tailieunhanh - Báo cáo khoa học: "Study of endogenous plant growth Douglas fir I. Cytokinin analysis"

Tuyển tập các báo cáo nghiên cứu về lâm nghiệp được đăng trên tạp chí lâm nghiệp quốc tế đề tài: "Study of endogenous plant growth Douglas fir I. Cytokinin analysis. | 256s Ann. Sci. For. 1989 46 suppl. 256s-258s Forest Tree Physiology E. Dreyer etal. eds. Elsevier INRA Study of endogenous plant growth substances in Douglas fir I. Cytokinin analysis N. Imbault1 p. Doumas2 c. Joseph1 and M. Bonnet-Masimbert2 1 Laboratoire des Composes Phẽnoliques Université d Orleans BP 6769 45067 Orleans Cedex 02 and 2 INRA Station d Amelioration des Arbres Forestiers Ardon 45160 Olivet France Introduction To ascertain the part played by a natural substance in a biological phenomenon it is necessary to follow the endogenous evolution of this compound during the induction of the process. This is a real problem with plant growth substances PGS . Indeed their very low concentrations in tissues make PGS difficult to quantify. Because of their sensitivity and specificity immunological methods have been adapted to the analysis of PGS and enable in some cases measurements at the level of a single organ as reported for principally herbaceous species Weiler 1984 . In this paper some of their applications to the woody plant Douglas fir Pseudotsuga menziesii Mirb. are presented purification by immunoaffinity chromatography IAC and measurement by an enzyme-linked immunosorbent assay ELISA or a radioimmunoassay RIA . Materials and Methods Material The study was performed on sexual buds of Douglas fir. Cytokinin isolation Cytokinins were extracted with 80 methanol in phosphate buffer pH . After concentration the extracts were passed through a diethylami-noethyl-cellulose column and purified either on an immunoaffinity IA column as described below or on an octadecylsilica one. Cytokinins were then separated by high-performance liquid chromatography HPLC using a reverse phase column MacDonald etal. 1981 and measured either by uv absorption or by ELISA or RIA as reported below . Immunological methods For IAC and ELISA procedures monoclonal antibodies were raised against cytokinins conjugated to bovine serum albumin MacDonald and Morris 1985 . IA columns of 1

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