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为了研究WRKY基因在香蕉与Foc TR4(Fusarium oxysporum f.sp.cubense race 4)互作中作用,利用RACE技术从香蕉中克隆获得了一个WRKY基因全长。序列分析表明,该基因包含完整的开放阅读框,编码275个氨基酸,与香蕉A基因组预测出的(XP_009392153)成员的同源性为100%,因此将其命名为MaWRKY18。MaWRKY18与MaWRKY71(ADR71866)亲缘关系近,聚类在一个分支上。RT-PCR分析表明该基因在香蕉各器官(根、球茎、假茎、叶、花和果实)中均有表达。MaWRKY18受水杨酸和乙烯诱导上调表达,受茉莉酸诱导下调表达。亚细胞定位表明该基因编码的蛋白定位在细胞核中,且C端具有转录激活活性。在Foc TR4接种后,MaWRKY18在感病品种中下调表达,在抗病品种中上调表达。结果表明克隆到的MaWRKY18可能在香蕉与Foc TR4互作过程中介导抗病性。
In order to study the role of WRKY in the interaction of banana with Foc TR4 (Fusarium oxysporum f. Sp.cubense race 4), a full-length WRKY gene was cloned from banana using RACE technique. Sequence analysis showed that this gene contains a complete open reading frame and encodes a protein of 275 amino acids, which is named as MaWRKY18 because it has 100% homology with the predicted members of the banana A genome (XP_009392153). MaWRKY18 has a close genetic relationship with MaWRKY71 (ADR71866), clustering on one branch. RT-PCR analysis showed that the gene was expressed in all organs of banana (root, bulb, pseudostem, leaf, flower and fruit). MaWRKY18 was up-regulated by salicylic acid and ethylene and down-regulated by jasmonic acid. Subcellular localization indicated that the protein encoded by this gene was located in the nucleus and the C-terminal had transcriptional activation activity. After Foc TR4 inoculation, MaWRKY18 was down-regulated in susceptible varieties and up-regulated in resistant varieties. The results showed that the cloned MaWRKY18 may mediate disease resistance during banana interaction with Foc TR4.