Temporal and Spatial Regulation of Cyanogenesis in Plants

植物发氰的时空调控

• 批准号: 9218929
• 负责人: Jonathan Poulton
• 金额: $ 29.1万
• 依托单位: University of Iowa
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-02-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9218929&HistoricalAwards=false
• 关键词: Temporal; Spatial; Regulation; Cyanogenesis; Plants

Rosaceous stone fruits (Prunus spp.) are among many commercially important crops causing cyanide poisoning in humans and livestock through their release of hydrogen cyanide from cyanogenic glycosides by endogenous enzymes. To understand better the biochemistry and physiology of plant cyanogenesis, the metobolism of the cyanoglycosides (R)-prunasin and (R)-amygdalin will be investigated in black cherry (Prunus serotina) using biochemical, immunocytochemical and molecular approaches. Levels of prunasin, amygdalin, two biosynthetic enzymes (UDPG:mandelonitrile glucosyltransferase (GT I) and UDPG:prunasin glucosyltransferase (GT II), and three catabolic enzymes (amygdalin hydrolase (AH), prunasin hydrolase (PH) and mandelonitrile lyase (MDL)) will be assayed during embryogenesis. The distribution of GT I and II and of amygdalin within seeds will be determined by immunocytochemistry and FT-IR microspectroscopy, respectively. Using previously raised polyclonal antibodies, an existing black cherry cDNA library will be screened to obtain AH, PH and MDL cDNA clones. These clones will be utilized to begin examining the temporal and spatial expression of these genes during embryogenesis by Northern blot analysis and in situ hybridization techniques. Whether germinating seedlings utilize amygdalin as a nitrogen source will be examined by measuring the levels and localization of the aforementioned cyanoglycosides and enzymes, along with the HCN detoxification enzyme Beta-cyanoalanine synthase, in various seedling parts.

蔷薇核果（Prunus spp.）是许多商业上 导致人类和牲畜氰化物中毒的重要作物 通过从氰化物中释放出氰化氢 通过内源性酶合成糖苷。 更好地了解 植物成氰的生理生化、代谢 氰基糖苷类（R）-prunasin和（R）-苦杏仁苷将 研究了黑樱桃（Prunus damnina）的生化、 免疫细胞化学和分子方法。 prunasin的水平， 苦杏仁苷，两种生物合成酶（UDPG：扁桃腈 葡糖基转移酶（GT I）和UDPG：prunasin葡糖基转移酶 (GT II）和三种分解代谢酶（苦杏仁苷水解酶（AH）， prunasin水解酶（PH）和扁桃腈裂解酶（MDL））将被 在胚胎发生期间测定。 GT I和II的分布， 将通过免疫细胞化学测定种子内苦杏仁苷的含量 和FT-IR显微光谱。 使用以前提出的 多克隆抗体，现有的黑樱桃cDNA文库将 筛选获得AH、PH和MDL cDNA克隆。 这些克隆 将被用来开始检查时间和空间 通过北方杂交检测这些基因在胚胎发生过程中的表达 分析和原位杂交技术。 发芽的幼苗是否利用苦杏仁甙作为氮素 来源将通过测量的水平和本地化进行审查， 上述氰基糖苷和酶，沿着HCN 解毒酶β-氰基丙氨酸合酶，在各种 幼苗部分。