草莓果实中FaPP2CX1基因克隆与表达分析
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摘要
【目的】为了探究FaPP2CX1在非呼吸跃变型草莓果实成熟中的作用。【方法】以八倍体草莓‘甜查理’为材料,通过RT-PCR克隆FaPP2CX1编码序列,生物信息学工具分析序列特征,其次通过qRT-PCR分析不同时期草莓果实(大绿期、白果期、始红期、片红期和全红期)中FaPP2CX1基因的表达量。【结果】FaPP2CX1基因全长为3 765bp,编码1 254个氨基酸,有丝氨酸苏氨酸激酶的N-末端(STK_N)结构域、富亮氨酸重复蛋白(LRR)结构域、核苷酸结合(NB-ARC)结构域和蛋白磷酸酶(PP2C)结构域四个保守结构域,是草莓中独有的基因。从大绿期开始,FaPP2CX1的表达量逐渐降低,始红期到达最低值;而后随着草莓果实着色加深,表达量迅速上升,并在果实成熟(即全红期)达到最高。【结论】暗示FaPP2CX1可能参与草莓果实的成熟调控。
【Objective】To investigate the role of protein phosphatase 2 Ctranscript variant X1(FaPP2CX1)in the ripening of non-climacteric strawberry fruit.【Method】In this study,the octoploid strawberry'Sweet Charlie'(Fragaria×ananassa)was used as the material.Firstly,the FaPP2CX1 coding sequence was cloned by RT-PCR,and bioinformatics methods were used to analyze the sequence characteristics.Secondly,the expression levels of FaPP2CX1 were analyzed in different periods(large green,white,initial red,partial red,and full red)by qRT-PCR.【Results】The results showed that the FaPP2 CX1 gene has3 765 bp in length and encodes 1 254 amino acids.It has four conserved domains including the N-terminal domain of serine threonine kinase(STK_N),the leucine-rich repeat protein(LRR)domain,a nucleotide binding(NB-ARC)domain,and protein phosphatase(PP2C)domains,respectively,and it is a unique gene in strawberry(Fragaria).From the beginning of the large green period,the expression level of FaPP2 CX1 gradually decreased,and reached the lowest point at the initial red period,then increased rapidly in consistent with red-coloring,and reached the highest level in ripened fruits(full red period).【Conclusion】Our results suggest that FaPP2 CX1 may be involved in the regulation of strawberry fruit ripening.
【Objective】To investigate the role of protein phosphatase 2 Ctranscript variant X1(FaPP2CX1)in the ripening of non-climacteric strawberry fruit.【Method】In this study,the octoploid strawberry'Sweet Charlie'(Fragaria×ananassa)was used as the material.Firstly,the FaPP2CX1 coding sequence was cloned by RT-PCR,and bioinformatics methods were used to analyze the sequence characteristics.Secondly,the expression levels of FaPP2CX1 were analyzed in different periods(large green,white,initial red,partial red,and full red)by qRT-PCR.【Results】The results showed that the FaPP2 CX1 gene has3 765 bp in length and encodes 1 254 amino acids.It has four conserved domains including the N-terminal domain of serine threonine kinase(STK_N),the leucine-rich repeat protein(LRR)domain,a nucleotide binding(NB-ARC)domain,and protein phosphatase(PP2C)domains,respectively,and it is a unique gene in strawberry(Fragaria).From the beginning of the large green period,the expression level of FaPP2 CX1 gradually decreased,and reached the lowest point at the initial red period,then increased rapidly in consistent with red-coloring,and reached the highest level in ripened fruits(full red period).【Conclusion】Our results suggest that FaPP2 CX1 may be involved in the regulation of strawberry fruit ripening.
引文
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[13]张雪薇,刘仑,鲁黎明,李立芹.烟草磷酸酶基因NtPP2C16的克隆、表达载体构建及表达分析[J].华北农学报,2017,32(5):82-89
[14]郭鹏,张士刚,邢鑫,姜健.欧美杨PdPP2C基因的克隆与功能分析[J].北京林业大学学报,2015,37(2):100-106
[15]Wang QH,Zhao C,Zhang M,Li YZ,Shen YY,Guo JX.Transcriptome analysis around the onset of strawberry fruit ripening uncovers an important role of oxidative phosphorylation in ripening[J].Scientific Reports,2017,7:41477
[16]Wang GL,Wu C,Zeng L,He C,Baraoidan M,de Assis Goes da Silva F,Williams CE,Ronald PC,Leung H.Isolation and characterization of rice mutants compromised in Xa21-mediated resistance to X.oryzae pv.oryzae[J].Theoretical Applied Genetics,2004,108(3):379-384
[17]Hou BZ,Xu C,Shen YY.A leu-rich repeat receptor-like protein kinase,FaRIPK1,interacts with ABA receptor,FaABAR,to regulate strawberry(Fragaria×ananassa)fruit ripening[J].Journal of Experimental Botany,2018,69(7):1569-1582
[18]He Z,Wang ZY,Li J,Zhu Q,Lamb C,Ronald P,Chory J.Perception of brassinosteroids by the extracellular domain of the receptor kinase BRI1[J].Science,2000,288(5475):2360-2363
[19]Wang X,Li X,Meisenhelder J,Hunter T,Yoshida S,Asami T,Chory J.Autoregulation and Homodimerization Are Involved in the Activation of the Plant Steroid Receptor BRI1[J].Developmental Cell,2005,8(6):855-865
[2]朱海生,李永平,温庆放,潘东明,林义章.草莓遗传转化研究进展[J].基因组学与应用生物学,2009,28(2):408-416
[3]刘建成,段可,李静,叶正文,苏建宇,高清华.草莓NBS-LRR家族基因FaNBS1的克隆与表达分析[J].果树学报,2011,28(6):1025-1031
[4]祝军,柴璐,侯柄竹,沈元月,郭家选.果实中脱落酸的研究进展与展望[J].园艺学报,2015,42(9):1664-1672
[5]贾海锋,赵密珍,王庆莲,房经贵,赵鹏程,刘众杰,张成,纠松涛.生长素和脱落酸在草莓果实发育过程中的作用[J].江苏农业科学,2016,44(11):173-176
[6]Chai YM,Jia HF,Li CL,Dong QH,Shen YY.FaPYR1is involved in strawberry fruit ripening[J].Journal of Experimental Botany,2011,62(14):5079-5089
[7]Jia HF,Lu D,Sun JH,Li CL,Xing Y,Qin L,Shen YY.Type 2Cprotein phosphatase ABI1is a negative regulator for strawberry fruit ripening[J].Journal of Experimental Botany,2013,64:1677-1687
[8]Li CL,Jia HF,Chai YM,Shen YY.Abscisic acid perception and signaling transduction in strawberry:A model for nonclimacteric fruit ripening[J].Plant Signaling and Behavior,2011,6(12):1950-1953
[9]姜丽,冷平.关于ABA信号转导核心PP2C的研究进展分析[J].中国农业文摘-农业工程,2017,(5):17-25
[10]Nakashima K,Yamaguchi-Shinozaki K.ABA signaling in stress-response and seed development[J].Plant Cell Reports,2013,32(7):959-970
[11]Liu L,Hu X,Song J,Zong X,Li D,Li D.Overexpression of a Zea mays L.protein phosphatase 2Cgene(ZmPP2C)in Arabidopsis thaliana decreases tolerance to salt and drought[J].Journal of Plant Physiology,2009,166(5):531-542
[12]胡伟,颜彦,何艳臻.小麦TaPP2C59基因的克隆及表达分析[J].麦类作物学报,2014,34(10):1334-1340
[13]张雪薇,刘仑,鲁黎明,李立芹.烟草磷酸酶基因NtPP2C16的克隆、表达载体构建及表达分析[J].华北农学报,2017,32(5):82-89
[14]郭鹏,张士刚,邢鑫,姜健.欧美杨PdPP2C基因的克隆与功能分析[J].北京林业大学学报,2015,37(2):100-106
[15]Wang QH,Zhao C,Zhang M,Li YZ,Shen YY,Guo JX.Transcriptome analysis around the onset of strawberry fruit ripening uncovers an important role of oxidative phosphorylation in ripening[J].Scientific Reports,2017,7:41477
[16]Wang GL,Wu C,Zeng L,He C,Baraoidan M,de Assis Goes da Silva F,Williams CE,Ronald PC,Leung H.Isolation and characterization of rice mutants compromised in Xa21-mediated resistance to X.oryzae pv.oryzae[J].Theoretical Applied Genetics,2004,108(3):379-384
[17]Hou BZ,Xu C,Shen YY.A leu-rich repeat receptor-like protein kinase,FaRIPK1,interacts with ABA receptor,FaABAR,to regulate strawberry(Fragaria×ananassa)fruit ripening[J].Journal of Experimental Botany,2018,69(7):1569-1582
[18]He Z,Wang ZY,Li J,Zhu Q,Lamb C,Ronald P,Chory J.Perception of brassinosteroids by the extracellular domain of the receptor kinase BRI1[J].Science,2000,288(5475):2360-2363
[19]Wang X,Li X,Meisenhelder J,Hunter T,Yoshida S,Asami T,Chory J.Autoregulation and Homodimerization Are Involved in the Activation of the Plant Steroid Receptor BRI1[J].Developmental Cell,2005,8(6):855-865