de Moura Stéfanie Menezes, Artico Sinara, Lima Cássio, Nardeli Sarah Muniz, Berbel Ana, Oliveira-Neto Osmundo Brilhante, Grossi-de-Sá Maria Fátima, Ferrándiz Cristina, Madueño Francisco, Alves-Ferreira Márcio
Department of Genetics, Universidade Federal do Rio de Janeiro (UFRJ), Av. Prof. Rodolpho Paulo Rocco, s/n - Prédio do CCS - Instituto de Biologia, 2° andar, sala A2-93, Rio de Janeiro, RJ, 219410-970, Brazil.
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas - Universidad Politécnica de Valencia, Valencia, Spain.
Plant Reprod. 2017 Mar;30(1):19-39. doi: 10.1007/s00497-017-0297-y. Epub 2017 Feb 7.
Expression analysis of the AG -subfamily members from G. hirsutum during flower and fruit development. Reproductive development in cotton, including the fruit and fiber formation, is a complex process; it involves the coordinated action of gene expression regulators, and it is highly influenced by plant hormones. Several studies have reported the identification and expression of the transcription factor family MADS-box members in cotton ovules and fibers; however, their roles are still elusive during the reproductive development in cotton. In this study, we evaluated the expression profiles of five MADS-box genes (GhMADS3, GhMADS4, GhMADS5, GhMADS6 and GhMADS7) belonging to the AGAMOUS-subfamily in Gossypium hirsutum. Phylogenetic and protein sequence analyses were performed using diploid (G. arboreum, G. raimondii) and tetraploid (G. barbadense, G. hirsutum) cotton genomes, as well as the AG-subfamily members from Arabidopsis thaliana, Petunia hybrida and Antirrhinum majus. qPCR analysis showed that the AG-subfamily genes had high expression during flower and fruit development in G. hirsutum. In situ hybridization analysis also substantiates the involvement of AG-subfamily members on reproductive tissues of G. hirsutum, including ovule and ovary. The effect of plant hormones on AG-subfamily genes expression was verified in cotton fruits treated with gibberellin, auxin and brassinosteroid. All the genes were significantly regulated in response to auxin, whereas only GhMADS3, GhMADS4 and GhMADS7 genes were also regulated by brassinosteroid treatment. In addition, we have investigated the GhMADS3 and GhMADS4 overexpression effects in Arabidopsis plants. Interestingly, the transgenic plants from both cotton AG-like genes in Arabidopsis significantly altered the fruit size compared to the control plants. This alteration suggests that cotton AG-like genes might act regulating fruit formation. Our results demonstrate that members of the AG-subfamily in G. hirsutum present a conserved expression profile during flower development, but also demonstrate their expression during fruit development and in response to phytohormones.
陆地棉AG亚家族成员在花和果实发育过程中的表达分析。棉花的生殖发育，包括果实和纤维形成，是一个复杂的过程；它涉及基因表达调节因子的协同作用，并且受到植物激素的高度影响。几项研究报道了棉花胚珠和纤维中转录因子家族MADS-box成员的鉴定和表达；然而，它们在棉花生殖发育过程中的作用仍然不清楚。在本研究中，我们评估了陆地棉中属于AGAMOUS亚家族的五个MADS-box基因（GhMADS3、GhMADS4、GhMADS5、GhMADS6和GhMADS7）的表达谱。使用二倍体（亚洲棉、雷蒙德氏棉）和四倍体（海岛棉、陆地棉）棉花基因组，以及拟南芥、矮牵牛和金鱼草的AG亚家族成员进行了系统发育和蛋白质序列分析。qPCR分析表明，AG亚家族基因在陆地棉的花和果实发育过程中具有高表达。原位杂交分析也证实了AG亚家族成员参与陆地棉的生殖组织，包括胚珠和子房。在用赤霉素、生长素和油菜素内酯处理的棉花果实中验证了植物激素对AG亚家族基因表达的影响。所有基因对生长素处理均有显著调控，而只有GhMADS3、GhMADS4和GhMADS7基因也受油菜素内酯处理的调控。此外，我们研究了GhMADS3和GhMADS4在拟南芥植物中的过表达效应。有趣的是，与对照植物相比，拟南芥中两个棉花AG类基因的转基因植物显著改变了果实大小。这种改变表明棉花AG类基因可能在调节果实形成中起作用。我们的结果表明，陆地棉AG亚家族成员在花发育过程中呈现保守的表达谱，但也证明了它们在果实发育过程中的表达以及对植物激素的响应。
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