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作物学报 ›› 2008, Vol. 34 ›› Issue (09): 1563-1568.doi: 10.3724/SP.J.1006.2008.01563

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜油酸脱氢酶基因(fad2)多个拷贝的发现及分析

肖钢1,2;张宏军1,2;彭琪1;官春云1,2,*   

  1. 1 湖南农业大学作物基因工程湖南省重点实验室, 湖南长沙 410128; 2 湖南农业大学油料作物研究所 / 国家油料作物改良中心湖南分中心, 湖南长沙 410128
  • 收稿日期:2007-12-04 修回日期:1900-01-01 出版日期:2008-09-12 网络出版日期:2008-09-12
  • 通讯作者: 官春云

Screening and Analysis of Mutiple Copy of Oleate Desaturase Gene (fad2) in Brassica napus

XIAO Gang12,ZHANG Hong-Jun12,PENG Qi1,GUAN Chun-Yun12*   

  1. 1 Crop Gene Engineering Key Laboratory of Hunan Province, Hunan Agricultural University, Changsha 410128, Hunan; 2 Oil Crops Research Institute, Hunan Agricultural University / Hunan Branch of National Oil Crops Improvement Center, Changsha 410128, Hunan, China
  • Received:2007-12-04 Revised:1900-01-01 Published:2008-09-12 Published online:2008-09-12
  • Contact: GUAN Chun-Yun

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1944

被引次数

35

摘要： 以甘蓝型油菜湘油15为材料, 随机挑选56个来自基因组的fad2基因克隆、47个来自幼苗整株的fad2基因cDNA克隆和9个授粉27 d后种子中fad2 cDNA克隆进行双向测序。基因组中56个fad2序列的碱基同源性为91.0%~99.9%, 从中得到11个差异序列, 即11个不同的fad2基因拷贝。将其翻译成氨基酸序列发现, 6个拷贝在编码区中出现多个终止密码子, 另外5个的同源性为90.60%~99.74%, 与47个来自幼苗整株的fad2基因cDNA序列进行比较, 发现fad2基因没有内含子。从种子中的9个fad2 cDNA克隆序列中找到2个有差异的cDNA, 它们的编码区中没有终止密码子, 说明在种子中有多个fad2基因表达。基因组中的11个拷贝根据同源性可分成两组, 命名为fad2I和fad2II。RT-PCR分析发现在授粉27 d的种子中fad2I有较强表达, fad2II没有表达; 但在叶片中两者都有表达。

关键词: 甘蓝型油菜, 油酸脱氢酶基因, 基因拷贝数, 序列差异引物

Abstract: Rapeseed oil is primarily composed of palmitic, stearic, oleic, linoleic and linolenic fatty acids. Delta-12 desaturase (FAD2) in plants converts oleic acid (18:1) to linoleic acid (18:2) by inserting a double bond at the delta-12 position. Fatty acid desaturase-2 gene (fad2) encodes delta-12 desaturase that functions in the endoplasmic reticulum. Fifty-six fad2 DNA clones and nine fad2 seed cDNA clones of B. napus cv. Xiangyou 15 were randomly selected and sequenced in this study. These 56 DNA sequences share 91.0-99.9% identity in nucleotides. Through combination of identical sequences, 11 different sequences were found, of which each represented various copy of fad2 gene. Deduced amino acid sequences revealed that many stop codons occurred in the coding region of six copies, the other five copies shared 90.60-99.74% amino-acid identity. Two differential cDNA were found from nine fad2 seed cDNA sequence, no stop codons in the coding region, revealing that more than one fad2 gene express in developing seeds, and oleic acid content is controlled by multigene. The eleven copies in genome could be divided into two groups based on their homology, and designated as fad2I and fad2II, respectively. RT-PCR analysis in seeds at twenty-seven days after pollination showed that fad2I expressed strongly in seeds, and no expression of fad2II was found, but both of them expressed in leaves.

Key words: Brassica napus, Oleate desaturase, Gene copy number, Sequence-specific primer

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