Pollen-Specific Expression of Oryza sativa Indica Pollen Allergen Gene (OSIPA) Promoter in Rice and Arabidopsis Transgenic Systems
- 668 Downloads
Earlier, a pollen-specific Oryza sativa indica pollen allergen gene (OSIPA), coding for expansins/pollen allergens, was isolated from rice, and its promoter—upon expression in tobacco and Arabidopsis—was found active during the late stages of pollen development. In this investigation, to analyze the effects of different putative regulatory motifs of OSIPA promoter, a series of 5′ deletions were fused to β-glucuronidase gene (GUS) which were stably introduced into rice and Arabidopsis. Histochemical GUS analysis of the transgenic plants revealed that a 1631 bp promoter fragment mediates maximum GUS expression at different stages of anther/pollen development. Promoter deletions to −1272, −966, −617, and −199 bp did not change the expression profile of the pollen specificity. However, the activity of promoter was reduced as the length of promoter decreased. The region between −1567 and −199 bp was found adequate to confer pollen-specific expression in both rice and Arabidopsis systems. An approximate 4-fold increase in the GUS activity was observed in the pollen of rice when compared to that of Arabidopsis. As such, the OSIPA promoter seems promising for generation of stable male-sterile lines required for the production of hybrids in rice and other crop plants.
KeywordsArabidopsis Cis-regulatory element GUS Pollen-specific promoter Rice Transgenics
This work was supported by a grant from Department of Biotechnology, Government of India. RK and SVK are recipients of JRF and SRF given by Council of Scientific & Industrial Research. We would like to thank Drs. Sanjay Kapoor and Vikrant Gupta for their critical inputs.
- 15.van Tunen, A. J., Koes, R. E., Spelt, C. E., van der Krol, A. R., Stuitje, A. R., & Mol, J. N. (1988). Cloning of the two chalcone flavanone isomerase genes from Petunia hybrida: Co-ordinate, light-regulated and differential expression of flavonoid genes. The EMBO Journal, 7, 1257–1263.Google Scholar
- 23.Twell, D., Wing, R., Yamaguchi, J., & McCormick, S. (1989). Isolation and expression of an anther-specific gene from tomato. Molecular Genetics and Genomics, 217, 240–245.Google Scholar
- 33.Lonsdale, D. M., Allen, R. L., Belostotsky, D., et al. (1995). An analysis of the relative activities of a number of promoter constructs from genes which are expressed during late pollen development as determined by particle bombardment. Plant Cell Reports, 15, 154–158.Google Scholar
- 44.Shcherban, T.Y., Shi, J., Durachko, D.M., et al. (1995). Molecular cloning and sequencing analysis of expansins—a highly conserved, multigene family of proteins that mediate cell wall extension in plants. Proceedings of the National Academy of Sciences USA, 92, 9245–9249.Google Scholar
- 52.Bailey, N. T. J. (1995). Statistical methods in biology (3rd ed.). Cambridge, UK: Cambridge University Press.Google Scholar
- 53.Jefferson, R. A., Kavanagh, T. A., & Bevan, M. W. (1987). GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO Journal, 6, 3901–3907.Google Scholar
- 67.Colot, V., Robert, L. S., Kavanagh, T. A., Bevan, M. W., & Thompson, R. D. (1987). Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. The EMBO Journal, 6, 3559–3564.Google Scholar
- 69.Mitsuhara, I., Ugaki, M., Hirochika, H., et al. (1996). Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants. Plant Cell Physiology, 37, 49–59.Google Scholar
- 70.Albani, D., Sardana, R., Robert, L. S., Altosaar, I., Arnison, P. G., & Fabijanski, S. F. (1992). A Brassica napus gene family which shows sequence similarity to ascorbate oxidase is expressed in developing pollen. Molecular characterization and analysis of promoter activity in transgenic tobacco plants. Plant Journal, 2, 331–342.Google Scholar
- 72.Anand, S., & Tyagi, A. K. (2010). Characterization of a pollen-preferential gene OSIAGP from rice (Oryza sativa L. subspecies indica) coding for an arabinogalactan protein homologue, and analysis of its promoter activity during pollen development and pollen tube growth. Transgenic Research, 19, 385–397.CrossRefGoogle Scholar
- 74.Twell, D., Yamaguchi, J., & McCormick, S. (1990). Pollen-specific gene expression in transgenic plants: Coordinate regulation of two different tomato gene promoters during microsporogenesis. Development, 109, 705–713.Google Scholar