A New Farnesyl Diphosphate Synthase Gene from Taxus media Rehder:Cloning,Characterization and Functional Complementation

Farnesyl dlphosphate synthase （FPS; EC 2.5.1.10） catalyzes the production of 15-carbon farnesyl dlphosphate which Is a branch-point Intermediate for many terpenoids. This reaction Is considered to be a ratelimiting step In terpenold biosynthesis. Here we report for the first time the cloning of a new full-length cDNA encoding farnesyl dlphosphate synthase from a gymnosperm plant species, Taxus media Rehder, designated as TmFPS1. The full-length cDNA of TmFPS1 （GenBank accession number： AY461811） was 1 464 bp with a 1 056-bp open reading frame encoding a 351-amino acid polypeptlde with a calculated molecular weight of 40.3 kDa and a theoretical pl of 5.07. Biolnformatlc analysis revealed that TmFPS1 contained all five conserved domains of prenyltransferases, and showed homology to other FPSs of plant origin. Phylogenetlc analysis showed that farnesyl dlphosphate synthases can be divided Into two groups： one of prokaryotic origin and the other of eukaryotic origin. TmFPS1 was grouped with FPSs of plant origin. Homologybased structural modeling showed that TmFPS1 had the typical spatial structure of FPS, whose most prominent structural feature Is the arrangement of 13 core helices around a large central cavity In which the catalytic reaction takes place. Our blolnformatic analysis strongly suggests that TmFPS1 is a functional gene. Southern blot analysis revealed that TmFPS1 belongs to a small FPSgene family in T. media. Northern blot analysis indicated that TmFPS1 is expressed in all tested tissues, Including the needles, stems and roots of T. media. Subsequently, functional complementatlon with TmFPS1 in a FPS-deflclent mutant yeast demonstrated that TmFPS1 did encode farnesyl dlphosphate synthase, which rescued the yeast mutant. This study will be helpful In future Investigations aiming at understanding the detailed role of FPS In terpenold biosynthesis flux control at the molecular genetic level.

Brassica napus L. Homeodomain Leucine-Zipper Gene BnHB6 Responds to Abiotic and Biotic Stresses

A homeodomain leucine-zipper （HD-Zip） gene BnHB6 （GenBank accession No. AY336103） was isolated from oilseed rape （Brassica napus L.） following drought treatment through rapid amplification of cDNA ends （RACE）. The full-length cDNA of BnHB6 was 1 611 bp and contained a 936-bp open reading frame encoding 311 amino acids. Sequence analysis indicated that BnHB6 belonged to the HD-Zip I subfamily. High-stringency Southern boltting analysis showed that BnHB6 appeared in rape as a single copy but had homologous genes. Semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR） analysis revealed that BnHB6 was expressed in several tissues tested under control conditions, but that expression was significantly upregulated in shoots by mannitol, NaCI, cold treatment, anaerobic culture, wounding, H2O2, abscisic acid （ABA）, and salicylic acid （SA） treatments, but not by ultraviolet treatment. Further RT- PCR analysis revealed that BnHB6 was a late-responsive gene, the expression of which was not activated by NaCI, cold treatment, H2O2, ABA, and SA at an early time point （20 min） of treatment in the shoot. However, after a certain period of treatment, the induced expression culminated and then declined until the next peak occurred. Tissue-specific analysis revealed that BnHB6 was expressed at certain levels in the roots, shoots, and flowers, and the roots were found to respond to the osmotic stimuli more rapidly than shoots to increase the expression of BnHB6. The present study implies that BnHB6 plays a positive role as a regulator of biotic and abiotic stresses on growth during seedling establishment.