A Study On Gene Differential Expression Of Camellia Sinensis Before And After Spraying Chemical Pesticides Genes Using CDNA-AFLP

Chemical prevention and control is one of the indispensable means in gardenmanagement of tea plant, when the people benefit,at the same time,people shoule payattention to the negative influence Whether there about the circadian biochemicalreactions and own defense system to disease and insect. This study based on cDNA-AFLP technology, researched the gene expression profile differences of the tea treebefore and after spraying chemical pesticide imidacloprid, and the bioinformaticsanalysis of differentially expressed genes and qRT-PCR verification. Main results asfollows:(1)Using enzymes analysis software, analysis the relevant logined cDNAfull-length sequence and EST of tea in GenBank, results show that the ApoI/MseI isthe best combination using cDNA AFLP technology analysis differentially expressedspectrum on tea tree.(2) By the optimized cDNA-AFLP technology analysis the gene expressionprofile differences of the tea tree before and after spraying chemical pesticideimidacloprid (factory recommend using concentration), results won a total of220bardifferentially expressed frangments (TDF),after the recovery, cloning andsequencing,geting160bar, including the induction expression94, there were66inhibited. The length of differentially expressed frangments amongs113-822bp, mostdistribute between200to300bp and300to400bp, respectively take ua39%and30%, and there were some clips are more than500bp.(3) Using BLAST analysised the homology and found that except8TDF of the160results appear repeat, in the rest152,the functions of26are unclear,18are newno matching; For the rest108,the function prediction results show that these bandsrespectively attributable to the signaling, photosynthesis and energy metabolism,transcription regulation, disease resistance and ars protein, transmembranetransportation, stress reaction and material metabolic7function types. Among them,the induced and inhibited TDF, its belong to function in both types ofphotosynthesis and energy metabolization are the same high, proportion respectivelyare36%and41%; And the difference between them is that transcription regulationfunction related gene of the former obviously less,and the proportion respectively9%and16%; The distinction of the proportion between other function types genes are alittle. (4) In order to verify the correctness and the influencing characteristics ofimidacloprid induced different bands of tea, respectively chose two induced TDF(17N326P5) and four inhibited TDF (12G18J225D1and12O1), analysis by qRT-PCR. Results showed that the expression of the induction TDF raised up tomaximum at the second day, then decreased, at the8th day the difference with thecontrol is not obvious; the inhibited TDF, the expression at the4th day down tothe lowest, then pick up gradually, at the8th day the difference with the control is notobvious.