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作物学报 ›› 2008, Vol. 34 ›› Issue (02): 337-340.doi: 10.3724/SP.J.1006.2008.00337

• 研究简报 • 上一篇    下一篇

亚麻木质素合成途径中关键酶基因片段的克隆与序列分析

高原1,2;陈信波1,2,*;龙松华1;邓欣1;邹杰2;刘爱玲3   

  1. 1中国农业科学院麻类研究所农业部麻类遗传改良与工程微生物重点实验室, 湖南长沙410205; 2湖南农业大学作物基因工程重点实验室, 湖南长沙410128; 3湖南农业大学生物科学技术学院, 湖南长沙 410128

  • 收稿日期:2007-04-10 修回日期:1900-01-01 出版日期:2008-02-12 网络出版日期:2008-02-12
  • 通讯作者: 陈信波

Cloning and Sequence Analysis of Partial Fragments of Critical Genes Involved in Flax Lignin Synthesis

GAO Yuan12,CHEN Xin-Bo12*,LONG Song-Hua1,DENG Xin1,ZOU Jie2,LIU Ai-Ling3   

  1. 1 Ministry of Agriculture Key Laboratory of Genetic Improvement & Engineering Microbiology for Bast Fiber Crops, Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, Hunan; 2 Crop Gene Engineering Key Laboratory, Hunan Agricultural University, Changsha 410128, Hunan; 3 College of Bio-science and Technology, Hunan Agricultural University, Changsha 410128, Hunan, China

  • Received:2007-04-10 Revised:1900-01-01 Published:2008-02-12 Published online:2008-02-12
  • Contact: CHEN Xin-Bo

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1677

被引次数

15

摘要：

亚麻是一种重要的韧皮纤维作物, 木质素对亚麻纤维的性能和品质具有较大影响。以亚麻(Linum usitatissimum L.)茎秆表皮细胞的mRNA为模板, 利用木质素合成途径中关键酶基因的同源基因保守序列设计简并引物, 通过RT-PCR扩增, 电泳获得13个特异带。分别将这些DNA片段连接到T载体后转化大肠杆菌, 从重组质粒转化菌分别挑取5~8个单菌落测定插入片段序列, 得到关键酶基因新的片段序列8个。生物信息学分析结果表明, 这些新片段分别属于3个基因家族。其中, 2个CCoAOMT基因片段(GenBank登录号为EF214740, EF214741), 3个4CL基因片段(GenBank登录号为EF214737, EF214738, EF214739), 3个F5H基因片段(GenBank登录号为EF214745, EF214746, EF214747), 推测亚麻中这几个木质素代谢关键调控酶基因存在多基因家族。新获得的基因片段序列为进一步克隆全序列和了解调控亚麻木质素的生物合成奠定了基础。

关键词: 亚麻(Linum usitatissimum L.), CCoAOMT基因, 4CL基因, F5H基因, 序列分析

Abstract:

Flax is an important bast fiber crop. High content of lignin in the fiber greatly reduces fiber quality after degumming. The cDNA fragments of the critical genes involved in flax lignin biosynthesis were amplified from the flax bark mRNA by RT-PCR using the degenerate primers designed from conservative regions of existing homologous gene sequences. Thirteen distinct bands were obtained from agarose gel electrophoresis. The fragments from these bands were ligated to T-vector. The recombinant plasmids were transformed into E. coli. Five to eight separate clones transformed from each band were chosen to be sequenced. Sequence analysis and GenBank BLAST revealed eight new sequences, including two CCoAOMT (EF214740, EF214741) genes, three 4CL (EF214737, EF214738, EF214739) genes, and three F5H (EF214745, EF214746, EF214747) genes. This implies that these are multi-gene families for the genes encoding the three key enzymes in flax lignin biosynthesis. These partial cDNA sequence data are useful for cloning full length cDNA and characterizing the functions and regulations of the critical genes involved in flax lignin biosynthesis.

Key words: Flax (Linum usitatissimum L.), CCoAOMT, 4CL, F5H, Sequence analysis

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