亞麻BRs相關(guān)基因的克隆與功能初步驗證
發(fā)布時間:2018-05-06 00:28
本文選題:亞麻 + LuDWF4; 參考:《新疆大學(xué)》2017年碩士論文
【摘要】:油菜素甾醇(BRs)在調(diào)節(jié)植物細胞伸長,細胞壁纖維素合成等發(fā)育過程中起重要作用。亞麻(Linum usitatissimum L.)作為紡織、工業(yè)、藥用原料,經(jīng)濟價值極高。亞麻株高和纖維長度決定纖維產(chǎn)量和品質(zhì)。本研究以亞麻品種“范妮”為材料,通過同源序列克隆法從亞麻葉片中獲得BR生物合成途徑中的限速酶LuDWF4,BR信號轉(zhuǎn)導(dǎo)途徑中的受體蛋白和轉(zhuǎn)錄因子Lu BRI1和LuBES1基因全序列,在野生型擬南芥Col-0和BRs受體弱突變體bril-301中進行初步功能驗證。同時,探討了亞麻快速生長期與細胞壁形成相關(guān)基因的表達,為亞麻纖維發(fā)育研究奠定理論基礎(chǔ)。主要結(jié)論如下:(1)通過同源序列克隆法從亞麻葉片中克隆了LuDWF4、LuBRI1、cLuBES1和gLuBES1基因,全長分別為1479 bp、3579 bp、978 bp和1428 bp。(2)成功構(gòu)建植物表達載體,分別獲得了4個(D_2,D_9,D_(10),D_(16))、3個(BR_3,BR_4,BR_(12))和3個(BE_1,BE_4,BE_(11))株系的LuDWF4、LuBRI1和gLuBES1單拷貝插入轉(zhuǎn)基因植株。(3)過表達LuDWF4基因促進擬南芥Col-0植株增大、花期提前、花梗增高;過表達LuBRI1基因或gLuBES1基因促進bril-301植株的生長發(fā)育,挽救了bril-301矮化表型。光照和黑暗條件下,外施epi BL促進Col-0、bril-301及LuBRI1和gLuBES1轉(zhuǎn)基因株系下胚軸伸長。(4)快速生長期亞麻莖韌皮纖維細胞厚度TOPMIDBOT區(qū),SP點下部細胞壁沒有次生加厚過程;LuBGAL3、LuBGAL5、LuBGAL6、LuBGAL9、LuCESA1、LuCESA3、LuCESA9和LuCESA10在亞麻細胞壁細胞伸長過程中起作用;LuBGAL1主要促進亞麻細胞壁加厚過程;LuSuSy和LuXTH4亞麻細胞壁發(fā)育中發(fā)揮作用。
[Abstract]:Brassin sterol (BRs) plays an important role in the regulation of plant cell elongation and cell wall cellulose synthesis. Linum usitatissimum L. As textile, industrial, medicinal raw materials, economic value is very high. Flax plant height and fiber length determine fiber yield and quality. In this study, Fanny, a flax variety, was used to obtain the receptor protein and the complete sequence of Lu BRI1 and LuBES1 genes in Br biosynthesis pathway from flax leaves by homologous sequence cloning. The function of wild-type Arabidopsis thaliana Col-0 and BRs acceptor weak mutants bril-301 was preliminarily verified. At the same time, the expression of genes related to cell wall formation in the fast growing stage of flax was discussed, which laid a theoretical foundation for the study of flax fiber development. The main conclusions are as follows: (1) the LuDWF4 (LuBRI1) cLuBES1 and gLuBES1 genes were cloned from flax leaves by homologous sequence cloning. The total length of LuDWF4 and LuBRI1 gene were 1479 BP, 3579 BP, 978 BP and 1428 BP, respectively) and the plant expression vectors were successfully constructed. Four D2D9 / D10 / D _ (10) / D _ _ _ / _ _ _ Overexpression of LuBRI1 gene or gLuBES1 gene promoted the growth and development of bril-301 plants and saved bril-301 dwarfing phenotype. In light and darkness, Application of epi BL promoted elongation of Hypocotyl of Col-0 Bril-301 and LuBRI1 and gLuBES1 transgenic Lines. 4) during the Rapid growing period, there was no secondary thickening process of the cell wall at the SP site of the thickness TOPMIDBOT zone of flax stem phloem fiber cells. There was no secondary thickening process of LuBGAL3, LuBGAL5, LuBGAL9, LuCESA1, LuCESA3, LuCESA9 and LuCESA10 in the cell wall of flax. LuBGAL1 plays an important role in the development of cell wall of flax, LuSuSy and LuXTH4.
【學(xué)位授予單位】:新疆大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:Q943.2;S563.2
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2 李懷玖;喻身啟;邱榮;;推廣的BRS變換下W—T恒等式和SU(2)規(guī)范場中ξ_c~b的一種表示[J];遼寧師范大學(xué)學(xué)報(自然科學(xué)版);1983年03期
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