Transcription profiles analysis of genes encoding 1-deoxy-D-xylulose 5-phosphate synthase and 2C-methyl-D-erythritol 4-phosphate synthase in plaunotol biosynthesis from Croton stellatopilosus.
Genes encoding 1-deoxy-D-xylulose 5-phosphate synthase (DXS; EC 2.2.1.7) and 2C-methyl-D-erythritol 4-phosphate synthase (MEPS; EC 1.1.1.267), the first two enzymes in the deoxyxylulose phosphate (DXP) pathway, were cloned from young leaves of Croton stellatopilosus, and designated as 1-deoxy-D-xylulose 5-phosphate synthase (CSDXS) and 2C-methyl-D-erythritol 4-phosphate synthase (CSMEPS), respectively. Analysis of deduced amino acid sequences of the CSDXS and the CSMEPS confirmed their nucleotide sequences as they shared high identities to other known DXSs and MEPSs in higher plants. Physiological roles of the CSDXS and the CSMEPS were determined for the mRNA expressions in leaves, twigs and roots. Transcription profiles analyses of the CSDXS and the CSMEPS genes were investigated using semi-quantitative RT-PCR technique. Relative intensities of the CSDXS and the CSMEPS expressions to house-keeping gene (18S rRNA) were calculated. The results indicated that the levels of mRNAs expressions of the CSDXS and the CSMEPS were high in leaves and twigs. This evidence was in line with the high content of plaunotol, accumulated in leaves and twigs. Neither the CSDXS nor the CSMEPS were expressed in roots, where plaunotol was not detected. From this study, it can be concluded that plaunotol is biosynthesized in the chloroplastic tissue and regulated by the CSDXS and the CSMEPS.
Wungsintaweekul
Juraithip
J
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand. juraithip.w@psu.ac.th
Sirisuntipong
Tanawan
T
Kongduang
Damrong
D
Losuphanporn
Thanaisawan
T
Ounaroon
Anan
A
Tansakul
Pimpimon
P
De-Eknamkul
Wanchai
W
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D002478:Cells, Cultured; D002849:Chromatography, Gas; D003001:Cloning, Molecular; D029802:Croton / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification; D004224:Diterpenes; D005233:Fatty Alcohols / Q000378:metabolism*; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000096:biosynthesis* / Q000235:genetics; D018515:Plant Leaves / Q000201:enzymology / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology / Q000378:metabolism; D018547:Plant Stems / Q000201:enzymology / Q000378:metabolism; D015202:Protein Engineering; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014166:Transferases / Q000096:biosynthesis* / Q000235:genetics
D000900:Anti-Bacterial Agents; D017931:DNA Primers; D018076:DNA, Complementary; D004224:Diterpenes; D005233:Fatty Alcohols; D012333:RNA, Messenger; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase; C400183:4-diphosphocytidyl-2-C--methylerythritol synthase; D009713:Nucleotidyltransferases; C047686:plaunotol