| Phospholipase D alpha1(PLDal) plays important roles in the processes of cell remolding&proliferation, stress and phytohormone responses in plants, and is a key enzyme involving in the wound signal transduction and the wound-induced jasmonic acid (JA) production. JA biosynthesis and JA-mediated signal transduction in the laticifers of Hevea brasiliensis (rubber tree) remain unclear so far. In the research, the expression pattern of HbPLDal, a PLDα1gene in the latex of the mature rubbe trees (Clone reyuan7-33-97), was investigated, and the data would facilitate unraveling the signaling pathway of JA and its regulation of the rubber biosynthesis in rubber trees. The main results were summarized as follows:1. HbPLDal and its promoter were cloned based on the EST sequences using RACE and Genome Walking. The full-length cDNA of HbPLDα1gene was2870bp which contained an open reading frame (ORF) of2427bp encoding808amino acids,227bp of5’ UTR and216of3’UTR. The molecular mass (MW) of the putative HbPLDal protein is a92.2kDa with a theoretical p1of5.52. The promoter of HbPLDal gene is1559bp long.2. Bioinformatic analysis shows that HbPLDal mainly contain four conserved domains with a C2domain located at N-terminal, two active sites (PLDc) and a PLD C domain located at C-terminal, which indicates that HbPLDal has the similar conserved domains of plant PLD with those of other plants, and the highest identity with PLDα1from Ricinus communis and Jatropha curcas. The promoter of HbPLDal includes not only the common cis-acting elements of TATA and CAAT box, but also cis-acting elements responsive to phytohormones (JA, ABA, GB) and environmental stresses.3. The gene expression of HbPLDal was regulated by the stimulation of exogenous JA but not of the treatment of ethephon.4. Western blotting analysis demonstrated that HbPLDal was mainly expressed on the small rubber particles (SRPs) but not on the large rubber particles (LRPs) in the latex of rubber trees. |
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