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Development and Growth Regulation

Identification of Two Brassinosteroids from the Cambial Region of Scots Pine (Pinus silverstris) by Gas Chromatography-Mass Spectrometry, after Detection Using a Dwarf Rice Lamina Inclination Bioassay ¹

Plant Physiology Research Group, Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4, Forestry Canada-Maritimes Region, P. O. Box 4000, Fredericton, New Brunswick, Canada E3B 5P7

A simple and improved dwarf rice (Oryza sativa var Tan-ginbozu) lamina inclination bioassay for brassinosteroids (BRs) was developed based on a previously published method (K Takeno, RP Pharis [1982] Plant Cell Physiol 23: 1275-1281). The assay used 3-day-old intact seedlings, and detection of BR was made more sensitive by synergizing the response to BR with indole-3-acetic acid (IAA). The minimum detectable amount of BR was less than 0.1 nanogram/rice plant (brassinolide equivalents). Purification steps for isolation of BR from tissue scrapings taken from the cambial region of Scots pine (Pinus silverstris) harvested during the period of rapid wood production were guided by this assay. After column chromatography (silica gel and PrepPak C₁₈) and reversed phase C₁₈ high performance liquid chromatography, the biologically active fractions were analyzed by gas chromatography-mass spectrometry (GC-MS) and/or GC-MS-selected ion monitoring. Two BRs, castasterone (major) and brassinolide (minor) were identified. This is the first identification of BR from the cambial region of a conifer.

³ Present address: Department of Plant Protection, Faculty of Agriculture, Tokyo University of Agriculture & Technology, Fuchu, Tokyo 183, Japan.

¹ Supported by a Canadian Forestry Service P.R.U.F. Contract to R. P. P. and National Sciences & Engineering Research Council of Canada Grant A-2585.

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