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Gene Expression Profiling of Nils Response to Xanthomonas Oryzae Pv. Oryzae and Improvement of Resistance to Bacterial Blight in Rice Restorer Line

Author: LiuBingXin
Tutor: YangChangDeng
School: Chinese Academy of Agricultural Sciences
Course: Crop Genetics and Breeding
Keywords: Rice Bacterial blight Backcross Marker-assisted selection Near-isogenic lines Gene chip Improved varieties
CLC: S511
Type: Master's thesis
Year: 2010
Downloads: 146
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Abstract


Bacterial blight is a bacterial disease of rice production on the most harmful one. The most cost-effective method against rice bacterial blight resistance gene cultivating resistant varieties, a disease-resistant varieties of continuous cultivation of a certain period of time, however, often lose resistance. To make better use of bacterial blight resistance gene to study the resistance mechanisms of bacterial blight resistance gene as well as the utilization of the resistance gene is imperative. Rice bacterial blight resistance mechanism is very complicated. With the completion of the sequencing of the rice genome, gene chip technology matures and rice functional genomics research, using DNA microarray analysis of rice in pathogen stress gene expression profile has become a reality. Xa23 is currently using the most widely used anti broadest spectrum of one of the resistance genes. In order to understand the mechanism of resistance of the resistance gene Xa23 and to provide information to help clone the gene, the study selected carrying Xa23 the BR1 and restorer lines parental Shuhui527 use of backcross, selfing combined with marker-assisted selection methods to establish white leaf blight resistance gene Xa23 anti sense of near-isogenic lines. Using Affymetrix rice microarray (containing 57,381 probes) genome-wide expression profiling analysis of bacterial blight infection 12h after bacterial blight resistant and susceptible NILs expression. The results are as follows:. Distributed 522 pairs of 12 chromosomes SSR primer screening Shuhui527 between BR1 parents, to the polymorphic primers 87 pairs of these polymorphic primers screening candidate groups to two lines ( L16, L17) differences in bacterial blight resistance gene Xa23 locus. Agronomic traits investigated also showed that the two strains were no significant differences Near excellent. 2. Susceptible material L17 control a total of 911 differentially expressed genes in resistant materials L16, of which 753 genes were up-regulated and 158 genes were down-regulated. Up-regulated genes the significant excess downregulated genes. Gene ontology classification results can be seen, the differentially expressed genes involved in cell parts, cells, physiological processes, cellular processes and metabolism. KEGG pathway analysis of differentially expressed gene ubiquinone biosynthesis, ribosomes, oxidative phosphorylation, steroid synthesis, the metabolism of amino acids, benzoic acid metabolic pathway of coenzyme A, cytochrome P450 metabolism, inositol phosphate signaling system. 3. L16 vs L17 753 up-regulated genes, we found 653 gene predictive function, annotated as protein expression, but 14 of unknown function annotated as hypothetical proteins 217, does not have any comments total 100. 158 down-regulated genes, we found that the prediction function 139 Notes for the expression of the protein, but three of unknown function, annotated as hypothetical proteins 30, no annotation information for a total of 19. Two encoding disease resistance gene family proteins (Os11g0588600, Os11g0640300) in L16vsL17 upregulated genes are located on chromosome 11, 7 encoding cytochrome P450 genes, 16 genes encoding proteins type the same as protein kinases, three genes encoding oxidoreductase and 2 encoding Myb transcription factor genes, three encoding thaumatin pathogenesis-related protein gene; downregulated genes the two encoding cytochrome P450 family protein genes, three genes encoding proteins for protein kinases. Intrinsic molecular mechanism to explain rice by bacterial blight infection. In addition, this study will also bacterial blight resistance gene Xa23 backcrossing Yihui 1577 restorer lines after backcross prospects choose from the cross and random selection criteria, in BC1F7 and the BC2F6 in genome-wide background detection and agronomic traits investigated, analyzed BC1F7 and BC2F6 populations genetic background response rate and agronomic traits reply; offspring of the backcross improved performance of the superior part of the material and Sterile the Tianfeng A measured with the investigated combining ability recovery degrees and heterosis breeding advantages combination. The results are as follows: 1. Yihui 1577 between BR1 parents, 522 pairs of SSR primers were screened to 105 pairs of polymorphic primers located on chromosome 12 polymorphic ratio was 20.11%. 105 pairs of SSR polymorphism primers to detect the genetic background of the 13 BC1F7 and 11 BC2F6 lines. The results showed that the BC1F7 13 strains, genetic background response rate of 60.10% -74.29%; distribution with the theory of genetic background response rate in the BC2F6 the 11 lines, the genetic background response rate at 81.43% -91.43% 87.25% or so. That randomly selected BC1F7 and BC2F6 strains of genetic background response rate response rates close to the theoretical genetic background. Bacterial Blight Resistance of BC1F7 and BC2F6 improved restorer lines have been improved, the excellent bacterial blight resistance. A BC1F7 the 13 strains in ear length and leaf length traits on all lines with the recurrent parent Yihui 1577 showed no significant differences in the traits of effective panicles and leaf width in addition to the individual lines the rest are close Yihui 1577, no significant difference. BC2F6 11 lines, all the lines on the traits of effective panicles and flag leaf length, no significant difference; most of the traits of panicle length and seed setting rate was no significant difference. The BC2F6 lines improved offspring upgrading offspring than BC1F7 lines in effective panicles, spikelets per panicle, seed setting rate, grain weight and other closely related traits and yield closer to the recurrent parent. This recovery rate in the BC1F7 is consistent with the genetic background of the BC2F6. 3. BC1F8 testcross species, effective panicles, leaf length and leaf width traits all showed no significant difference; no significant difference in most of the spike length, grain number per plant theoretical yield traits ; seed setting rate and grain weight control significant differences by donor parents a greater impact. A BC2F7 the test cross species effective panicle traits measured breeding of all strains had no significant difference. Most there was no significant difference in spike length, spikelets per panicle, seed setting rate, the theoretical yield per plant, leaf length and leaf width traits. Higher than control, and only LT81, LT83 and LT86 measured mating significant difference in the performance of all grain weight traits. Gained 3 with a yield of bacterial blight resistance and control similar to hybrid rice combination, may be applied to production.

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