發(fā)布時(shí)間：2018-04-28 02:20

  本文選題：顛茄 + 茉莉酸甲酯��； 參考：《西南大學(xué)》2017年碩士論文

【摘要】：顛茄(Atropa belladonna L.)是茄科顛茄屬的重要藥用植物,主要次生代謝產(chǎn)物莨菪堿與東莨菪堿作為重要的抗膽堿類藥物廣泛應(yīng)用,但由于在藥用植物中的含量低,其產(chǎn)量遠(yuǎn)遠(yuǎn)低于市場(chǎng)需求。而用外源添加物處理植物是常用且有效的方法,茉莉酸甲酯(MeJA)作為一種對(duì)植物生長(zhǎng)具有廣泛調(diào)節(jié)作用的物質(zhì),已研究發(fā)現(xiàn)能夠提高多種藥用植物中的次生代謝物含量。本試驗(yàn)以顛茄為材料,設(shè)置了不同濃度的MeJA處理組,0μmol·L-1、100μmol·L-1、200μmol·L-1、300μmol·L-1、400μmol·L-1,研究不同濃度MeJA對(duì)顛茄植株生長(zhǎng)、生理特性、葉片中主要生物堿含量的影響,并從氮代謝、托品烷類生物堿(TAs)生物合成途徑中的關(guān)鍵中間產(chǎn)物以及關(guān)鍵酶基因表達(dá)量等方面,展開對(duì)MeJA調(diào)控顛茄TAs的作用機(jī)理研究。主要研究結(jié)論如下:1.比較不同濃度MeJA處理對(duì)顛茄生理特性的影響,結(jié)果表明:MeJA處理后顛茄葉片中葉綠素a與葉綠素b含量均顯著降低,說(shuō)明MeJA能夠促進(jìn)顛茄葉綠素降解或抑制葉綠素的合成。較低濃度的MeJA(100μmol·L-1、200μmol·L-1)處理在整個(gè)處理期都能顯著提高顛茄葉片中脯氨酸含量,較高濃度MeJA(300μmol·L-1、400μmol·L-1)隨處理時(shí)間的延長(zhǎng)呈先升高后降低或平衡的趨勢(shì),說(shuō)明MeJA可以提高顛茄葉片脯氨酸含量,降低其滲透勢(shì),保持水壓,減少對(duì)細(xì)胞的傷害。MeJA處理顯著提高了顛茄葉片中可溶性蛋白含量,可能是MeJA刺激了顛茄葉片中防御蛋白的合成,防御系統(tǒng)啟動(dòng)時(shí)各種催化酶類大量合成,從而影響顛茄的生理生長(zhǎng)代謝等。2.研究MeJA對(duì)顛茄生長(zhǎng)與主要生物堿含量的影響發(fā)現(xiàn):MeJA處理能夠抑制顛茄的生長(zhǎng),且隨處理時(shí)間的延長(zhǎng)抑制作用越明顯。顛茄葉片中莨菪堿與東莨菪堿含量在MeJA處理后變化趨勢(shì)基本一致,100μmol·L-1MeJA要處理14 d后才能顯著提高主要生物堿含量,而200μmol·L-1、300μmol·L-1、400μmol·L-1 MeJA處理7 d就能顯著提高顛茄葉片兩種生物堿的含量。其中200μmol·L-1在處理28 d時(shí),莨菪堿與東莨菪堿含量達(dá)到最大值,分別為對(duì)照的1.706倍與1.807倍�？傮w來(lái)看,茉莉酸甲酯能促進(jìn)顛茄莨菪堿、東莨菪堿的合成。3.為闡明MeJA調(diào)控顛茄葉片中莨菪堿與東莨菪堿的機(jī)理,對(duì)處理28 d時(shí)顛茄葉片中NO、氮代謝關(guān)鍵酶、TAs生物合成途徑精氨酸脫羧酶(ADC)與鳥氨酸脫羧酶(ODC)、多胺含量以及葉片和根中關(guān)鍵酶基因表達(dá)量進(jìn)行研究。研究發(fā)現(xiàn):MeJA可刺激顛茄葉片中NO的迸發(fā),但變化趨勢(shì)不同于主要生物堿含量。MeJA處理可以降低硝態(tài)氮含量,隨處理濃度升高,硝酸還原酶(NR)與谷氨酰胺合成酶(GS)活性變化趨勢(shì)一致,NR活性得到提高,GS活性除400μmol·L-1處理也得到提高,總體來(lái)看MeJA提高了氮代謝水平。MeJA還可提高ADC活性以及200μmol·L-1、300μmol·L-1處理時(shí)ODC的活性。腐胺含量在MeJA處理后顯著提高,而精胺與亞精胺變化較小。但主要生物堿含量與腐胺的含量變化不完全一致,可能還與TAs合成途徑下游關(guān)鍵酶有關(guān)。除100μmol·L-1處理外,MeJA對(duì)顛茄根中pmt的表達(dá)無(wú)顯著影響。但促進(jìn)了葉中trⅠ以及根中trⅠ和h6h的表達(dá),根中trⅠ表達(dá)量的變化趨勢(shì)與莨菪堿和東莨菪堿含量的變化基本一致,能夠促進(jìn)代謝向合成莨菪堿和東莨菪堿的方向移動(dòng),h6h可促進(jìn)莨菪堿生成東莨菪堿。綜上,推測(cè)MeJA對(duì)顛茄托品烷類生物堿含量影響的機(jī)理,主要是通過(guò)促進(jìn)顛茄根中trⅠ和h6h基因的表達(dá),促進(jìn)次生代謝向合成莨菪堿和東莨菪堿的方向進(jìn)行。此外,適宜濃度的MeJA還可提高顛茄葉片中氮代謝水平,提高ADC、ODC酶活性以及腐胺含量,從而為下游TAs的合成提供充足的前體物質(zhì)。
[Abstract]:Atropa belladonna L. (belladonna) is an important medicinal plant of the Solanaceae. The main secondary metabolites, scopolamine and scopolamine, are widely used as important anticholinergic drugs. However, the yield is far below the market demand because of the low content of the medicinal plants, and the treatment of plants with external additives is a common and effective method. Methyl jasmonate (MeJA), as a substance which has a wide regulating effect on plant growth, has been found to be able to increase the secondary metabolite content in a variety of medicinal plants. This experiment uses belladonna to set up different concentrations of MeJA treatment group, 0 u mol. L-1100, L-1200, L-1300, L-1300, L-1300, L-1400, mol. The effects of MeJA on the growth and physiological characteristics of belladonna, the main alkaloid content in the leaves and the key intermediate products in the biosynthesis pathway of the TAs biosynthesis pathway and the gene expression of the key enzymes were studied. The main conclusions were as follows: 1. the main conclusions are as follows: 1. The effects of the same concentration of MeJA on the physiological characteristics of belladonna showed that the content of chlorophyll a and chlorophyll b in the leaves of belladonna decreased significantly after MeJA treatment, indicating that MeJA could promote the degradation of belladonna chlorophyll or inhibit the synthesis of chlorophyll. The lower concentration MeJA (100 mu mol. L-1200, mol L-1) can be significantly improved in the whole treatment period. Proline content in belladonna leaves, high concentration of MeJA (300 mu mol. L-1400 Mu mol. L-1) increased first and then decreased or balanced with the prolongation of treatment time, indicating that MeJA could improve the proline content of belladonna leaves, decrease its osmotic potential, maintain water pressure, and reduce the damage to cells by.MeJA treatment, which significantly improved the soluble eggs in belladonna leaves. The white content may be that MeJA stimulates the synthesis of defensive proteins in the belladonna leaves, and a large number of catalytic enzymes are synthesized at the start of the defense system, which affects the physiological growth and metabolism of the belladonna and other.2. studies on the effects of MeJA on the growth of Belladonna and the main alkaloid content. It is found that MeJA treatment can inhibit the growth of Belladonna and inhibit the growth of belladonna. The more obvious the production is. The changes in the contents of scopolamine and scopolamine in the vane of belladonna are basically the same after MeJA treatment. The content of the main alkaloids can be significantly increased after 100 mu mol. L-1MeJA is treated with 14 d. The content of two alkaloids of the high belladonna leaves can be significantly raised by 7 d. 20, mol. L-1400, mol L-1 MeJA. 0 Mu mol. L-1 reached the maximum value of scopolamine and scopolamine at the maximum value of 28 d, 1.706 times and 1.807 times the control, respectively. In general, methyl jasmonate could promote Belladonna and scopolamine synthesis.3. to clarify the mechanism of MeJA regulation of belladonna scopolamine and scopolamine, NO, nitrogen generation in belladonna leaves at 28 d treatment. Key enzyme, TAs biosynthesis pathway, arginine decarboxylase (ADC) and ornithine decarboxylase (ODC), polyamine content and gene expression of key enzymes in leaves and roots. The study found that MeJA can stimulate the burst of NO in belladonna leaves, but the change trend is different from the main alkaloid content.MeJA treatment can reduce nitrate nitrogen content everywhere. As the concentration increased, the activity of nitrate reductase (NR) and glutamine synthetase (GS) was the same, the activity of NR was improved and the activity of GS was improved in addition to 400 mu mol. L-1. In general, the MeJA increased the nitrogen metabolism level and the activity of ADC activity, as well as the activity of 200 mu mol. The changes of spermine and spermine were smaller, but the content of the main alkaloids was not exactly consistent with the changes in the content of the putrescine. It may also be related to the key enzymes in the downstream of TAs synthesis. Except for the treatment of 100 mu mol. L-1, MeJA has no significant influence on the expression of PMT in the belladonna root, but it promotes the expression of TR I and h6h in the leaves of TR I and the roots. The change in the expression of TR I was basically consistent with the changes in the content of scopolamine and scopolamine, which could promote the direction of metabolism to the direction of scopolamine and scopolamine. H6h could promote the formation of scopolamine. The mechanism of the effect of MeJA on the content of alkanoid alkaloids in belladonna, mainly by promoting the TR in belladonna root. The expression of I and h6h gene promotes the direction of secondary metabolism to synthesis of scopolamine and scopolamine. In addition, the suitable concentration of MeJA can also improve the level of nitrogen metabolism in the vane of belladonna, improve the activity of ADC, ODC enzyme and the content of the putrescine, thus providing sufficient precursors for the synthesis of TAs in the lower reaches.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：Q946

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 強(qiáng)瑋;侯艷玲;李笑;夏科;廖志華;;木本曼陀羅中催化東莨菪堿生物合成關(guān)鍵步驟的H6H基因克隆與表達(dá)分析[J];藥學(xué)學(xué)報(bào);2015年10期

2 張梅;林棋;婁本勇;潘大仁;周以飛;;茉莉酸甲酯對(duì)錦繡杜鵑黃酮的累積及相關(guān)酶活性的影響[J];甘肅農(nóng)業(yè)大學(xué)學(xué)報(bào);2015年04期

3 湯紹虎;胡俠;;外源茉莉酸甲酯對(duì)受傷苦瓜幼苗生理特性的影響[J];西南大學(xué)學(xué)報(bào)(自然科學(xué)版);2014年08期

4 行冰玉;黨小琳;張婧一;汪波;陳子逸;董娟娥;;茉莉酸甲酯對(duì)丹參培養(yǎng)細(xì)胞中迷迭香酸生物合成及相關(guān)酶活性的影響[J];植物生理學(xué)報(bào);2013年12期

5 謝陽(yáng)姣;何志鵬;林偉;;茉莉酸甲酯對(duì)苦玄參生長(zhǎng)及苦玄參苷積累的影響研究[J];作物雜志;2013年02期

6 盧衍;王細(xì)榮;陳敏;廖志華;;托品烷類生物堿生物合成分子生物學(xué)與代謝工程[J];中草藥;2012年05期

7 張智慧;聶燕芳;何磊;高元媛;李云鋒;王振中;;外源茉莉酸甲酯誘導(dǎo)的水稻葉片蛋白質(zhì)差異表達(dá)分析[J];植物病理學(xué)報(bào);2012年02期

8 李文淵;高偉;趙靜;崔光紅;邵愛(ài)娟;黃璐琦;;基于茉莉酸甲酯誘導(dǎo)的丹參毛狀根酚酸類成分次生代謝機(jī)制研究[J];中國(guó)中藥雜志;2012年01期

9 鄧朝暉;羅充;劉彬;陳玲;譚金玉;;曼陀羅藥用價(jià)值的開發(fā)和利用[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2011年07期

10 段小華;鄧澤元;賓金華;;茉莉酸甲酯對(duì)水稻幼苗抗冷性的影響[J];植物生理學(xué)通訊;2009年09期

相關(guān)博士學(xué)位論文 前2條

1 胡永建;蟲害馬尾松鄰株信號(hào)通訊及茉莉酸甲酯誘導(dǎo)的蛋白表達(dá)差異研究[D];北京林業(yè)大學(xué);2010年

2 李鳳嵐;紅豆杉紫杉烷13α-，，14β-羥基化酶基因調(diào)控及茉莉酸甲酯誘導(dǎo)下的蛋白表達(dá)[D];北京協(xié)和醫(yī)學(xué)院;2009年

相關(guān)碩士學(xué)位論文 前10條

1 王中平;顛茄精氨酸脫羧酶基因的克隆與功能分析[D];西南大學(xué);2016年

2 劉佳;誘導(dǎo)子對(duì)顛茄毛狀根生長(zhǎng)及托品烷類生物堿代謝機(jī)理初步研究[D];西南大學(xué);2016年

3 李琳琳;茉莉酸甲酯對(duì)顛茄毛狀根的生長(zhǎng)及次生代謝產(chǎn)物產(chǎn)生的影響[D];西南大學(xué);2015年

4 孫際薇;茉莉酸甲酯對(duì)曼陀羅毛狀根的生長(zhǎng)及次生代謝產(chǎn)物產(chǎn)生的影響[D];西南大學(xué);2014年

5 秦白富;顛茄ODC基因克隆和功能分析[D];西南大學(xué);2014年

6 王亞雄;顛茄類托品酮還原酶基因克隆及功能研究[D];西南大學(xué);2014年

7 施江;外源茉莉酸甲酯誘導(dǎo)對(duì)茶樹鮮葉次生代謝產(chǎn)物的影響[D];中國(guó)農(nóng)業(yè)科學(xué)院;2014年

8 強(qiáng)瑋;顛茄TRI基因的克隆、功能驗(yàn)證及超量表達(dá)對(duì)顛茄托品烷生物堿合成的影響[D];西南大學(xué);2012年

9 李春曉;MeJA和SA對(duì)白樺幼樹三萜合成調(diào)控及FPS基因克隆[D];東北林業(yè)大學(xué);2012年

10 龔小貴;不同濃度茉莉酸甲酯對(duì)馬藍(lán)生理與蛋白質(zhì)的影響[D];福建農(nóng)林大學(xué);2011年

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