Studying osm1/ syp61 gene in arabidopsis under drought stress
Keywords:
OSM1, SYP61, T-DNA, drought stress, mutant lineAbstract
Drought is a major abiotic stress that causes a reduction in plant growth and yields of agricultural crops. Naturally, Plants differ in their susceptibility to drought adaptation and tolerance and research confirmed that many genes and transcription factors are involved in the response to water deficit. I tested Arabidopsis OSM1 (for osmotic stress-sensitive mutant 1) (At1g28490) gene- which is similar to SYP61 (SYNTAXIN OF PLANTS 61) -under drought stress for the first time in plants grown on soil. The investigations included wild type plants and osm 1 mutant line (SALK_107167) subjected to drought. The sequence analysis showed that the T-DNA in this mutant is inserted in the promoter and in the correct site as reported by the database but showed smaller product size than the expected confirming missing of 86 bases at the left border of the T-DNA. Genotyping investigations showed that the lines segregated out according to the mendelian bases. The mutant plants were significantly more sensitive to wilting when grown with limited soil moisture compared to wild-type plants but plant survival was not affected. These results indicate that this gene may be involved in drought stress response and wider experimentation is needed to include transcript accumulations under such kind of stress in parallel with the use of a mutant that carries a T-DNA within the gene.
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References
[1.] Alonso J. M. and Stepanova A. N. (2003). T-DNA mutagenesis in Arabidopsis. Methods in molecular biology (Clifton, N.J.) 236 : 177-188.
[2.] Bowen R. (2000). Agarose gel electrophoresis of DNA. Available at: Boyer J. S. (1982) Plant productivity and environment.Science, 218:443-448
[3.] Boyes D.C., Zayed A.M., Ascenzi R., McCaskill A. J., Hoffman N. E., Davis K. R., and Gorlach J. (2001). Growth stage-based phenotypic analysis of Arabidopsis: A model for high throughout functional genomics in plants. Plant Cell 13: 1499-1510.
[4.] Bray E.A. (1997). Plant responses to water deficit. Trends in Plant Science 2: 48-54.
[5.] Chenna R., Sugawara H., Koike T., Lopez R., Gibson T.J., Higgins D. G., and Thompson J.D. (2003). Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res. 31: 3497-3500.
[6.] Edwards K., Johnstone C., and Thompson C. (1991). A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res. 19 : 1349.
[7.] Elhaj A. (2009), Molecular and Physiological investigations of Arabidopsis mutant lines under abiotic stress. School of Biology, Newcastle University, Newcastle upon Tyne, UK. A PhD thesis.
[8.] Harb A., Krishnan A., Ambavaram M. M. R., and Pereira A. (2010). Molecular and physiological analysis of drought stress in Arabidopsis reveals early responses leading to acclimation in plant growth. Plant Physiol. 154: 1254-1271.
[9.] LeshemY., Golani Y., Kaye Y., and Levine A. (2010). Reduced expression of the v-SNAREs AtVAMP71/AtVAMP7C gene family in Arabidopsis reduces drought tolerance by suppression of abscisic acid-dependent stomatal closure.Journal of Experimental Botany. 61 : 2615–2622.
[10.] Leung J.and Giraudat J. (1998). Abscisic acid and signal transduction. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 199-222.
[11.] Krysan P.J., Young J.C., and Sussman M.R. (1999) T-DNA as an insertional mutagen in Arabidopsis. Plant Cell 11: 2283-2290.
[12.] Feldmann K. A. (1991). T-DNA insertion mutagenesis in Arabidopsis: Mutational spectrum. Plant Journal 1: 71-82.
[13.] Gandhi R., Matheshwari S.C. ,Khurana J., and Khurana P. (2001). Genetic and molecular analysis of Arabidopsis thaliana (ecotype ‘Estland’) transformed with Agrobacterium vitro cell. Develop. Biol. Plant .37: 629-637.
[14.] Gheysen G., Villarroel R., and Van Montagu M. (1991). Illegitimate recombination in plants: A model for T-DNA integration. Genes and Development 5: 287-297
[15.] Hall T.A. (2005). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. 41:95-98.
[16.] Higgins D. G. and Sharp P.M. (1988). CLUSTAL: A package for preforming multiple sequence alignment on a microcomputer. Gene 73: 237-244.
[17.] Hobbs S. L. A., Kpodar P., and Delong C.M.O. (1990) The effect of T-DNA copy number, position and methylation on reporter gene expression in tobacco transformants. Plant Mol. Biol. 15: 851-864.
[18.] Marris E. (2008). Water: more crop per drop. Nature 452: 273-277.
[19.] Mayerhofer R., Koncz-kalman Z., Nawrath C., Bakkeren G., Crameri A., Angelis K., Redei G. P., Schell J., Hohn B., and Koncz C. (1991) T-DNA integration: A mode of illegitimate recombination in plants. EMBO Journal 10: 697-704.
[20.] Neill S.J. and Burnett E.C. (1999) Regulation of gene expression during water deficit stress. Plant Growth Regulation 29 : 23-33.
[21.] Osmond C. B., Austin M.P., Berry J. A., Billings W.D., Boyer J. S., Dacey J.W. H., Nobel P. S., Smith S. D., and Winner W.E. (1987). Stress Physiology and the Distribution of Plants. BioScience37: 38-48.
[22.] Pan X., Liu H., Clarke J., Jones J., Bevan M., and Stein L. (2003). ATIDB: Arabidopsis thaliana insertion database. Nucleic Acids Research. 31: 1245-1251.
[23.] Radhamony R. N., Prasad A.M., and Srinivasan R. (2005) T-DNA insertional mutagenesis in Arabidopsis: A tool for functional genomics. Electronic Journal of Biotechnology 8: 82-106.
[24.] Sanderfoot A.A., Kovaleva, V., Bassham, D. C.,andRaikhle, N.V. (2001). Interactions between Syntaxins identify at least five SNARE complexes within the golgi/prevacuolar system of the Arabidopsis cell. Molecular Biology of the Cell 12:3733-43.
[25.] Sessions A., Burke E., Presting G., Aux G., McElver J., Patton D., Dietrich B., Ho P., Bacwaden J., Ko C., Clarke J.D., Cotton D., Bullis D., Snell J., Miguel T., Hutchison D., Kimmerly B., Mitzel T., Katagiri F., Glazebrook J., Law M., and Goff S. A. (2002). A high-throug out Arabidopsis reverse genetics system. Plant Cell 14: 2985-2994
[26.] Shao H. B., Chu L.Y., Jaleel C.A., and Zhao C.X. (2008). Water-deficit stress-induced anatomical changes in higher plants. Comptes Rendus – Biologies 331: 215-225.
[27.] SIGnAL (2000). Arabidopsis sequence indexed T-DNA insertion In http: 11 signal. Salk. Edu/ tdna-fa 95. Html.
[28.] Yang T., Zhang L., Zhang T., Zhang H., Xu S., and An L. (2005). Transcriptional regulation network of cold-responsive genes in higher plants. Plant Science 169 : 987-995.
[29.] Zhu J., Gong Z., Zhang C., Song C.P., Damsz B., Inan G., Koiwa H., Zhu J.K., Hasegawa P.M., and Bressan R.A. (2002). OSM1/SYP61: A syntaxin protein in Arabidopsis controls abscisic acid-mediated and non-abscisic acid-mediated responses to abiotic stress. Plant Cell 14 : 3009-3028.
[30.] The Arabidopsis Genome Initiative (TAGI) (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408 : 796-815.
[31.] QIAGEN (2002) QIAQUICK Spin Handbook available at:
[32.] Van der GRAAFF E., and Hooykaas P. J. J. (1996) Improvements in the transformation of Arabidopsis thaliana C24 leaf-discs by Agrobacteriumtumefaciens. Plant Cell Rep. 15 : 572-577.
