Department of Biology, University of Leicester, University Road, Leicester LE1 7RH, UK.
Ann Bot. 2011 Jun;107(9):1557-66. doi: 10.1093/aob/mcr045. Epub 2011 Mar 7.
The MADS-box transcription factor AGAMOUS (AG) is an important regulator of stamen and fruit identity as well as floral meristem determinacy in a number of core eudicots and monocots. However, its role outside of these groups has not been assessed explicitly. Examining its role in opium poppy, a basal eudicot, could uncover much about the evolution and development of flower and fruit development in the angiosperms.
AG orthologues were isolated by degenerate RT-PCR and the gene sequence and structure examined; gene expression was characterized using in situ hybridization and the function assessed using virus-induced gene silencing.
In opium poppy, a basal eudicot, the AGAMOUS orthologue is alternatively spliced to produce encoded products that vary at the C-terminus, termed PapsAG-1 and PapsAG-2. Both transcripts are expressed at high levels in stamens and carpels. The functional implications of this alternative transcription were examined using virus-induced gene silencing and the results show that PapsAG-1 has roles in stamen and carpel identity, reflecting those found for Arabidopsis AG. In contrast, PapsAG-2, while displaying redundancy in these functions, has a distinctive role in aspects of carpel development reflected in septae, ovule and stigma defects seen in the loss-of-function line generated.
These results describe the first explicit functional analysis of an AG-clade gene in a basal eudicot; illustrate one of the few examples of the functional consequences of alternative splicing in transcription factors and reveal the importance of alternative transcription, as well as gene duplication, as a driving force in evolution.
MADS-box 转录因子 AGAMOUS(AG)是多个核心真双子叶植物和单子叶植物雄蕊和果实身份以及花分生组织确定性的重要调节因子。然而,其在这些植物群之外的作用尚未被明确评估。在罂粟中研究其作用,一种基础真双子叶植物,可以揭示花和果实发育在被子植物中的进化和发育。
通过简并 RT-PCR 分离 AG 同源物,并检查基因序列和结构;使用原位杂交进行基因表达特征分析,并使用病毒诱导的基因沉默评估其功能。
在罂粟中,一种基础真双子叶植物,AGAMOUS 同源物通过选择性剪接产生在 C 末端变化的编码产物,称为 PapsAG-1 和 PapsAG-2。这两种转录本在雄蕊和心皮中都高表达。使用病毒诱导的基因沉默来检查这种选择性转录的功能意义,结果表明 PapsAG-1 在雄蕊和心皮身份中具有作用,反映了拟南芥 AG 中的作用。相比之下,PapsAG-2 在这些功能中具有冗余性,但在心皮发育的某些方面具有独特的作用,表现在功能丧失系中看到的隔膜、胚珠和柱头缺陷中。
这些结果描述了在基础真双子叶植物中对 AG 类基因进行的首次明确的功能分析;说明了转录因子选择性剪接在功能上的少数几个例子之一,并揭示了替代转录以及基因复制作为进化驱动力的重要性。