Analysis Of Grouting Characteristics Transcription And Proteomics In TaGW2-6A Coding Region Allelic Variation Lines

In recent years, the grian wright gene TaGW2-6A has been widely studied. Previous reports showed a single‘T’base insertion in the eighth exon of the grain weight gene TaGW2-6A cause increasing in the grain width and weight. Base on this result, we analysis the grouting characteristics, transcriptional pattern and proteomic difference during seven grain development stages( i.e., at 3 Day after anthesis(DAA), 6DAA, 9DAA, 12 DAA, 15 DAA, 20 DAA, 25DAA)in TaGW2-6A coding region allelic variants though Chinese spring and NIL-31..The main results are concluded as follows:1. The grain size and weight increased gradually from 3 to 25 DAA in both CS and NIL-31. Either KL or TKW differed significantly(P ? 0.01) between CS and NIL-31 in all seven grain development stages. Excluding 3 DAA, KW differed significantly between CS and NIL-31 in the other six stages(P ? 0.01). Meanwhile, the NIL-31 shows a higher biomass accumulation rate than CS from 3 DAA to 15 DAA, especially from 6 DAA to 12 DAA. However, the biomass accumulation rate shows nearly equally between CS and NIL-31 from 15 DAA to 25 DAA.These results further confirmed that the insertion mutation was a super allelic variant and improve the biomass accumulation rate in the early development period.2.The transcript abundance levels of the TaGW2-6A coding region allelic variants differed significantly by several orders of magnitude, which showed that Ta GW2-6A is negatively associated with grain width. During the seven stages of grain development. Each allelic variants of TaGW2-6A reached their first transcription peak at 6 days after anthesis(DAA), which was the start of the endosperm cell division time point. This suggested that TaGW2-6A had an essential role in regulating cell differentiation. However, the insertion type of TaGW2-6A allelic reached the second peak was earlier than the normal type, i.e., at 12 DAA and 20 DAA, respectively. The grain filling stage usually from 12 DAA and 21 DAA. Therefore, the intertion mutation allelic variant may have a role of coordinated regulation of endosperm cell d e v e l o p m e n t a n d g r a i n f i l l i n g w h e r e c o u l d e n h a n c e g r a i n w e i g h t.3. In total, we identified 228 differentially accumulated protein unique proteins by twodimensional gel electrophoresis, of which, 48 spots were up-regulated in NIL-31, 67 spots were down-regulated in NIL-31, 63 spots were unique in NIL-31 and that of 50 spots in Chinese spring. The proteins detected belonged to 14 main groups via tandem MALDI-TOF/TOF-MA and biological information processing, i.e., carbohydrate metabolism, amino acid biosynthesis, cell development, energy production and transportation, fatty acid biosynthesis, ubiquitin-dependent protein catabolic process, hormone regulation, photosynthesis, protein synthesis/assembly/ degradation, signal transduction, storage protein, stress/defense, transcription/translation, and other unknown functional groups.4. The 26 S proteasome relieve the suppression of DELLA protein in gibberellin signal transduction and promote GA transduction in plant. In this experiment, we found that four alpha proteasome proteins, this enzyme play structural roles in the maintenance of the 20 S and 26 S. At the same time, we found a NIL-31 high abundance of gibberellic acid receptor protein expression GID1L2 protein(4211), possibly due to the efficient DELLA protein degradation by 26 S proteasome. Therefore, the mutant TaGW2-6A protein might improve the level of GA signal transduction via the regulation of 26 S.5. We find that TaGW2-6A protein may enhance stress tolerance via induce various types of mutations. Function analysis shows that the proteasome plays an important role in the process of cell differentiation. This suggests that in the process of seed growth Ta GW2-6A mutant protein can through induction of cell differentiation to enhance the plants stress tolerance.