AJPS  Vol.2 No.5 , November 2011
Expression of Six Chloroplast Genes in Jatropha curcas Callus under Light and Dark Conditions
Abstract: The induction of genes encoded in the open reading frames (ORFs) of chloroplast genomes have been posited to be influenced by ambient light condition. The current study focused on determining which of the six ORFs, encoding the genes ycf 1, ycf 2, psbD (photosystem II), rbcl (Rubisco), matK (Maturase K) and rpoC1 (RNA polymerase) were influenced by light. Characterization of gene expression at the whole plant level and callus stage facilitates the identification of transcripts which are differentially regulated under these environmental conditions. Specificity of these primers was tested against genomic DNA and total RNA. Transcripts of six targeted genes were detected in all three replicates of the green and white callus under light and dark conditions, except for ycf 2 gene in green callus under light. The result showed that a partial transcript of the gene ycf 2 located on the J. curcas chloroplast genome was not detectable using reverse transcription PCR. This finding was then validated using quantitative real-time PCR. The gene was suspected to be post-transcriptionally modified. The transcripts of the remaining five ORFs could be detected using quantitative real-time PCR. Specific transcripts can be identified for application as biomarkers for selection of callus for plantlet regeneration.
Cite this paper: nullW. Yong, C. Chew and K. Rodrigues, "Expression of Six Chloroplast Genes in Jatropha curcas Callus under Light and Dark Conditions," American Journal of Plant Sciences, Vol. 2 No. 5, 2011, pp. 650-656. doi: 10.4236/ajps.2011.25077.

[1]   W. M. J. Achten, L. R. Nielsen, R. Aerts, A. G. Lengkeek, E. D. Kjaer, A. Trabucco, J. K. Hansen, W. H. Maes, L. Graudal, F. K. Akinnifesi and B. Muys, “Towards Domestication of Jatropha curcas,” Biofuels, Vol. 1, No. 1, 2010, pp. 91-107. doi:10.4155/bfs.09.4

[2]   J. Gressel, “Transgenics Are Imperative for Biofuel Crops,” Plant Science, Vol. 174, No. 3, 2008, pp. 246-263. doi:10.1016/j.plantsci.2007.11.009

[3]   R. de la Pena and J. Hughes, “Improving Vegetable Pro- ductivity in a Variable and Changing Climate,” SAT eJournal, Vol. 4, 2007.

[4]   T. Murashige and F. Skoog, “A Revised Medium for Ra- pid Growth and Bioassays with Tobacco Tissue Cultures,” Physiologia Plantarum, Vol. 15, No. 3, 1962, pp. 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x

[5]   J. J. Doyle and J. L. Doyle, “A Rapid DNA Isolation Procedure for Small Quantities of Fresh Leaf Tissue,” Phytochemical Bulletin, Vol. 19, 1987, pp. 11-15.

[6]   J. Sambrook and D. W. Russell, “In vitro Amplification of DNA by the Polymerase Chain Reaction,” In: J. Sam- brook and D. W. Russell, Eds., Molecular Cloning: A Laboratory Manual, 3rd Edition, Cold Spring Harbor Laboratory Press, New York, 2001, pp. 8.1-8.113.

[7]   S. F. Altschul, W. Gish, W. Miller, E. W. Myers and D. J. Lipman, “Basic Local Alignment Search Tool,” Journal of Molecular Biology, Vol. 215, 1990, pp. 403-410.

[8]   I. Shahzadi, R. Ahmed, A. Hassan and M. M. Shah, “Op- timization of DNA Extraction from Seeds and Fresh Leaf Tissues of Wild Marigold (Tagetes minuta) for Poly- merase Chain Reaction Analysis,” Genetics and Molecu- lar Research, Vol. 9, No. 1, 2010, pp. 386-393. doi:10.4238/vol9-1gmr747

[9]   S. T. Ying and F. Q. Zaman, “DNA Extraction from Ma- ture Oil Palm Leaves,” Journal of Oil Palm Research, Vol. 18, 2006, pp. 218-224.

[10]   D. Dhakshanamoorthy and R. Selvaraj, “Extraction of Genomic DNA from Jatropha sp. Using Modified CTAB Method,” Romanian Journal of Biology—Plant Biology, Vol. 54, 2009, pp. 117-125.

[11]   T. Philips, “Regulation of Transcription and Gene Ex- pression in Eukaryotes,” Nature Education, Vol. 1, No. 1, 2008.

[12]   K. M. Neubig, W. M. Whitten, B. S. Carlsward, M. A. Blanco, L. Endara, N. H. Williams and M. Moore, “Phylogenetic Utility of ycf 1 in Orchids: A Plastid Gene More Variable Than matK,” Plant Systematics and Evolution, Vol. 277, No. 1-2, 2009, pp. 75-84. doi:10.1007/s00606-008-0105-0

[13]   L. D. Crosby and C. S. Criddle, “Gene Capture and Ran- dom Amplification for Quantitative Recovery of Ho- mologous Genes,” Molecular and Cellular Probes, Vol. 21, No. 2, 2007, pp. 140-147. doi:10.1016/j.mcp.2006.09.003

[14]   S. Wicke and D. Quandt, “Universal Primers for the Am- plification of the Plastid trnK/matK Region in Land Plants,” Anales del Jardin Botanico de Madrid, Vol. 66, 2009, pp. 285-288. doi:10.3989/ajbm.2231

[15]   S. Pafundo, M. Gulli and N. Marmiroli, “SYBR? Green- ERTM Real-Time PCR to Detect Almond in Traces in Processed Food,” Food Chemistry, Vol. 116, No. 3, 2009, pp. 811-815. doi:10.1016/j.foodchem.2009.03.040

[16]   R. M. Twyman, “Growth and Development: Control of Gene Expression, Posttranscriptional Regulation,” In: B. Thomas, D. J. Murphy and B. Murray, Eds., Encyclopedia of Applied Plant Sciences, Elsevier Science, London, 2003, pp. 549-557.

[17]   M. Floris, H. Mahgoub, E. Lanet, C. Robaglia and B. Menand, “Post-Transcriptional Regulation of Gene Expression in Plants during Abiotic Stress,” International Journal of Molecular Sciences, Vol. 10, No. 7, 2009, pp. 3168- 3185. doi:10.3390/ijms10073168

[18]   M. Durante, M. Pieretti and R. Bernardi, “Chloroplastic ycf 2 Gene Expression in Stressed Plants,” Proceedings of the 53rd Italian Society of Agricultural Genetics Annual Congress, Torino, 16-19 September 2009.