Response of Phenylethanoid Systems to Photoperiod in Rose Flower Petals

玫瑰花瓣中苯乙醇系统对光周期的响应

• 批准号: 16310149
• 负责人: WATANABE Naoharu
• 金额: $ 9.79万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16310149/
• 关键词: 2-phenylethanol; phenylacetaldehyde; decarboxy, ase; aldoreductase; β-glucosidase; biosynthesis; rose flower; LED; ケトレダクターゼ; バラ花弁; 明暗応答; フェニルアラニン

We have found that the endogenous levels of molecules, such as enzymes and volatile components, involved in the scent emission were changed by the photoperiod. In this research we focused our research to elucidate the biosynthetic pathways of 2-phenylethanol (2PE) and its β-D-glucopyranoside (2PEG). We also tried to clarify the phenylethanoid systems, such as volatile compounds, their glucosides, enzymes, and the genes, being involved in the scent emission from the flower petals of roses.1. Identification of PLP-dependent aromatic amino acid decarboxylase (AADC) and phenvlacetaldehvde reductase. (PAR)Phenylacetaldehyde (PAId), the direct precursor o 2PE, was produced from L-phenylalanine (Phe). PLP-dependent AADC directly produced PAId together with NH_3 and H_2O_2 under aerobic conditions. The activity of rose petal AADC to yield PAId showed nine times higher activity toward L-Phe than toward its D-isomer. PAId was subsequently converted to 2PE by the action of PAR. The PAR showed spe … More cificity toward several volatile aldehydes, whereas ketone groups of the volatile compounds were not reduced at all.2. Isolation and characterization of β-glucosidase, involved in the hydrolysis of 2PEG to produce 2PE.We have successfully isolated β-glucosidase from the flower petals of roses. The partial peptide sequences were characterized by TOF-MS/MS after the partial hydrolysis of the enzyme. Based on the analysis of the sequences, we have cloned the full-length β-glucosidase using the Rose EST sequence, and expressed the gene in the insects cell systems. The rose petal β-glucosidase showed a specificity toward β-glucopyranoside of several plant volatile compounds, whereas the activity toward the α-glucopyranosides, galactopyranosides, xylopyranosides, were quite low.3. Response of 2PE emission toward Photoperiod and the effects of LED-colorsAlthough we could trace the change in the 2PE emission from the flowers of intact rose plants throughout 12L-12D-photoperioc, the amount of 2PE emitted from each flower was fluctuated. Thus we have developed the new methods to trace the change in the 2PE emission by using flower petals of roses. The diurnal emission of 2PE was observed for at least three days. The similar patter was observed when Red LED was used, although blue and green LED did not have such effects. As a continuing research, the regulation of 2PE emission will be examined based on mRNA expression, enzymatic activity and change in the levels of several small molecules. involved in the phenylethanoid systems. Less

我们已经发现，内源性分子水平，如酶和挥发性成分，参与气味的排放光周期的变化。本研究旨在阐明2-苯基乙醇（2 PE）及其β-D-吡喃葡萄糖苷（2 PEG）的生物合成途径。我们还试图澄清苯乙醇系统，如挥发性化合物，其糖苷，酶和基因，参与玫瑰花瓣的气味排放。PLP依赖性芳香族氨基酸脱羧酶（AADC）和苯乙醛还原酶的鉴定。（PAR）苯乙醛（PAId）是2 PE的直接前体，由L-苯丙氨酸（Phe）产生。在有氧条件下，PLP依赖的AADC与NH_3和H_2O_2一起直接产生PAId。玫瑰花瓣AADC产生PAId的活性显示对L-Phe的活性比对其D-异构体的活性高9倍。PAId随后通过PAR的作用转化为2 PE。PAR显示， ...更多信息 对几种挥发性醛类化合物的选择性降低，而挥发性化合物的酮基没有减少.β-葡萄糖苷酶的分离与性质研究从玫瑰花瓣中分离得到β-葡萄糖苷酶，该酶参与了2-PEG水解生成2-PE的过程。在酶的部分水解后，通过TOF-MS/MS表征部分肽序列。在序列分析的基础上，利用Rose EST序列克隆了β-葡萄糖苷酶基因的全长，并在昆虫细胞中进行了表达。玫瑰花瓣β-葡萄糖苷酶对几种植物挥发性化合物中的β-葡萄糖苷具有专一性，而对α-葡萄糖苷、半乳糖苷、木糖苷的酶活性较低. 2 PE释放对光周期的响应和LED颜色的影响虽然我们可以跟踪完整的玫瑰花植株在整个12 L-12 D光周期中2 PE释放的变化，但每朵花的2 PE释放量是波动的。因此，我们已经开发了新的方法来跟踪的变化，2 PE排放的玫瑰花瓣。2 PE的日排放至少观察了三天。当使用红色LED时观察到类似的图案，尽管蓝色和绿色LED没有这样的效果。作为一个持续的研究，2 PE排放的调节将根据mRNA表达，酶活性和几个小分子水平的变化进行检查。参与了苯乙醇系统。少