Monitoring of Gene Expression Profiles and Isolation of Candidate Genes Involved in Pollination and Fertilization in Rice (Oryza Sativa L.) with a 10K cDNA Microarray
- 271 Downloads
To monitor gene expression profiles during pollination and fertilization in rice at a genome scale, we generated 73 424 high-quality expressed sequence tags (ESTs) derived from the green/etiolated shoot and pistil (0–5 h after pollination, 5hP) of rice, which were subsequently used to construct a cDNA microarray containing ca. 10 000 unique rice genes. This microarray was used to analyze gene expression in pistil unpollinated (UP), 5hP and 5DAP(5 days after pollination), anther, shoot, root, 10-day-old embryo (10EM) and 10-day-old endosperm (10EN). Clustering analysis revealed that the anther has a gene-expression profile more similar to root than to pistil and most pistil-preferentially expressed genes respond to pollination and/or fertilization. There are 253 ESTs exhibiting differential expression (e±2-fold changes) during pollination and fertilization, and about 70% of them can be assigned a putative function. We also recovered 20 genes similar to pollination-related and/or fertility-related genes previously identified as well as genes that were not implicated previously. Microarray and real-time PCR analyses showed that the array sensitivity was estimated at 1–5 copies of mRNA per cell, and the differentially expressed genes showed a high correlation between the two methods. Our results indicated that this cDNA microarray constructed here is reliable and can be used for monitoring gene expression profiles in rice. In addition, the genes that differentially expressed during pollination represent candidate genes for dissecting molecular mechanism of this important biological process in rice.
Unable to display preview. Download preview PDF.
- Broekaert, W.F., Cammue, B., De Bolle, M., Thevissen, K. and De Samblanx, G. 1997. Antimicrobial peptides from plants. Crit. Rev. Plant Sci. 16: 297–323.Google Scholar
- Chen, W., Tang, D., Suo, J., Zhang, Y. and Xue, Y. 2001. Expressional profiling of genes related to pollination and fertilization in rice. C.R. Acad. Sci. Paris, Sci. Vie 324: 1111–1116.Google Scholar
- Jansen, M.A.K., Sessa, G., Malkin, S. and Fluhr, R. 1992. PEPC-mediated carbon fixation in transmitting tract cells reflects style-pollen tube interactions Plant J. 2: 507–515.Google Scholar
- Oono, Y., Seki, M., Nanjo, T., Narusaka, M., Fujita, M., Satoh, R., Satou, M., Sakurai, T., Ishida, J., Akiyama, K., Iida, K., Maruyama, K., Satoh, S., Yamaguchi-Shinozaki, K. and Shinozaki, K. 2003. Monitoring expression profiles of Arabidopsis gene expression during rehydration process after dehydration using ca. 7000 full-length cDNA microarray. Plant J. 34: 868–887.CrossRefPubMedGoogle Scholar