Kim S K, Abe H, Little C H, Pharis R P
Plant Physiology Research Group, Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Plant Physiol. 1990 Dec;94(4):1709-13. doi: 10.1104/pp.94.4.1709.
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 C(18)) and reversed phase C(18) 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.
基于之前发表的方法（K Takeno，RP Pharis [1982]《植物细胞生理学》23：1275 - 1281），开发了一种简单且改进的用于检测油菜素甾醇（BRs）的矮化水稻（Oryza sativa var Tan - ginbozu）叶片倾斜生物测定法。该测定法使用3日龄的完整幼苗，通过使对BR的反应与吲哚 - 3 - 乙酸（IAA）协同作用，提高了对BR检测的灵敏度。BR的最低可检测量小于0.1纳克/水稻植株（油菜素内酯当量）。在此测定法的指导下，对快速木材生产期间收获的苏格兰松（Pinus silverstris）形成层区域的组织刮屑进行BR分离的纯化步骤。经过柱色谱（硅胶和PrepPak C(18)）和反相C(18)高效液相色谱后，通过气相色谱 - 质谱联用（GC - MS）和/或GC - MS选择离子监测对生物活性馏分进行分析。鉴定出两种BR，即表油菜素内酯（主要）和油菜素内酯（次要）。这是首次从针叶树的形成层区域鉴定出BR。
