【摘要】：在香蕉基因組中尋找具有抗倒伏功能的基因,為培育抗倒伏香蕉新品種奠定基礎(chǔ)。香豆酸:Co A連接酶(4CL)是植物木質(zhì)素合成的限速酶。香蕉A基因組中有25個(gè)4CL基因同源序列。高粱基因bmr2是一個(gè)已經(jīng)被鑒定的具有4CL功能的基因。與bmr2進(jìn)行序列比對(duì),將候選基因在大腸桿菌中過(guò)量表達(dá),提取重組蛋白,研究其催化活性。將候選基因在煙草中過(guò)量表達(dá),研究轉(zhuǎn)基因植株與對(duì)照的木質(zhì)素含量和莖稈物理強(qiáng)度。香蕉4-香豆酸:Co A連接酶基因Mu4CL15預(yù)測(cè)的底物結(jié)合域序列、二級(jí)結(jié)構(gòu)、三級(jí)結(jié)構(gòu)都與bmr2的相應(yīng)序列具有高度的相似性。將Mu4CL15在大腸桿菌中表達(dá)并提取純化,重組蛋白質(zhì)催化效率最高的底物是4-香豆酸,其次是咖啡酸、阿魏酸和芥子酸。將Mu4CL15在煙草中進(jìn)行轉(zhuǎn)化后發(fā)現(xiàn),轉(zhuǎn)基因植株莖稈的木質(zhì)素含量顯著高于只轉(zhuǎn)化空載體的野生型。轉(zhuǎn)基因幼苗的莖稈強(qiáng)度顯著高于只轉(zhuǎn)化空載體的野生型。在煙草中過(guò)量表達(dá)Mu4CL15能夠增加煙草植株莖稈的木質(zhì)素含量和抗倒伏能力。通過(guò)過(guò)量表達(dá)基因Mu4CL15極有可能能夠提高香蕉植株假莖的抗倒伏能力。
[Abstract]:To find genes with lodging resistance in banana genome, which laid a foundation for the cultivation of new lodging resistant banana varieties. Coumaric acid: Co A ligase (4CL) is a rate-limiting enzyme for plant lignin synthesis. There are 25 homologous sequences of 4CL gene in banana A genome. Sorghum gene bmr2 is a gene with 4CL function that has been identified. Compared with bmr2, the candidate genes were overexpressed in E. coli, the recombinant protein was extracted, and its catalytic activity was studied. The lignin content and stem physical strength of transgenic plants and controls were studied by overexpression of candidate genes in tobacco. The substrate binding domain sequence, secondary structure and tertiary structure predicted by banana 4-coumaric acid: Co A ligase gene Mu4CL15 are highly similar to the corresponding sequences of bmr2. Mu4CL15 was expressed in E. coli and purified. The substrate with the highest catalytic efficiency of recombinant protein was 4-coumaric acid, followed by caffeine, ferulic acid and erucic acid. After transformation of Mu4CL15 in tobacco, it was found that the lignin content of the stem of the transgenic plant was significantly higher than that of the wild type only transformed with empty carrier. The stem strength of transgenic seedlings was significantly higher than that of wild type only transformed into empty carrier. Overexpression of Mu4CL15 in tobacco could increase lignin content and lodging resistance of tobacco stems. Overexpression of gene Mu4CL15 is likely to improve the lodging resistance of false stems of banana plants.
【作者單位】： 中國(guó)熱帶農(nóng)業(yè)科學(xué)院�？趯�(shí)驗(yàn)站;海南省香蕉遺傳改良重點(diǎn)實(shí)驗(yàn)室;海南大學(xué)農(nóng)學(xué)院;
【基金】：海南省重點(diǎn)研發(fā)項(xiàng)目(No.ZDYF2016069) 國(guó)家自然科學(xué)基金(No.31371696) 農(nóng)業(yè)部948項(xiàng)目(No.2015-Z21)共同資助
【分類號(hào)】：Q943.2;S668.1

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