me JA and SA in Arabidopsis thaliana[J].Frontiers in Plant Science,2016,7:547.
35 .Thomashow M F.Plant cold acclimation:freezing tolerance genes and regulatory mechanisms[J].Annual Review of Plant Physiology and Plant Molecular Biology,1999,50:571-599.
36 .Thtiharju S,Palva T.Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana[J].Plant Journal for Cell&Molecular Biology,2001,26(4):461-470.
37 .何亮,李富华,沙莉娜,等.玉米2C型丝氨酸/苏氨酸蛋白磷酸酶(PP2C)活性与耐旱性的关系[J].作物学报,2008,34(5):899-903.He L,Li F H,Sha Li Na,et al.Activity of Serine/Threonine Protein Phosphatase Type-2C(PP2C)and Its Relationships to Drought Tolerance in Maize[J].Acta Agronomica Sinica,2008,34(5):899-903.
38 .Miyazaki S,Koga R,Bohnert H J,et al.Tissue-and environmental response-specific expression of 10 PP2C transcripts in Mesembryanthemum crystallinum[J].Molecular and General Genetics MGG,1999,261(2):307-316.
2 .钟勇.海南橡胶种植业发展前景分析[J].热带农业科学,2016,36(8):69-75.Zhong Y.Development Prospects of Rubber Industry in Hainan Province[J].Chinese Journal of Tropical Agriculture,2016,36(8):69-75.
3 .李春鸾,陈丽英,郑亚娜天.海南岛暴雨统计分析[J].气象研究与应用,2008,29:57-58.Li C Y,Chen L Y,Zheng Y N.Statistical analysis of Rainstorm in Hainan Island[J].Journal of Meteorological Research and Application,2008,29:57-58.
4 .李海亮,戴声佩,胡盛红,等.海南岛农业干旱综合监测研究[J].中山大学学报:自然科学版,2012,51(6):126-130.Li H L,Dai S P,Hu S H,et al.Study on Agricultural Drought Comprehensive Monitoring in Hainan Island Based on MODIS Data and Meteorological Data[J].Acta Scientiarum Naturalium Universitatis Sunyatsenii,2012,51(6):126-130.
5 .Kumagai T O,Mudd R G,Giambelluca T W,et al.How do rubber(Hevea brasiliensis)plantations behave under seasonal water stress in northeastern Thailand and central Cambodia?[J].Agricultural and Forest Meteorology,2015,213:10-22.
6 .Li Y W,Lan G Y,Xia Y J.Rubber trees demonstrate a clear retranslocation under seasonal drought and coldstresses[J].Frontiers in Plant Science,2016,7(e69357):1907.
7 .林秀琴,袁坤,王真辉,等.干旱胁迫下橡胶树叶片差异表达蛋白的鉴定与功能解析[J].热带作物学报,2009,30(12):1782-1788.Lin X Q,Yuan K,Wang Z H,et al.Identification and Functional Analysis of DIfferentially Expressed Proteins in Hevea brasiliensis Leaves during Drought Stress[J].Chinese Journal of Tropical Crops,2009,30(12):1782-1788.
8 .Wang L F.Physiological and molecular responses to drought stress in rubber tree(Hevea brasiliensis Muell.Arg.)[J].Plant Physiol and Biochemistry,2014,83(2):243-249.
9 .Xue T T,Wang D,Zhang S Z,et al.Genome-wide and expression analysis of protein phosphatase 2C in rice and arabidopsis[J].BMC Genomics,2008,9(1):550.
10 .Landi S,Nurcato R,De Lillo A,et al.Glucose-6-phosphate dehydrogenase plays a central role in the response of tomato(Solanum lycopersicum)plants to short and long-term drought[J].Plant Physiology and Biochemistry,2016,105:79-89.
11 .Zhang F Y,Fu X Q,LüZ Y,et al.Type 2C phosphatase 1of Artemisia annua L.is a negative regulator of ABA signaling[J].Bio Med Research International,2014,2014:521794.
12 .Wei K F,Pan S.Maize protein phosphatase gene family:identification and molecular characterization[J].BMC Genomics,2014,15(1):773.
13 .Stern A,Privman E,Rasis M,et al.Evolution of the metazoan protein phosphatase 2C superfamily[J].Journal of Molecular Evolution,2007,64(1):61-70.
14 .Xing Y N,Xu Y H,Chen Y,et al.Structure of protein phosphatase 2A core enzyme bound to tumor-inducing toxins[J].Cell,2006,127(2):341-353.
15 .Fuchs S,Grill E,Meskiene I,et al.Type 2C protein phosphatases in plants[J].FEBS Journal,2013,280(2):681-693.
16 .Cao J M,Jiang M,Li P,et al.Genome-wide identification and evolutionary analyses of the PP2C gene family with their expression profiling in response to multiple stresses in Brachypodium distachyon[J].BMC Genomics,2016,17(1):175.
17 .阮海华,徐朗莱.2C类蛋白磷酸酶的结构与功能研究进展[J].南京农业大学学报,2007,30(1):136-141.Ruan H H,Xu L L.Progress in the structure and function of PP2C type protein phosphatases[J].Journal of Nanjing Agricultural University,2007,30(1):136-141.
18 .胡晓丽,李德全.植物蛋白磷酸酶2C(PP2C)及其在信号转导中的作用[J].植物生理学通讯,2007,43(3):407-412.Hu X L,Li D Q.Protein Phosphatase 2C in Plants and Its Functions of Signal Transduction[J].Plant Physiology Communications,2007,43(3):407-412.
19 .Chen J H,Zhang D Z,Zhang C,et al.A putative PP2C-encoding gene negatively regulates ABA Signaling in Populus euphratica[J].PLo S One,2015,10(10):e0139466.
20 .Cui M H,Yoo K S,Hyoung S,et al.An Arabidopsis R2R3-MYB transcription factor,At MYB20,negatively regulates type 2C serine/threonine protein phosphatases to enhance salt tolerance[J].FEBS Letters,2013,587(12):1773-1778.
21 .Leung J,Merlot S,Giraudat J.The Arabidopsis ABSCISIC ACID-INSENSITIVE2(ABI2)and ABI1genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction[J].The Plant Cell,1997,9(5):759-771.
22 .Gosti F,Beaudoin N,Serizet C,et al.ABI1 protein phosphatase 2C is a negative regulator of abscisic acid signaling[J].The Plant Cell,1999,11(10):1897-1909.
23 .Schweighofer A,Kazanaviciute V,Scheikl E,et al.The PP2C-type phosphatase AP2C1,which negatively regulates MPK4 and MPK6,modulates innate immunity,jasmonic acid,and ethylene levels in Arabidopsis[J].The Plant Cell,2007,19(7):2213-2224.
24 .Meskiene I,Baudouin E,Schweighofer A,et al.Stress-induced protein phosphatase 2C is a negative regulator of a mitogen-activated protein kinase[J].Journal of Biological Chemistry,2003,278(21):18945-18952.
25 .郭鹏,张士刚,邢鑫,等.欧美杨Pd PP2C基因的克隆与功能分析[J].北京林业大学学报,2015,37(2):100-106.Guo P,Zhang S G,Xing X,et al.Cloning and Function analysis of Pd PP2C from Popululus deltoises[J].Journal of Beijing Forestry University,2015,37(2):100-106.
26 .闵东红,薛飞洋,马亚男,等.谷子PP2C基因家族的特性[J].作物学报,2013,39(12):2135-2144.Min D H,Xue F Y,Ma Y N,et al.Characteristics of PP2C Gene Family in Foxtail Millet(Setaria italica)[J].Acta Agronomica Sinica,2013,39(12):2135-2144.
27 .Vurukonda S S K P,Vardharajula S,Shrivastava M,et al.Enhancement of drought stress tolerance in crops by plant growth promoting rhizobacteria[J].Microbiological Research,2016,184:13-24.
28 .Basu S,Ramegowda V,Kumar A,et al.Plant adaptation to drought stress[J].F1000Research,2016,5:1554.
29 .Chaves M M,Pereira J S,Maroco J,et al.How plants cope with water stress in the field?Photosynthesis and growth[J].Annals of Botany,2002,89(7):907-916.
30 .李国尧,王权宝,李玉英,等.橡胶树产胶量影响因素[J].生态学杂志,2014,33(2):510-517.Li G Y,Wang Q B,Li Y Y,et al.A Review of Influencing Factors on Latex,Yeild of Hevea brasiliensis[J].Chinese Journal of Ecology,2014,33(2):510-517.
31 .Xu Z Z,Zhou G S,Shimizu H.Plant responses to drought and rewatering[J].Plant Signaling&Behavior,2010,5(6):649-654.
32 .Rodriguez P L.Protein phosphatase 2C(PP2C)function in high plants[J].Plant Molecular Biology,1998,38(6):919-927.
33 .Moorhead G B G,Trinkle-mulcahy L,Ulke-lemée A.Emerging roles of nuclear protein phosphatases[J].Nature Reviews Molecular Cell Biology,2007,8(3):234-244.
34 .Bhalothia P,Sangwan C,Alok A,et al.PP2C-like promoter and its deletion variants are induced by ABA but not by Me JA and SA in Arabidopsis thaliana[J].Frontiers in Plant Science,2016,7:547.
35 .Thomashow M F.Plant cold acclimation:freezing tolerance genes and regulatory mechanisms[J].Annual Review of Plant Physiology and Plant Molecular Biology,1999,50:571-599.
36 .Thtiharju S,Palva T.Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana[J].Plant Journal for Cell&Molecular Biology,2001,26(4):461-470.
37 .何亮,李富华,沙莉娜,等.玉米2C型丝氨酸/苏氨酸蛋白磷酸酶(PP2C)活性与耐旱性的关系[J].作物学报,2008,34(5):899-903.He L,Li F H,Sha Li Na,et al.Activity of Serine/Threonine Protein Phosphatase Type-2C(PP2C)and Its Relationships to Drought Tolerance in Maize[J].Acta Agronomica Sinica,2008,34(5):899-903.
38 .Miyazaki S,Koga R,Bohnert H J,et al.Tissue-and environmental response-specific expression of 10 PP2C transcripts in Mesembryanthemum crystallinum[J].Molecular and General Genetics MGG,1999,261(2):307-316.