摘要
采用同源克隆法从菜心中获得cDNA全长459 bp的E2基因,命名为BclUBE2。该基因编码153个氨基酸,分子量为17.24ku,理论等电点为5.37,为亲水性非分泌蛋白。结构分析显示,该蛋白含有一个泛素结合酶E2活性位点、一个WD重复序列和一个高度保守的半胱氨酸。进化分析发现,菜心BclUBE2蛋白与芸薹属芜菁和油菜的亲缘关系较近。qRT-PCR分析表明,菜心BclUBE2基因在根、茎、叶、叶柄中均有表达,且表达丰度和变化趋势不同。在低温胁迫下,根中的表达量呈现先降低后升高的变化趋势,且在处理1 h时表达量最低;在茎、叶、叶柄中均呈现先升高后降低的表达趋势,茎和叶柄处理6 h时的表达量最高,叶片处理1 h时的表达量最高。说明菜心BclUBE2基因在响应低温胁迫中发挥作用。
The ubiquitin-conjugating enzyme E2 gene was cloned from Brassica campestris L. ssp. chinensis var. utilisTsen et Lee and designated as BclUBE2. The full cDNA length of BclUBE2 was 459 bp, encoding 153 amino acid residues with an estimated molecular mass of 17.24 ku and a calculated pI of 5.37. This protein was hydrophilic non-secretory. Structural analysis revealed that the sequence contained an E2 ubiqitin-conjugating enzyme active site, a WD-repeated sequence and a highly conserved cysteine. Phylogenetic analysis showed that BclUBE2 protein had the closest relationship with Brassica napus and Brassica rapa. qRT-PCR results showed that the existence of BclUBE2 was detectable in the root, stem, leaf and petiole with different expressive abundance and change trend. Under low temperature stress, the expression level in the root showed the trend of decreasing first and then increasing, and reached the lowest level at 1 h. However, the expression level in the stem, leaf and petiole increased first and then decreased with the highest level at 6 h in stem and petiole and at 1 h in leaf. These results indicated that BclUBE2 gene was involved in plant response to low temperature stress.
作者
曾小玲
赵瑞丽
钟开勤
朱朝辉
陈敏氡
ZENG Xiaoling;ZHAO Ruili;ZHONG Kaiqin;ZHU Chaohui;CHEN Mindong(Fuzhou Institute of Vegetable Science,Fuzhou,Fujian 350111,China;Crops Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350013,China)
出处
《热带作物学报》
CSCD
北大核心
2018年第9期1772-1777,共6页
Chinese Journal of Tropical Crops
基金
福州市重大专项(No.2016Z0001)
福建省星火项目(No.2016S0006)
关键词
菜心
BclUBE2基因
QRT-PCR
Brassica campestris L. ssp.chinensis var. utilis Tsen et Lee
BclUBE2 gene
qRT-PCR
