Related Proteome Analysis Of Overexpressing OsCYP2Mutant In Response To Salt Stress

Japanese japonica rice varieties Aichi were selected as the material, through transgenic technology,overexpressing OsCYP2gene, salt-tolerance transgenic Aichi Asahi Material:OE, OE and wild-type Asahi as the experimental materials,using the following two biological technology:two dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (M ALDI-TOF-MS/MS), respectively, in the normal and salt stress (200mM NaCl) early seedlings (10d seedling age), the salt stress response proteome analysis and identification of the experimental results are as follows:1.Transgenic rice varieties OE and receptor AZX were planted in germination boxs that containing water (control) and200mM NaCl,respectively.After10days,, taking the above-ground sheet byTCA-acetone precipitation method in order to extract thtotal protein, using two different PH section (3-10and5-8)17cm linear IPG strips to isolate the total protein. The experimental results show that two rice2-DE protein expression spectrum reproducibility are very good. By PDQuest software analysis, the matching rate surpass90%.Fifty-six spots were significantly differential express found in mutant leaves compared with the control.Theywere selectedand identified by MALDI-TOF-MS/MS, Only thirty-five uniproteins,such asribulose-1,5-bisphosphate carboxylase/oxygenase, Ascorbate Peroxidase, abscisic stress ripening, caffeoyl-CoA O-methyltransferase and chlorophyll a-b binding protein, were identified.These proteins are involved in carbohydrate and energy metabolism regulation, disease control, ROS detoxification and proteinsynthesis and localization.2.Considering proteins’function and differential expressing level,9proteins are chosen to characterize from mRNA expressing level:including4protein pots which are up in transgenic mutant and down in wild Asahi(2101,1106,6104and2603),4protein pots which are up significantly in transgenic mutant but not change in wild Asahi(2208,1102,7408and1402) and1protein pot which are down significantly in transgenic mutant and up in wild Asahi(4107).They are involed to many biology process:protein hydrolyzation, electron transportion, amino acid synthesis and carbohydrate metabolism.The results of characterizing from mRNA expressing level show that:3proteins keep in the same absolutely,which change in transcription level and translation level;4of them keep in the same basically;The others are opposite.Above all,Two japonica rice combinations,salt stress responsive proteins,keep in the same basically in transcription level and translation level.The study compares mutant and wild Asahi’proteins differences under normal and salt stress conditions,analyzes Physiological and Biochemical Mechanism of mutant salt tolerance’s increasing and discovers salt-related proteins. It not only can analyze the correlation between differential proteins and OsCYP2gene’s insertation but also benefit for discovering OsCYP2gene’s function and salt-tolerance mechanism.