De Paolis Angelo, Caretto Sofia, Quarta Angela, Di Sansebastiano Gian-Pietro, Sbrocca Irene, Mita Giovanni, Frugis Giovanna
Istituto di Scienze delle Produzioni Alimentari (ISPA), Consiglio Nazionale delle Ricerche (CNR), Via Monteroni, 73100 Lecce, Italy.
DiSTeBA (Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali), University of Salento, Campus ECOTEKNE, 73100 Lecce, Italy.
Plants (Basel). 2020 Nov 28;9(12):1669. doi: 10.3390/plants9121669.
L. is well-known as the plant source of artemisinin, a sesquiterpene lactone with effective antimalarial activity. Here, a putative ortholog of the WRKY40 transcription factor (TF) was isolated via reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends in and named . A putative nuclear localization domain was identified in silico and experimentally confirmed by using protoplasts of transiently transformed with . A genome-wide analysis identified 122 genes in , and a manually curated database was obtained. The deduced proteins were categorized into the major WRKY groups, with group IIa containing eight WRKY members including AaWRKY40. Protein motifs, gene structure, and promoter regions of group IIa WRKY TFs of were characterized. The promoter region of group IIa genes contained several abiotic stress -acting regulatory elements, among which a highly conserved W-box motif was identified. Expression analysis of compared to in cell cultures treated with methyl jasmonate known to enhance artemisinin production, suggested a possible involvement of in terpenoid metabolism. Further investigation is necessary to study the role of AaWRKY40 and possible interactions with other TFs in .
黄花蒿是青蒿素的植物来源，青蒿素是一种具有有效抗疟活性的倍半萜内酯。在此，通过逆转录聚合酶链反应和cDNA末端快速扩增，从黄花蒿中分离出WRKY40转录因子（TF）的一个假定直系同源物，并将其命名为AaWRKY40。通过计算机分析鉴定出一个假定的核定位结构域，并通过用AaWRKY40瞬时转化的原生质体进行实验验证。全基因组分析在黄花蒿中鉴定出122个WRKY基因，并获得了一个人工整理的数据库。推导的蛋白质被归类到主要的WRKY组中，IIa组包含八个WRKY成员，包括AaWRKY40。对黄花蒿IIa组WRKY转录因子的蛋白质基序、基因结构和启动子区域进行了表征。黄花蒿IIa组基因的启动子区域包含几个非生物胁迫作用的调控元件，其中鉴定出一个高度保守的W-box基序。在用已知可提高青蒿素产量的茉莉酸甲酯处理的黄花蒿细胞培养物中，与对照相比对AaWRKY40的表达分析表明，AaWRKY40可能参与萜类代谢。有必要进一步研究AaWRKY40的作用以及它与黄花蒿中其他转录因子可能的相互作用。
