本文關鍵詞：中間錦雞兒肉桂酰輔酶A還原酶基因的克隆和功能鑒定　出處：《內(nèi)蒙古農(nóng)業(yè)大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 中間錦雞兒 肉桂酰輔酶A還原酶 木質(zhì)素 克隆

【摘要】：木質(zhì)素是一種芳香雜聚物,它是自然界中在總量上僅次于纖維素的天然有機高分子物質(zhì),已得到廣泛的研究。本研究從中間錦雞兒中克隆得到兩個木質(zhì)素合成途徑上的肉桂酰輔酶A還原酶基因,分別命名為CiCCR2和CiCCR3,并構建了植物表達載體,轉(zhuǎn)化野生型擬南芥并獲得純合體株系,具體結果如下:(1)中間錦雞兒CiCCR2基因開放閱讀框長為897bp,gDNA共有4個內(nèi)含子,分別長131bp、1351bp、176bp、222bp;編碼299個氨基酸,推測的分子量為33.696KD,等電點為5.11,平均疏水指數(shù)(GRAVY)為-0.288;CiCCR3基因開放閱讀框為966bp,gDNA 有 5 個內(nèi)含子,分別長 829bp、856bp、197bp、522bp、1191bp,編碼 322 個氨基酸,推測的分子量為35.91KD,等電點為6.01,平均疏水指數(shù)為-0.173。(2)構建過表達載體p35S::CiCCR2和p35S::CiCCR3,并轉(zhuǎn)化野生型擬南芥Columbia-0,分別得到純合體8個株系、9個株系。(3)過表達CiCCR2和CiCCR3基因的擬南芥,在成熟時期和幼苗時期木質(zhì)素含量均高于野生型擬南芥組織化學染色也驗證了轉(zhuǎn)基因株系木質(zhì)素積累較野生型擬南芥多。并且CiCCR2和CiCCR3過表達植株的鮮重和干重比野生型增加。
[Abstract]:Lignin is a kind of aromatic heterozygous. It is a natural organic polymer in the natural world, second only to cellulose, and has been widely studied. The cloning of cinnamoyl coenzyme A reductase gene two lignin biosynthesis pathway from Caragana intermedia, named CiCCR2 and CiCCR3, and constructed the plant expression vector and transformed into wild type Arabidopsis and obtain homozygous lines, the specific results are as follows: (1) Caragana CiCCR2 gene open reading frame 897bp long, gDNA has 4 introns, respectively 131bp, 1351bp, 176bp, long 222bp; encoding 299 amino acids. The predicted molecular weight of 33.696KD and isoelectric point is 5.11, the average hydrophobicity index (GRAVY) for -0.288; on the open reading frame of 966bp CiCCR3 gene has 5 introns, gDNA respectively, 829bp, 856bp, long 197bp, 522bp, 1191bp, encoding 322 amino acids. The predicted molecular weight of 35.91KD and isoelectric point is 6.01, the average hydrophobicity index was -0.173. (2) the over expression vector p35S:: CiCCR2 and p35S:: CiCCR3 were constructed and transformed into wild Arabidopsis Columbia-0, and 8 strains of homozygous lines and 9 strains were obtained. (3) the overexpression of CiCCR2 and CiCCR3 genes in Arabidopsis thaliana showed higher lignin content in mature and seedling stage than in wild Arabidopsis thaliana. Histochemical staining also confirmed that the accumulation of lignin in transgenic plants was more than that in wild Arabidopsis thaliana. The fresh weight and dry weight of the overexpressed plants of CiCCR2 and CiCCR3 increased as compared with the wild type.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q943.2

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