亚麻BRs相关基因的克隆与功能初步验证

发布时间：2018-05-06 00:28

  本文选题：亚麻 + LuDWF4　； 参考：《新疆大学》2017年硕士论文

【摘要】：油菜素甾醇(BRs)在调节植物细胞伸长,细胞壁纤维素合成等发育过程中起重要作用。亚麻(Linum usitatissimum L.)作为纺织、工业、药用原料,经济价值极高。亚麻株高和纤维长度决定纤维产量和品质。本研究以亚麻品种“范妮”为材料,通过同源序列克隆法从亚麻叶片中获得BR生物合成途径中的限速酶LuDWF4,BR信号转导途径中的受体蛋白和转录因子Lu BRI1和LuBES1基因全序列,在野生型拟南芥Col-0和BRs受体弱突变体bril-301中进行初步功能验证。同时,探讨了亚麻快速生长期与细胞壁形成相关基因的表达,为亚麻纤维发育研究奠定理论基础。主要结论如下:(1)通过同源序列克隆法从亚麻叶片中克隆了LuDWF4、LuBRI1、cLuBES1和gLuBES1基因,全长分别为1479 bp、3579 bp、978 bp和1428 bp。(2)成功构建植物表达载体,分别获得了4个(D_2,D_9,D_(10),D_(16))、3个(BR_3,BR_4,BR_(12))和3个(BE_1,BE_4,BE_(11))株系的LuDWF4、LuBRI1和gLuBES1单拷贝插入转基因植株。(3)过表达LuDWF4基因促进拟南芥Col-0植株增大、花期提前、花梗增高;过表达LuBRI1基因或gLuBES1基因促进bril-301植株的生长发育,挽救了bril-301矮化表型。光照和黑暗条件下,外施epi BL促进Col-0、bril-301及LuBRI1和gLuBES1转基因株系下胚轴伸长。(4)快速生长期亚麻茎韧皮纤维细胞厚度TOPMIDBOT区,SP点下部细胞壁没有次生加厚过程;LuBGAL3、LuBGAL5、LuBGAL6、LuBGAL9、LuCESA1、LuCESA3、LuCESA9和LuCESA10在亚麻细胞壁细胞伸长过程中起作用;LuBGAL1主要促进亚麻细胞壁加厚过程;LuSuSy和LuXTH4亚麻细胞壁发育中发挥作用。
[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.
【学位授予单位】：新疆大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2;S563.2

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