| commonly used in medicinal materials in our country.Its production,however,is affected chronically by consecutive monoculture,which causes poor plant growth,fibrous root roots,abnormal tuberous root,obvious yield and quality decrease and,moreover with no harvest,and harm period is long,sustainable for 8 to 10 years.Consecutive monoculture problem has become the major obstacle which restricts the development of the Rehmanniae industry in China.Previous studies mainly focus on the change of soil environmental after consecutive monoculture and the differences of gene expression in the plant body,differential protein expression profile in the body of consecutive monoculture Rehmannia,especially differential phosphorylation protein expression profile are rarely reported.This research usingiTRAQ technology build consecutive monoculture Rehmannia differential phosphorylation protein expression profile,analyzed the protein phosphorylation expression differences between monoculture treatment and contract in tuber early expansion period in order to obtain the protein phosphorylated responding to consecutive monoculture problems in root and its signal transduction system.The main content of this study are as follows:1.The identification of differential expression phosphorylated proteins in consecutive monoculture Rehmannia root.Analysis and relative quantification of protein phosphorylation by iTRAQ technique,the identification of protein numberswhich Global FDR Fit < 1% is 1787.According to standards of unused>1.3,the number of peptides≥1,the library data with“R”and empty data,data filtering is carried out to obtain high quality data.A total of 1363 credible protein were acquired.According to p-value < 0.05 and the change ratio of 1.2 times the standard screening of differentially expressed proteins,that is,p-value less than 0.05,the change ratio of more than 1.2 times or less than 0.83 are selected as for differential expression of phosphorylated proteins.And the dynamic analysis(with Dynamics Software)of the same proteins in different groups were conducted,in any one group of proteins for the difference in the protein are listed.A total of 78 PTR_PCR(the first year cropping to the second year cropping)proteins,in which 30 up regulated expression and 48 down regulated expression.2.The functional analysis of differential expression phosphorylated protein in consecutive monoculture Rehmannia root.The phosphorylated protein data were analyzedusing KEGG pathway analysis.The result showed that the differential expression phosphorylated proteins are distributed in the 20 metabolic pathways.The major metabolic pathways were as follows: metabolic pathways,secondary metabolic pathways,the ribosome biosynthesis,plant-pathogeninteraction,endoplasmic,protein processing in endoplasmic Rehmannia glutinosa L is perennial herbaceous plantsof Sreophulariaceae,which is one of reticulum and signal transduction pathways,etc.Of which metabolic pathways contained 6differential expression phosphorylated proteins,there are NADP-ME4,SPS1,dl4665 w,TPS5,UKL1 and PLC2.Only dl4665 w was lower expression,the otherswere raised to expression.Signal transduction pathwaycontained 9differential expression phosphorylated proteins,i.e.EHD2,EPSIN2,AGD12,PEN2,PLC2,AGD9,AGD6,CRT2 and TUBA6,the top five were raised to express,the othersdecreased expression.There were 3differential expression phosphorylated proteins associated protein synthesis in endoplasmic reticulum,i.e.PUX4、CRT2 and At1g19130,and all of them were lower expression.2differential expression phosphorylated proteins in plant pathogen interactions,CPK6 and MKK2,wereup-expression.Interaction analysis of 78 differentially expressed proteins of PTR_PCR protein were carried out,and it were found to thatamong of 34 proteins might happen interaction reaction.Interaction proteins were mainly involved in follow pathways,protein processing in endoplasmic reticulum,phagosome,plant-pathogen interaction,endocytosis,spliceosome,pyruvate andphosphatidylinositol signaling system.3.The possible key signaling pathways forming consecutive monoculture problem in Rehmannia.In specific phosphorylated proteins which response to consecutive monoculture problems,we found that two differential expression phosphorylated proteins,PEN2 and PLC,two key members in phosphatidyl inositol signaling pathways,were up-regulated expressed in response to consecutive monoculture problems.It means the production of messenger IP3,the unlock of the calcium channel,the transfer allelochemicalsfrom cxcellular regionto intracellular andactivation of calcium channel,resulting increase intracellular calcium ion concentration.The result clearly shows that phosphatidyl inositol signal system plays an important role in the forming of Rehmanniaconsecutive monoculture problems.At the same time,four phosphoproteins were also found,ACA9,MKK2,CPK6 and CRT2,which are related to calcium signaling pathways.First three,Located incytoplasmic membrane,were up-expressed significantly and,but CRT2 significantly down-expressed.These phosphorylation proteins may cause the disorder of calcium signal system,the occurrence of aggravated the symptoms of damage,suggesting that Ca signals play an important role in environment signal transmission and transport in formingconsecutive monoculture problems in Rehmannia.L... |
