玉米ZmFAR1基因的克隆与表达分析

Cloning and Expression Analysis of ZmFAR1 Gene in Maize

FARs为脂酰辅酶A还原酶,在植物生长发育和抵御非生物胁迫过程中起着重要作用。为进一步研究FARs在玉米中行使的生物学功能,在玉米中克隆1个脂酰辅酶A还原酶1基因(ZmFAR1),该基因全长1883 bp,开放阅读框长1491 bp,编码496个氨基酸,分子量为55.983 k Da。生物信息学分析表明,ZmFAR1蛋白的分子式为C_(2522)H_(4010)N^(690)O_(701)S_(24),为稳定的碱性亲水蛋白,存在59个磷酸化位点,未发现信号肽,存在跨膜结构域,最大可能定位于细胞质。氨基酸残基组成中主要以α螺旋和无规则卷曲为主。系统进化树分析表明,ZmFAR1蛋白与南荻和高粱的FAR蛋白相似度最高。利用实时荧光定量PCR技术对ZmFAR1在模拟盐、干旱两种非生物胁迫条件下对根、茎、叶3种组织的表达模式进行研究,结果表明,ZmFAR1呈现组织特异性表达,在叶中的表达量最高。盐和干旱处理下,ZmFAR1的表达量出现明显变化,推测ZmFAR1可能在不同程度参与了玉米对非生物胁迫的应答。

Fatty acyl-CoA reductases(FARs)play an important role in the process of plant growth and develop-ment and resistance to abiotic stress.The cDNA sequence of ZmFARI gene from maize was cloned with a full length of 1883 bp and an open reading frame of 1491 bp,encoding 496 amino acids with molecular weights of 55.983 kDa.The results of bioinformatics analysis showed that the molecular formula of ZmFAR1 protein is C_(2522)H_(4010)N_(690)O_(701)S_(24),which is a stable basic hydrophilic protein with 59 phosphorylation sites and no signal peptide.And there is a transmembrane domain,which is the most likely to be located in the cytoplasm.The composition of amino acid residues is mainly composed of the alpha helix and random coil.The evolutionary relationship showed that the ZmFAR1 protein was related with FAR from Miscanthus(Miscanthus Lutarioriparius)and sorghum(Sorghum bicolor).At the same time,qRT-PCR was used to study its expression patterns in three tissues of root,stem and leaf and under two abiotic stress conditions of salt and drought.The results showed that the expression of ZmFARI was tissue-specific,and there is the highest expression in leaves.After salt and simulated drought treatments,the expression of ZmFARI changed significantly.It is speculated that ZmFARI may participate in the response of maize to abiotic stress in different degrees.