油葵HaFAD22基因的克隆及表达分析
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国家自然科学基金(31360052);


Cloning and Expression of HaFAD22 Gene from Helianthus annuus L.
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    摘要:

    脂肪酸脱氢酶2(fatty acid desaturase, FAD2)催化油酸生成亚油酸,是植物体内生成多不饱和脂肪酸的关键酶。根据已报道的向日葵(Helianthus annuus L.)FAD2基因序列,设计引物进行RTPCR,克隆得到油葵FAD22基因全长cDNA,命名为HaFAD22。该基因开放阅读框为1 152 bp,编码383个氨基酸,相对分子质量43.96 kD,等电点为8.56。对基因组进行内含子调查发现,该基因在编码区内没有内含子。多序列比对和系统进化分析发现,FAD22基因编码蛋白与金盏菊(Calendula officinalis)、斑鸠菊(Vernonia galamensis)等菊科植物具有较近的亲缘关系。qRTPCR分析表明,HaFAD22基因在根、茎、叶、花、子叶和未成熟种子中均有表达,且以叶中的表达量最高,未成熟种子中的表达量最低;低温(5 ℃、15 ℃)胁迫处理能显著促进该基因在根中的表达,抑制其在叶中的表达;盐胁迫(300 mmol/L NaCl)处理对其表达也具有抑制作用。该研究结果可为进一步探讨HaFAD22基因的功能奠定基础。

    Abstract:

    Linoleic acid is synthesized by fatty acid desaturase 2(FAD2), which play an important role for produce polyunsaturated fatty acid in plant. The key enzyme for the conversion of oleic acid into linoleic acid was isolated from sunflower (Helianthus annuus L.), and designated as HaFAD22.The open reading frame (ORF) of HaFAD22 was 1 152 bp in length, which encoded 383 amino acids, with the isoelectric point of 8.56 and molecular mass of 43.96 kD. The genome intron survey showed that there is no introns in open reading frame region. Phylogenetic analysis showed that HaFAD22 protein shares high homologies with calendula (Calendula officinalis) and dove (Vernonia galamensis). qRTPCR analysis indicated that HaFAD22 was expressed in roots, stems, leaves, cotyledon, flowers and developing seeds, and the highest in leaves, the lowest in developing seeds. Furthermore, HaFAD22 in roots could be induced significantly by low temperature (5 ℃, 15 ℃), but repressed in leaves by low temperature (5 ℃, 15 ℃) and NaCl stress (300 mmol/L). Results in this experiment provide reference for further functional study on HaFAD22.

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周茜萍,王梦瑶,冯 丽,等.油葵HaFAD22基因的克隆及表达分析[J].西北植物学报,2017,37(5):845-850

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  • 在线发布日期: 2017-06-05
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