發(fā)布時間：2018-03-19 05:00

  本文選題：洋水仙　切入點：香氣成分　出處：《福建農(nóng)林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：洋水仙(Narissus spp.)花大色艷,花型多變,葉姿瀟灑飄逸,是世界著名的觀賞花卉。洋水仙栽培品種眾多,但是多數(shù)品種花香較淡或者無香,品種間花香差異較大。為闡明洋水仙不同品種間花香存在差異的原因及其香氣形成機制,以濃香型洋水仙'新娘皇冠'和淡香型洋水仙'胡德山'為材料,鑒定了兩種洋水仙的香氣成分,確定了洋水仙致香的關(guān)鍵成分——乙酸苯甲酯,通過分析乙酸苯甲酯合成關(guān)鍵酶(醇�；D(zhuǎn)移酶)、乙酸苯甲酯合成前體(苯甲醇)與乙酸苯甲酯之間的關(guān)系,結(jié)合醇�；D(zhuǎn)移酶基因的克隆及表達(dá)分析,從生理生化及分子生物學(xué)水平,綜合探討了不同品種洋水仙花香存在差異的原因,初步研究了洋水仙香氣形成的機制,為洋水仙花香育種研究提供了理論參考。主要研究結(jié)果如下:1.利用頂空固相微萃取(HS-SPME)和氣相色譜-質(zhì)譜聯(lián)用(GC-MS)技術(shù),分析了洋水仙花開放過程中香氣成分及含量的變化,結(jié)果表明:濃香型品種'新娘皇冠'主要的香氣物質(zhì)類別是萜烯類、酯類和醇類,主要的香氣成分是(z)-β-羅勒烯、乙酸苯甲酯、苯甲酸甲酯、別羅勒烯、桉葉油醇,特征香氣成分中貢獻(xiàn)最大的是乙酸苯甲酯和(z)-β-羅勒烯。淡香型品種'胡德山'主要的香氣物質(zhì)類別是萜烯類和醇類,主要的香氣成分是(z)-β-羅勒烯、別羅勒烯、β-月桂烯、對苯二甲醚、桉葉油醇,特征香氣成分中貢獻(xiàn)最大的是(z)-β-羅勒烯。濃香型與淡香型洋水仙香氣的差異主要是酯類化合物種類及含量的差異,酯類化合物對洋水仙香氣形成有著重要的影響,乙酸苯甲酯是洋水仙致香的關(guān)鍵成分,決定洋水仙花香的濃淡以及是否具有甜香的特征。2.利用DTNB比色法研究了洋水仙花開放過程中醇酰基轉(zhuǎn)移酶(AAT)活性的變化,兩種洋水仙AAT酶活性隨著花的開放呈現(xiàn)先升后降的趨勢,從花蕾期到盛花期,酶活性逐漸升高,到衰敗期又有所下降。濃香品種'新娘皇冠'花開放過程中酯類化合物的含量與AAT酶活性呈現(xiàn)正相關(guān),淡香品種'胡德山'酯類物質(zhì)的含量與AAT酶活性之間并無明顯的相關(guān)性。3.通過乙酸乙酯溶劑萃取法結(jié)合氣相色譜-質(zhì)譜聯(lián)用技術(shù),分析了洋水仙花開放過程中內(nèi)源苯甲醇含量的變化。濃香品種'新娘皇冠'花開放的4個花期均萃取到苯甲醇,其含量與乙酸苯甲酯的含量呈現(xiàn)正相關(guān),而淡香品種'胡德山' 4個花期均沒有萃取到苯甲醇,苯甲醇的缺失是'胡德山'不能合成乙酸苯甲酯的原因。乙酸苯甲酯的合成受苯甲醇和AAT酶活性的共同影響,苯甲醇供應(yīng)不足或AAT酶活性較低均會使乙酸苯甲酯的合成受阻。在苯甲醇底物供應(yīng)充足的基礎(chǔ)上,同時擁有較高的AAT酶活性,是大量合成乙酸苯甲酯的關(guān)鍵。4.通過外源苯甲醇處理淡香品種'胡德山'花開放過程中不同花期的花朵,研究了苯甲醇處理之后'胡德山'合成乙酸苯甲酯的情況。結(jié)果表明:在外源苯甲醇充足的情況下,'胡德山' 4個花期均萃取到一定含量的乙酸苯甲酯,說明淡香型洋水仙也具有合成乙酸苯甲酯的能力。研究解決淡香型洋水仙自身苯甲醇底物供應(yīng)不足的辦法,是使淡香型洋水仙具有濃郁甜香的關(guān)鍵。5.利用RT-PCR與RACE技術(shù),從'新娘皇冠'和'胡德山'花中分別克隆得到一條醇�；D(zhuǎn)移酶(AAT)基因,分別命名為NtAAT-BC和NtAAT-MD。兩基因間氨基酸同源性達(dá)到98%,同屬中保守性較高,與其他科屬物種的AAT氨基酸同源性相對偏低,NtAAT-BC和NtAAT-MD均含有不同物種間AAT基因普遍存在且高度保守的HXXXD和FGWG基序,可以初步確定已成功從兩種洋水仙中分離出AAT基因。6.采用實時熒光定量PCR技術(shù)分析了兩種洋水仙花開放過程中醇�；D(zhuǎn)移酶(AAT)基因的表達(dá)模式,結(jié)果表明:AAT基因在洋水仙不同品種和花開放的不同時期存在著表達(dá)差異。'新娘皇冠' AAT基因的表達(dá)模式與AAT酶活性、酯類化合物的變化趨勢一致,'新娘皇冠' AAT基因很可能參與了 '新娘皇冠'酯類化合物的合成調(diào)控。AAT基因的差異表達(dá)可能影響洋水仙不同品種間酯類物質(zhì)的合成。
[Abstract]:Narcissus (Narissus spp.) large showy flowers, flowers and Ye Zi, chic and elegant, is the world famous ornamental flower Narcissus cultivation. Many varieties, but most varieties of flowers is light or no incense, flowers larger among varieties. Differences between different varieties of Narcissus in order to clarify the differences in the formation mechanism of floral deposit and its reasons aroma to Luzhou Narcissus' bride crown 'and pink Narcissus' Hood' as material, two kinds of aroma components of Narcissus were identified and determined by the key components of Narcissus fragrance - benzyl acetate, through the analysis of acetic acid benzene ester synthesis key enzyme (AAT), benzyl acetate the synthesis of precursor (benzyl alcohol) and benzyl acetate, combined with the cloning and expression analysis of alcohol acyltransferase gene, from the physiology and biochemistry and molecular biology level, a comprehensive discussion of the different varieties of ocean water Xian differences in flowers A preliminary study on the causes, mechanism of Narcissus aroma formation, provides a theoretical reference for narcissus flower breeding. The main results are as follows: 1. using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) technology, analyzes the changes of aroma components and contents of Narcissus in the process of opening up show: Luzhou 'crown' main varieties bride aroma categories are terpenes, esters and alcohols, the main aroma components of the (z) - beta ocimene, benzyl acetate, methyl benzoate, alloocimene, eucalyptus oil alcohol, with characteristic aroma components was the largest of benzyl acetate and (z) - beta ocimene. Pink variety 'Hood' main aroma categories are terpenes and alcohols, the main aroma components of the (z) - beta ocimene, alloocimene, myrcene, two of benzene ether, eudesmol, characteristic aroma components the contribution of the The (z) - beta ocimene. Differences in Luzhou and pink Narcissus aroma is mainly different kinds and contents of ester compounds, ester compounds have important influence on the formation of Narcissus aroma, benzyl acetate is a key component of Narcissus by incense, Narcissus flowers shade and decision whether.2. has the characteristics of sweet use DTNB Narcissus in the opening process of alcohol acyltransferase Colorimetry (AAT) activity changes of two kinds of Narcissus AAT activity with flower decreased, from bud stage to flowering, enzyme activity gradually increased, and declined to a period of decline at present positive correlation content and AAT activity of Luzhou varieties' crown 'bride flowers esters in the process between the content and AAT activity of fragrant varieties' Hood' esters of.3. have no obvious correlation with ethyl acetate Chromatography-mass spectrometry method of solvent extraction combined with gas, analyzes the changes of endogenous Benzenemethanol Narcissus in the opening process. 4 flowering fragrant varieties' bride 'crown of flower opening were extracted by benzyl alcohol, are positively related to the content and content of benzyl acetate, and fragrant varieties' Hood' 4 flowering were not extracted by benzyl alcohol, benzyl alcohol is lack of reason 'Hood' can't synthesis of benzyl acetate. The synthesis of benzyl acetate together by benzyl alcohol and AAT enzyme activity, benzyl alcohol shortage or AAT low enzyme activity can cause the synthesis of acetic acid benzylester blocked in benzene methanol substrate supply adequate basis, the activity of AAT and have high, different flowers key.4. synthesis of phenyl acetate methyl benzene methanol treated by exogenous fragrant varieties' Hood 'flowers in the process of benzene Situation of Mount Hood 'synthesis of phenyl acetate methyl alcohol treatment after'. The results showed that: in the exogenous benzyl alcohol is sufficient, the 'Hood' 4 stage was extracted into ethyl methyl benzene content, indicating that pink Narcissus is synthesis of benzyl acetate. To study and solve the pink Narcissus itself Benzenemethanol substrate supply problems, is the key to.5. pink Narcissus with rich sweet using RT-PCR and RACE technology, from the 'bride crown' and 'Hood' flowers were cloned from an alcohol acyltransferase (AAT) gene, named NtAAT-BC and NtAAT-MD. amino acid homology between the two loci reached 98%. High conservative belong to, and AAT amino acid species homology is relatively low, NtAAT-BC and NtAAT-MD contained AAT gene among species are ubiquitous and highly conserved motifs HXXXD and FGWG, can be the beginning of Step two has been successfully determined from Narcissus isolated AAT.6. gene by real-time fluorescence quantitative PCR analysis of two kinds of Narcissus in the opening process of alcohol acyltransferase (AAT) gene expression patterns, the results show that the different stages of AAT gene in Narcissus varieties and flower open exists expression pattern and activity of AAT differential expression of AAT gene. The bride 'crown, the same trend of ester compounds, may affect the ocean water Xian among different varieties of esters synthesis synthesis regulation of.AAT gene expression between' bride 'AAT' crown gene is likely the bride 'crown esters to participate.

【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S682.21

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