Abstract
Cell-free extracts from Pinus ponderosa Lawson (ponderosa pine) and Pinus sylvestris L. (Scotch pine) wood exhibited high levels of monoterpene synthase (cyclase) activity, whereas bark extracts of these species contained no detectable activity, and they inhibited cyclase activity when added to extracts from wood, unless polyvinylpyrrolidone was included in the preparation. The molecular mass of the polyvinylpyrrolidone added was of little consequence; however, polyvinylpolypyrrolidone (a cross-linked insoluble form of the polymer) was ineffective in protecting enzyme activity. Based on these observations, methods were developed for the efficient extraction and assay of monoterpene cyclase activity from conifer stem (wood and bark) tissue. The level of monoterpene cyclase activity for a given conifer species was shown to correlate closely with the monoterpene content of the oleoresin and with the degree of anatomical complexity of the specialized resin-secreting structures. Cyclase activity and monoterpene content were lowest in the stems of species containing only isolated resin cells, such as western red cedar (Thuja plicata D. Don). Increasing levels of cyclase activity and oleoresin monoterpenes were observed in advancing from species with multicellular resin blisters (true firs [Abies]) to those with organized resin passages, such as western larch (Larix occidentalis Nutt.), Colorado blue spruce (Picea pungens Engelm.) and Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco). The highest levels of cyclase activity and oleoresin monoterpenes were noted in Pinus species that contain the most highly developed resin duct systems. The relationship between biosynthetic capacity, as measured by cyclase activity, monoterpene content, and the degree of organization of the secretory structures for a given species, may reflect the total number of specialized resin-producing cells per unit mass of stem tissue.
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- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Cori O. Terpene biosynthesis: utilization of neryl pyrophosphate by an enzyme system from Pinus radiata seedlings. Arch Biochem Biophys. 1969 Dec;135(1):416–418. doi: 10.1016/0003-9861(69)90558-x. [DOI] [PubMed] [Google Scholar]
- Croteau R., Gurkewitz S., Johnson M. A., Fisk H. J. Biochemistry of Oleoresinosis : Monoterpene and Diterpene Biosynthesis in Lodgepole Pine Saplings Infected with Ceratocystis clavigera or Treated with Carbohydrate Elicitors. Plant Physiol. 1987 Dec;85(4):1123–1128. doi: 10.1104/pp.85.4.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haissig B. E., Schipper A. L., Jr Easy extraction of enzymes from small amounts of woody tissue. Anal Biochem. 1972 Jul;48(1):129–146. doi: 10.1016/0003-2697(72)90177-7. [DOI] [PubMed] [Google Scholar]
- Karp F., Croteau R. Evidence that sabinene is an essential precursor of C(3)-oxygenated thujane monoterpenes. Arch Biochem Biophys. 1982 Jul;216(2):616–624. doi: 10.1016/0003-9861(82)90251-x. [DOI] [PubMed] [Google Scholar]
- Loomis W. D. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Methods Enzymol. 1974;31:528–544. doi: 10.1016/0076-6879(74)31057-9. [DOI] [PubMed] [Google Scholar]
- Satterwhite D. M., Croteau R. B. Applications of gas chromatography to the study of terpenoid metabolism. J Chromatogr. 1988 Oct 28;452:61–73. doi: 10.1016/s0021-9673(01)81437-0. [DOI] [PubMed] [Google Scholar]