本文選題：絲瓜 + 酚類物質(zhì)　； 參考：《福建農(nóng)林大學(xué)》2017年碩士論文

【摘要】：絲瓜藥食兩用,以其柔嫩多汁的果實深受人們的喜愛,在中國各地普遍種植。絲瓜果肉易出現(xiàn)褐變,影響其商品價值,揭示絲瓜褐變機理成為絲瓜育種和采后的研究重點。酚類物質(zhì)與果蔬的褐變密切相關(guān),是果蔬酶促褐變的主要條件之一。為探究引起絲瓜酶促褐變主要酚類物質(zhì)的組分,從中篩選出抗褐變品種,本文以11個絲瓜品種為試材,優(yōu)化了絲瓜酚類物質(zhì)的提取方法,建立了絲瓜超高效液相色譜快速檢測體系,分析了絲瓜酚類物質(zhì)組分及其含量。通過體外模擬酚類物質(zhì)的酶促褐變過程,結(jié)合超高效液相色譜法,確定了引起酶促褐變的主要酚類組分,并篩選出耐褐變絲瓜品種。對絲瓜酚類化合物生物合成中的關(guān)鍵酶LcPAL1、LcPAL2基因表達(dá)模式進(jìn)行了分析。本研究為絲瓜抗褐變育種提供理論依據(jù)與方法。主要結(jié)果如下:1.優(yōu)化了絲瓜酚類物質(zhì)提取方法,結(jié)果顯示:以乙醇為提取劑,料液比1g:20mL,強酸(pH3～4)環(huán)境下進(jìn)行冰浴超聲粉碎,絲瓜酚類物質(zhì)提取效果最好。2.建立了超高效液相色譜法分析絲瓜酚類物質(zhì)組分及其含量,結(jié)果如下:液相色譜柱為 ACQUITYUPLCBEHC18(2.1mm×100mm,1.7 μm),流動相為甲醇(A)/0.3%乙酸水溶液(B),柱溫32℃,進(jìn)樣量2 μL,流速為0.25 mL·min-1。采用梯度洗脫,洗脫程序0～0.5 min,95%B;0.5～17.5min,95%～75%B;17.5～18min,75%～95%B。檢測波長選取283nm。14種酚類化合物標(biāo)準(zhǔn)品在18 min內(nèi)完全分離,樣品中檢測出14種酚酸。線性范圍在0.05～4.0 mg· L-1,檢出限(S/N=3)為0.001～0.049mg·L-1,決定系數(shù)均大于0.999,精密度、重復(fù)性、穩(wěn)定性相對標(biāo)準(zhǔn)偏差均小于5%。平均回收率在95.36%～105.58%,相對標(biāo)準(zhǔn)偏差在 0.80%～3.64%。3.通過絲瓜酶促褐變體外模擬反應(yīng),確定了引起絲瓜酶促褐變的酚類物質(zhì)為:綠原酸、對羥基苯甲酸和4-甲基兒茶酚,其中綠原酸為主要褐變底物。根據(jù)褐變底物的含量篩選出了耐褐變絲瓜品種C1、C7 和 C9。4.對絲瓜酚類化合物生物合成中的關(guān)鍵酶LcPAL1、LcPAL2基因進(jìn)行表達(dá)分析,結(jié)果表明,2個基因都具有組織表達(dá)特異性,在葉中的表達(dá)量最高,根中的表達(dá)量也顯著高于莖、花和果,LcPAL1基因的表達(dá)順序為葉根果花莖,LcPAL2基因的表達(dá)順序為葉根莖花果。絲瓜LcPAL1、LcPAL2基因在不同品種間呈差異表達(dá),其中有棱絲瓜的表達(dá)量顯著低于肉絲瓜。絲瓜果肉在25℃鮮切處理和4℃低溫冷藏過程中LcPAL1、LcPAL2基因均表現(xiàn)為先上升再下降的表達(dá)趨勢,結(jié)合所測PAL酶活和總酚含量相關(guān)性,推測絲瓜LcPAL1、LcPAL 基因在鮮切和冷藏過程中會調(diào)控苯丙氨酸解氨酶合成酚類物質(zhì),促進(jìn)絲瓜酶促褐變的發(fā)生。
[Abstract]:The loofah is popular with its tender and juicy fruit. It is widely cultivated in all parts of China. It is easily browning and affects its commodity value. It is revealed that the browning mechanism of the loofah has become the focus of the research on the breeding and post harvest of the silk gourd. The phenolic substances are closely related to the browning of fruits and vegetables, and it is one of the main conditions for the enzymatic browning of fruits and vegetables. In order to explore the components of the main phenolic substances that cause the browning of the loofah, the anti browning varieties were screened. In this paper, 11 varieties of Luffa were used as the test materials to optimize the extraction method of the phenolic compounds of the Luffa. The rapid detection system for the liquid chromatography of the Luffa high performance liquid chromatography was established, and the components and content of the phenolic compounds were analyzed. The enzymatic browning process, combined with super high performance liquid chromatography, was used to determine the main phenolic components that cause enzymatic browning, and to screen out browning gourd varieties. The key enzyme LcPAL1 and LcPAL2 gene expression patterns in the biosynthesis of Luffa phenolic compounds were analyzed. This study provides a theoretical basis and method for the anti Browning breeding of silk gourd. The main results are as follows: 1. the extraction method of Luffa phenols was optimized. The results showed that using ethanol as the extraction agent, the material and liquid ratio 1g:20mL, strong acid (pH3 ~ 4) under the environment of ice bath ultrasonic pulverization, the extraction effect of Luffa phenolic substances was best.2. established by ultra high performance liquid chromatography to analyze the composition and content of silk gourd substances. The results are as follows: liquid phase color The spectrum column is ACQUITYUPLCBEHC18 (2.1mm x 100mm, 1.7 mu m), the flow phase is methanol (A) /0.3% acetic acid water solution (B), the column temperature is 32 c, the sample volume is 2 u L, the flow rate is 0.25 mL min-1., the gradient elution is used, the elution program is 0 ~ 0.5 min, 95%B, 0.5 to 95% ~ 18 min was completely separated and 14 kinds of phenolic acids were detected. The linear range was from 0.05 to 4 mg. L-1, the detection limit (S/N=3) was 0.001 to 0.049mg. L-1, the determination coefficient was greater than 0.999, the precision, repeatability, the relative standard deviation of stability were less than the average recovery of 5%. in 95.36% to 105.58%, and the relative standard deviation was 0.80% to 3.64%.3. through the silk gourd. Enzymatic browning in vitro simulated reaction to determine that the phenolic substances that cause the browning of the loofa are chlorogenic acid, para hydroxy benzoic acid and 4- methyl catechol, in which chlorogenic acid is the main browning substrate. According to the content of browning substrates, C1, C7 and C9.4. are the key enzymes in the biosynthesis of Luffa phenolic compounds, LcPA L1, LcPAL2 gene expression analysis, the results show that the 2 genes all have the specificity of tissue expression, the highest expression in the leaves, the amount of expression in the root is significantly higher than the stem, flower and fruit, the LcPAL1 gene expression sequence is the leaf root fruit stem, the LcPAL2 gene expression sequence is Ye Genjing flower and fruit. The LcPAL1, LcPAL2 gene of the silk gourd is in the different varieties. The expression of gourd was significantly lower than that of meat gourd. The LcPAL1, LcPAL2 gene of the pulp at 25 centigrade and 4 degrees centigrade in cold storage, showed a tendency to increase first and then decrease, and combined with the correlation between the PAL enzyme activity and the total phenol content, the LcPAL1 of the loofah was conjectured and the LcPAL gene was in the process of fresh cut and cold storage. It can regulate phenylalanine ammonia lyase to synthesize phenols, and promote the occurrence of enzymatic browning of Luffa.

【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】：S642.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王宇;張鍇;孫偉明;王帥;曹紅梅;董秋平;張華雪;李凱;;冀東野生大豆對大豆花葉病毒的抗性鑒定及抗病反應(yīng)[J];大豆科學(xué);2017年01期

2 田華;朱艷梅;董瑜;劉偉;孔凡玉;王靜;;毀滅炭疽菌Colletotrichum destructivum誘抗蛋白對煙草抗病性的誘導(dǎo)[J];植物保護(hù)學(xué)報;2017年01期

3 楊宜紅;陸燕;吳穎靜;馮自力;唐燦明;;棉花品種黃萎病抗性與免疫反應(yīng)的關(guān)系[J];核農(nóng)學(xué)報;2017年03期

4 權(quán)伍榮;康優(yōu);沈明浩;;蘋果梨中多酚氧化酶的性質(zhì)及多酚成分定性[J];食品研究與開發(fā);2016年18期

5 張庭廷;韓玉珍;何宗祥;汪好芬;;酚酸類物質(zhì)對銅綠微囊藻以及蛋白核小球藻的抑藻作用[J];衛(wèi)生研究;2016年03期

6 王傳義;呂國新;朱啟法;徐秀紅;蔡憲杰;解燕;王朝佐;任杰;趙華武;;烤煙烘烤特性與煙草多酚氧化酶活性相關(guān)性研究[J];湖北農(nóng)業(yè)科學(xué);2016年06期

7 李大忠;朱海生;康玉妹;溫慶放;李永平;康建坂;;普通絲瓜新品種‘福研1號’選育[J];亞熱帶植物科學(xué);2016年01期

8 劉遙;謝揚;祁健斌;田立文;;過山楓中總芪類的提取和純化[J];湖北農(nóng)業(yè)科學(xué);2016年03期

9 劉曉宇;傅海燕;黃國和;柴天;高攀峰;吳義誠;;美人蕉有機酸組分對銅綠微囊藻的化感作用[J];環(huán)境工程學(xué)報;2015年12期

10 黃樹蘋;徐長城;談杰;張敏;王春麗;陳霞;甘莉;談太明;;不同影響因素下普通絲瓜果肉褐變度、多酚氧化酶活性及多酚質(zhì)量比變化研究[J];西南大學(xué)學(xué)報(自然科學(xué)版);2015年09期

相關(guān)會議論文 前1條

1 楊梅;黃曉露;;桉樹連作對土壤多酚氧化酶活性及酚類物質(zhì)含量的影響[A];中國第五屆植物化感作用學(xué)術(shù)研討會論文摘要集[C];2011年

相關(guān)博士學(xué)位論文 前2條

1 吳夏;五味子鮮果多酚提取鑒定及其對受損HepG2細(xì)胞保護(hù)作用的研究[D];中國農(nóng)業(yè)大學(xué);2014年

2 曲留柱;香蕉多酚氧化酶特性及其催化褐變防控的研究[D];北京林業(yè)大學(xué);2014年

相關(guān)碩士學(xué)位論文 前10條

1 莊尹宏;絲瓜褐變分級與總酚及相關(guān)酶活性分析[D];福建農(nóng)林大學(xué);2016年

2 馮發(fā)進(jìn);垂絲海棠活性成分研究[D];貴州大學(xué);2015年

3 屈政;引發(fā)金銀花褐變的酶類和金銀花中多酚氧化酶提取純化的研究[D];河南科技大學(xué);2015年

4 李博;采后荔枝果皮褐變與POD基因表達(dá)關(guān)系的研究[D];海南大學(xué);2014年

5 陳東明;超聲提取玫瑰花渣中多酚、多糖的工藝研究[D];鄭州大學(xué);2014年

6 鄒麗紅;梨果肉酶促褐變機理研究[D];河北農(nóng)業(yè)大學(xué);2012年

7 周艷瓊;茉莉酸甲酯對楊樹誘導(dǎo)抗性的研究[D];南京林業(yè)大學(xué);2011年

8 張寬朝;霍山石斛苯丙氨酸解氨酶的分離純化、酶學(xué)特性及外界影響因素研究[D];安徽農(nóng)業(yè)大學(xué);2009年

9 張永麗;荔枝果皮褐變過程中POD活性變化及其基因表達(dá)分析[D];華南熱帶農(nóng)業(yè)大學(xué);2007年

10 劉衛(wèi)紅;銀杏葉苯丙氨酸解氨酶特性及其對葉黃酮含量調(diào)控的研究[D];華中農(nóng)業(yè)大學(xué);2003年

，

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