杨树种子萌发中内肽酶变化及其相关途径基因的表达模式
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摘要
为研究杨树种子萌发过程中储藏蛋白分解的分子机制,以杨树萌发0h、0.75h、24h、48h、72h和144h的种子或幼苗为材料,对内肽酶酶活及其相关途径基因的表达模式进行研究,结果表明:随着杨树种子萌发的进行,游离氨基酸含量逐渐升高,其中在48h时变化显著,此时内肽酶活性最高;天冬氨酸内肽酶基因(Potri.006G087600)与苏氨酸内肽酶基因(Potri.002G166800)在48h时相对表达量最高,其中苏氨酸内肽酶基因(Potri.002G166800)表达模式与氨基酸含量变化呈显著正相关,其编码的酶对杨树种子萌发过程中蛋白分解起重要作用。
The endopeptidase activity and expression pattern of related pathway genes in poplar seeds or seedlings germinated at 0 h,0.75 h,24 h,48 h,72 hand 144 hwere analyzed to study the molecular mechanism of storage protein decomposition during germination process of poplar seeds.Result:The free amino acid content gradually increases with extension of germination time and the endopeptidase activity reaches the highest at 48 h.The relative expression of an aspartate endopeptidase gene(Potri.006 G087600)and a threonine endopeptidase gene(Potri.002 G166800 is the highest at 48 h.There is a significant positive correlation between expression pattern of threonine endopeptidase gene(Potri.002 G166800)and variation of amino acid content at 48 hand its coded enzyme plays an important role in protein decomposition during the germination process of poplar seeds.
The endopeptidase activity and expression pattern of related pathway genes in poplar seeds or seedlings germinated at 0 h,0.75 h,24 h,48 h,72 hand 144 hwere analyzed to study the molecular mechanism of storage protein decomposition during germination process of poplar seeds.Result:The free amino acid content gradually increases with extension of germination time and the endopeptidase activity reaches the highest at 48 h.The relative expression of an aspartate endopeptidase gene(Potri.006 G087600)and a threonine endopeptidase gene(Potri.002 G166800 is the highest at 48 h.There is a significant positive correlation between expression pattern of threonine endopeptidase gene(Potri.002 G166800)and variation of amino acid content at 48 hand its coded enzyme plays an important role in protein decomposition during the germination process of poplar seeds.
引文
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[11]曲春浦,刘关君.小黑杨种子中mRNA的鉴定及生物信息学分析[J].贵州农业科学,2017,45(10):12-18.
[12] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J].Methods,2001,25(4):402-408.
[13] MOROHASHI Y,SHIMOKORIYAMA M.Physiological Studies on Germination of Phaseolus mungo Seeds II.Glucose and Organic-acid Metabolisms in the Early Phases of Germination[J].J Exp Bot.,1972,23(1):54-61.
[14] CARRASCO P,CARBONELL J.Changes in the level of peptidase activities in pea ovaries during senescence and fruit set induced by gibberellic acid[J].Plant Physiology,1990,92(4):1070-1074.
[15]丛日晨,赵喜亭,刘晓辉,等.月季切花采后花瓣内肽酶活性的变化[J].园艺学报,2003,30(2):232-235.
[16]胡晋.种子生物学[M].北京:高等教育出版社,2006:1-2.
[17]宋松泉,程红焱,姜孝成.种子生物学[M].北京:科学出版社,2008:168-199.
[18] RAJJOU L,DUVAL M,GALLARDO K.Seed germination and vigor[J].Annu rev plant biol.,2012,63:507-533.
[19] HOWELL K A,NARSAI R,CARROLL A.Mapping metabolic and transcript temporal switches during germination in rice highlights specific transcription factors and the role of RNA instability in the germination process[J].Plant physiol,2009,149(2):961-980.
[2] BEWLEY J D.Seed germination and dormancy[J].Plant Cell,1997,9(7):1055-1066.
[3] BOVE J,JULLIEN M,GRAPPIN P.Functional genomics in the study of seed germination[J].Genome Biol,2001,3(1):10021-10025.
[4] MITSUHASHI W,OAKS A.Development of endopeptidase activities in maize(Zea mays L.)endosperms[J].Plant Physiol,1994,104(2):401-407.
[5] MIFLIN B J,WALLSGROVE R M,LEA P J.Glutamine metabolism in higher plants[J].Curr Top Cell Regul,1981(20):1.
[6]张勇,张守攻,齐力旺,等.杨树——林木基因组学研究的模式物种[J].植物学通报,2006,23(3):286-293.
[7] ECKENWALDER J E.Systematics and evolution of Populus[J].Biology of Populus and its implications for management and conservation,1996(7):32-36.
[8]张帅,谯四红,樊孝萍,等.不同环境条件对黑杨派杨树种子萌发及成苗率的影响[J].安徽林业科技,2014,40(3):15-17.
[9] ZHANG H,WANG W Q,LIU S J,et al.Proteome Analysis of Poplar Seed Vigor[J].Plos One,2015,10(7):e0132509.
[10] ZHANG H,ZHOU K X,WANG W Q,et al.Proteome analysis reveals an energy-dependent central process for Populus×canadensis,seed germination[J].Journal of Plant Physiology,2017,213(6):134-147.
[11]曲春浦,刘关君.小黑杨种子中mRNA的鉴定及生物信息学分析[J].贵州农业科学,2017,45(10):12-18.
[12] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J].Methods,2001,25(4):402-408.
[13] MOROHASHI Y,SHIMOKORIYAMA M.Physiological Studies on Germination of Phaseolus mungo Seeds II.Glucose and Organic-acid Metabolisms in the Early Phases of Germination[J].J Exp Bot.,1972,23(1):54-61.
[14] CARRASCO P,CARBONELL J.Changes in the level of peptidase activities in pea ovaries during senescence and fruit set induced by gibberellic acid[J].Plant Physiology,1990,92(4):1070-1074.
[15]丛日晨,赵喜亭,刘晓辉,等.月季切花采后花瓣内肽酶活性的变化[J].园艺学报,2003,30(2):232-235.
[16]胡晋.种子生物学[M].北京:高等教育出版社,2006:1-2.
[17]宋松泉,程红焱,姜孝成.种子生物学[M].北京:科学出版社,2008:168-199.
[18] RAJJOU L,DUVAL M,GALLARDO K.Seed germination and vigor[J].Annu rev plant biol.,2012,63:507-533.
[19] HOWELL K A,NARSAI R,CARROLL A.Mapping metabolic and transcript temporal switches during germination in rice highlights specific transcription factors and the role of RNA instability in the germination process[J].Plant physiol,2009,149(2):961-980.