红掌交替氧化酶基因克隆及其在低温胁迫下的表达分析

Molecular Cloning and Low Temperature Induced Expression of an Alternative Oxidase Gene from Anthurium andraeanum

以天南星科花烛属红掌阿拉巴马叶片提取的总RNA为模板,通过RT-PCR与RACE扩增,获得一个1170 bp的交替氧化酶(Alternative oxidase,AOX)基因的cDNA序列,其基因编码区共1035 bp,编码345个氨基酸,命名为AnAOX(GenBank登录号:JX163297)。用低温胁迫处理红掌植株,叶片最大光化学效率Fv/Fm随着低温胁迫处理时间的延长而逐渐下降,当6℃处理达到36 h时Fv/Fm开始急剧下降,叶片也出现萎蔫状态,表明红掌叶片受到极大的伤害。使用实时荧光定量PCR,对不同低温处理时间下AOX基因在红掌植物叶片中的表达量进行测定分析,结果表明:低温胁迫处理后,AnAOX基因均上调表达,且处理24 h后表达量达到最高,表明其与红掌抗低温胁迫有一定的关系。

Using the total RNA from Anthurium andraeanum ＂Alabama＂leaves as the template, a full-length cDNA of alternative oxidase （AOX） gene was obtained by RT-PCR and RACE. Bioinformatics analysis indicated that the full- length of cDNA sequence was 1170 bp, which contained an open reading frame of 1035 bp and encoded a protein of 345 amino acid residues. The maximum photochemical efficiency of PSII （Fv/Fm） was decreased along with the time course of cold stress, withering appeared on the leaves, and the Fv/Fm was also decreased sharply when the plants were treated at 6 ~C for 36 h~ This suggested that the plant was greatly injured. Real-time PCR analysis showed that the expression of AnAOX was up-regulated under different time course of cold stress, and the relative mRNA amount reached the peak after treated at low temperature for 24 h. These results suggested that AnAOX was involved in cold resistant in young leaves of Anthurium andraeanum.