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Profiling of Gene Expression in the Reproductive Organs of Jatropha curcas |
WANG Wei1, WEI Bryan1, PUN Sing1, QING Dong-jin1, WONG Wai-shing1, ZHANG Shi-hua1, WANG Lei2, LI Ning1 |
1. Division of Life Science School of Science The Hong Kong University of Science and Technology Clear Water Bay, Hong Kong SAR, China;
2. School of Life Science Nankai University, Tianjing 300457, China |
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Abstract The expressed sequence tags (ESTs), fragments of mRNA sequences, have been widely used for effective gene discovery and complementation of the genome annotation. It is also beginning to be applied in the fields of phylogenetics, transcript profiling and proteomics recently in combination with quantitative real-time RT-PCR (qRT-PCR). The analysis of genes expression level from reproductive organs of the oilseed-bearing shrub called Jatropha curcas (J. curcas) may reveal some interesting genes related to regulation of lipid biosynthesis. The outcome may someday be used to improve the oil productivity of oil seed-bearing woody plants. Two cDNA libraries were constructed separately. A total of 9289 EST sequences were obtained with an average length of 603 bp (base pair), where as 4502 unique sequences (UniSeqs) were obtained with assembly of these EST sequences, including 1427 contigs (containing more than one reads) and 3075 singletons (containing a single read), respectively. These assembled sequences were annotated with gene names of Gene Ontology (GO) terms. In these annotated UniSeqs, the relative expression level of 50 ESTs was quantified with qRT-PCR in tissues of leaf, flower and seed. It was found that the most abundant 6 ESTs (Contig1452, Contig1482, Contig1510, Contig1514, Contig1534 and Contig1535), which are frequently detected, in two cDNA libraries were also expressed highly in one or two different tissues. These highly expressed genes encode lipid biosynthesis-related proteins or transcript factors. Numerous high quality ESTs for J. curcas genome annotation and increase in the number of sequences deposited in public databases were provided. The qRT-PCR assay has validated the abundance of six interesting ESTs, which were frequently found in cDNA library. Especially, those genes, which are related to both lipid biosynthesis and transcription, have been shown to be actively expressed according to qRT-PCR analysis. These genes may play an important role in regulation of lipid biosynthesis in J. curcas.
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Received: 17 January 2011
Published: 28 June 2011
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Fund: This work is supported by Grants, RPC07/08.SC16, CGPL-2,-3 and-5, CGPL08/09.SC01, SBI08/09.SC08 and GMGS08/09.SC04, from the HKUST R&D Corporation and HKUST |
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[1] Gambino G, Perrone I, Gribaudo I. A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants. Phytochem Anal, 2008, 19(6):520-525.
[2] Ewing B, Green P. Base-calling of automated sequencer traces using phred. Ⅱ. Error probabilities. Genome Res, 1998, 8(3):186-194.
[3] Ewing B, Hillier L, Wendl M C, et al. Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Res, 1998, 8(3):175-185.
[4] Bonfield J K, Smith K, Staden R. A new DNA sequence assembly program. Nucleic Acids Res, 1995, 23(24):4992-4999.
[5] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001, 25(4):402-408.
[6] de La T F, Sampedro J, Zarra I, et al. AtFXG1, an Arabidopsis gene encoding alpha-L-fucosidase active against fucosylated xyloglucan oligosaccharides. Plant Physiol, 2002, 128(1):247-255.
[7] Brick D J, Brumlik M J, Buckley J T, et al. A new family of lipolytic plant enzymes with members in rice, arabidopsis and maize. FEBS Lett, 1995, 377(3):475-480.
[8] Faro C, Ramalho-Santos M, Vieira M, et al. Cloning and characterization of cDNA encoding cardosin A, an RGD-containing plant aspartic proteinase. J Biol Chem, 1999, 274(40):28724-28729.
[9] Pichon M, Courbou I, Beckert M, et al. Cloning and characterization of two maize cDNAs encoding cinnamoyl-CoA reductase (CCR) and differential expression of the corresponding genes. Plant Mol Biol, 1998, 38(4):671-676.
[10] Bouton S, Viau L, Lelievre E, et al. A gene encoding a protein with a proline-rich domain (MtPPRD1), revealed by suppressive subtractive hybridization (SSH), is specifically expressed in the Medicago truncatula embryo axis during germination. J Exp Bot, 2005, 56(413):825-832.
[11] Clemente H S, Pont-Lezica R, Jamet E. Bioinformatics as a tool for assessing the quality of sub-cellular proteomic strategies and inferring functions of proteins: plant cell wall proteomics as a test case. Bioinform Biol Insights, 2009, 3:15-28.
[12] Kader J C. Lipid-transfer proteins in plants. Annu Rev Plant Physiol Plant Mol Biol, 1996, 47:627-654.
[13] Salcedo G, Sanchez-Monge R, az-Perales A, et al. Plant non-specific lipid transfer proteins as food and pollen allergens. Clin Exp Allergy, 2004, 34(9):1336-1341.
[14] Maraschin S F, Caspers M, Potokina E, et al. cDNA array analysis of stress-induced gene expression in barley androgenesis. Physiol Plant, 2006, 127: 535-550.
[15] Honys D, Twell D. Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol, 2004, 5(11):R85.
[16] Whittle C A, Malik M R, Li R, et al. Comparative transcript analyses of the ovule, microspore, and mature pollen in Brassica napus. Plant Mol Biol, 2010, 72(3):279-299. |
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