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Agrobacterium-mediated transf.rmation and assessment of factors influ-encing transgene expression in loblolly pine （Pinus taeda L.）

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Cell Research 2001年第3期11卷 237-243页

作者： tang wei north carolina state university, 2900 ligon sf., raleigh, nc 27607, usa 1north carolina state university 2900 Ligon St. Raleigh NC 27607 USA.

This investigation reports a protocol for transf.r and expression of foreign chimeric genes in loblolly pine (Pinus taeda L.). Transf.rmation was achieved by co-cultivation of mature zygotic embryos with Agrobacterium tumefaciens strain LBA4404 which harbored a binary vector (pBI121) including genes for β-glucuronidase (GUS) and neomycin phosphotransf.rase (NPTII). Factors influencing transgene expression including seed sources of loblolly pine, concentration of bacteria, and the wounding procedures of target explants were investigated. The expression of foreign gene was confirmed by the ability of mature zygotic embryos to produce calli in the presence of kanamycin, by histochemical assays of GUS activity, by PCR analysis, and by Southern blot. The successf.l expression of the GUS gene in different families of loblolly pine suggests that this transf.rmation system is probably useful for the production of the genetically modified conifers.

关键词： Pinus taeda L. Agrobacterum tumefaciens gene transf.r gene expression.

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Recent advances in the molecular genetics of resin biosynthesis and genetic engineering strategies to improve defenses in conifers

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Journal of Forestry Research 2003年第2期14卷 171-179页

作者：唐巍 O-23 E.S.King Village2900 Ligon St.North Carolina State University

Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a monoterpene synthase, a sesquiterpene synthase, and a diterpene synthase. Tree killing bark beetles and their vectored fungal pathogens are the most destructive agents of conifer forests worldwide. Conifers defend against attack by the constitutive and inducible production of oleoresin that accumulates at the wound site to kill invaders and both flush and seal the injury. Although toxic to the bark beetle and fungal pathogen, oleoresin also plays a central role in the chemical ecology of these boring insects. Recent advances in the molecular genetics of terpenoid biosynthesis provide evidence for the evolutionary origins of oleoresin and permit consideration of genetic engineering strategies to improve conifer defenses as a component of modern forest biotechnology. This review described enzymes of resin biosynthesis, structural feathers of genes genomic intron and exon organization, pathway organization and evolution, resin production and accumulation, interactions between conifer and bark beetle, and engineering strategies to improve conifer defenses.

关键词： Genetic engineering strategies Resin biosynthesis Bark beetles Genomics Molecular genetics

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