Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China.
College of Life Sciences, University of the Chinese Academy of Sciences, No. 19A Yuquan Road, Shijingshan District, Beijing 100049, China.
Int J Mol Sci. 2021 May 14;22(10):5221. doi: 10.3390/ijms22105221.
The APETALA2 (AP2) transcription factors (TFs) play crucial roles in regulating development in plants. However, a comprehensive analysis of the AP2 family members in a valuable Chinese herbal orchid, , or in other orchids, is limited. In this study, the 14 DoAP2 TFs that were identified from the genome and named DoAP2-1 to DoAP2-14 were divided into three clades: euAP2, euANT, and basalANT. The promoters of all genes contained -regulatory elements related to plant development and also responsive to plant hormones and stress. qRT-PCR analysis showed the abundant expression of , , , and genes in protocorm-like bodies (PLBs), while , , , , and expression was strong in plantlets. In addition, the expression of some genes was down-regulated during flower development. These results suggest that genes may play roles in plant regeneration and flower development in . Four genes ( from euAP2, - from euANT, and and from basal ANT) were selected for further analyses. The transcriptional activation of DoAP2-1, DoAP2-2, DoAP2-6 and DoAP2-11 proteins, which were localized in the nucleus of mesophyll protoplasts, was further analyzed by a dual-luciferase reporter gene system in leaves. Our data showed that pBD-DoAP2-1, pBD-DoAP2-2, pBD-DoAP2-6 and pBD-DoAP2-11 significantly repressed the expression of the LUC reporter compared with the negative control (pBD), suggesting that these DoAP2 proteins may act as transcriptional repressors in the nucleus of plant cells. Our findings on genes in shed light on the function of genes in this orchid and other plant species.
AP2 转录因子（TFs）在植物发育调控中起着至关重要的作用。然而，对一种有价值的中草药或兰花或其他兰花中的 AP2 家族成员进行全面分析的研究是有限的。在本研究中，从 基因组中鉴定出的 14 个 DoAP2 TFs，命名为 DoAP2-1 到 DoAP2-14，分为三个分支：euAP2、euANT 和 basalANT。所有基因的启动子都包含与植物发育相关的 - 调控元件，并且对植物激素和胁迫有反应。qRT-PCR 分析显示，在原球茎样体（PLB）中大量表达 、 、 、 和 基因，而在植物中 、 、 、 、 和 基因的表达较强。此外，一些 基因在花发育过程中表达下调。这些结果表明， 基因可能在 中植物再生和花发育中发挥作用。选择了四个 基因（来自 euAP2 的 、来自 euANT 的 、来自 basal ANT 的 和 ）进行进一步分析。通过双荧光素酶报告基因系统在 叶片中分析了定位于 叶肉原生质体细胞核中的 DoAP2-1、DoAP2-2、DoAP2-6 和 DoAP2-11 蛋白的转录激活。我们的数据表明，与阴性对照（pBD）相比，pBD-DoAP2-1、pBD-DoAP2-2、pBD-DoAP2-6 和 pBD-DoAP2-11 显著抑制了 LUC 报告基因的表达，这表明这些 DoAP2 蛋白可能在植物细胞的细胞核中作为转录抑制剂发挥作用。我们对 基因的研究结果为我们了解该兰花和其他植物物种中 基因的功能提供了线索。
