Conformation Of Genes Expressing Specifically In Rice Endosperm And Their Application

In the study, rice endosperm library, its bioinformatics database and 966 different rice endosperm total RNA from Jiaxing Academy of Agricultural Sciences and Institute of Biotechnology and Nuclear Technology, Sichuan Academy of Agricultural Sciences, were used.576 TUTs were annotated after ESTs searching against oryza sativa database of NCBI. Of them, most highly-expressed genes encoded storage proteins, starch, ribosome proteins, initiation factors, elongation factors and transcription factors.In order to analyze genes expression profiles of different rice tissues and to find the highly-expressed genes in endosperm, 2013 TUTs were printed on cDNA array. And different tissues included endosperm, embryo, ear, leaf and stem internode during anthesis and 10-15 days after anthesis. Differential expression analysis, especially comparing endosperm with other tissues, was performed by Cyber-T Bayesian statistical framework. While Bayesian p-value was set to be 0.05, expression of 203 TUTs changed remarkablly. Except for 129 down-regulated genes, a total of 101 TUTs were up-regulated prominently in endosperm, and 75 TUTs were annotated. The up-regulated genes were mostly concerned with storage protein and starch synthesis. There were 29 TUTs encoding storage protein, such as glutelin and prolamin. 4 TUTs participated in amylose and amylopectin synthesis. Other genes mostly encoded ct-amylase, light-inducible protein, transduction factor, signal recognition particle and so on. Unkown genes occupied about a quarter of all up-regulated genes.And the veracity of cDNA array was validated by following Northern blot hybridization. The result had confirmed the credibility of cDNA-array. And glutelin, prolamin, lipid transfer protein, soluble starch synthase, 18 kDa oleosin and ot-amylase were found only expressed in endosperm, the same as the larger Rbel transcription.The hybridization with RNA array of 966 RNA indicated that Wx gene had linear relationship with rice AC on the whole. But some rice samples were not coincident with the rule. Wx gene mature mRNA and an un-mature mRNA were tested by Cyber-T. And the result indicated that fold had positive relationship with AC. AC was 17%-20% while fold between -1 and +1.19 genes expression profiles were analyzed by RNA array. These genes coded storage protein, starch and lipid. And we found the expression of homologous genes may be different.According to the expression profile of four glutelin genes, the expression of H105D02 is similar to GluB, and the expression of H063F03 simialr to GluA.Two unkown genes (H098F12, H051G06) and other 10 known genes were clustered. And the result indicated that H098F12 had closer expressional pattern with allergenic protein (H097G05).H051G06 was similar to 10 kDa oleosin (H129F01), soluble starch synthase (H144A06), and 13kDa prolamin (H110F01). We deduced they were respectively co-regulated genes. And northern had confirmed H051G06 as endosperm-specific gene; we deduced it was concerned with storage substance genes.Our results could provide reference for rice breeding, because a large number of samples were generations during breeding. For example, ZRF6 was one of F1 of early rice from Jiaxing agricultural academy of sciences. The expression of prolamin, glutelin and the genes concerned with starch synthesis in ZRF6 was higher than other samples of Fl of early rice. This could direct to select high-quality F2. And this was also adapt to new rice combination of 2003 and fertility restoring line select of 2003 from Sichuan agricultural academy of sciences.