Characterization of Antirrhinum petal development and identification of target genes of the class B MADS box gene DEFICIENS
Characterization of Antirrhinum petal development and identification of target genes of the class B MADS box gene DEFICIENS
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DOI：
10.1105/tpc.104.026724
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发表时间：
2004-12-01
期刊：
影响因子：
11.6
通讯作者：
Zachgo, S
中科院分区：
文献类型：
--
作者：
Bey, M;Stüber, K;Zachgo, S
金鱼草B类MADS盒转录因子DEF和GLO共同控制花瓣和雄蕊的器官发生。为了了解DEF控制的下游分子机制如何有助于花瓣器官发生，我们使用包含>；11,600个注释金鱼草的大阵列进行了表达谱实验。首先，将花瓣发育后期的四个阶段与花被进行了比较。已鉴定出500多个EST，它们包含大量阶段特异性调控基因，揭示了高度动态的转录调控。为了鉴定可能由DEF直接控制的DEF靶基因，我们利用了对温度敏感的DEF-101突变体。为了提高仿形实验的敏感性，选择了一个花瓣发育阶段，其特征是转录组变化增加，反映了细胞伸长过程而不是细胞分裂过程的开始。在DEF功能降低的情况下，恢复了49个上调和52个下调的花瓣靶基因。通过RT-PCR和原位研究，进一步对8个目的基因进行了详细的鉴定。对DEF活性变化快速反应的基因表达仅限于不同的花瓣组织，这表明DEF功能在花瓣形态发生过程中调节各种基本过程的复杂性。
The class B MADS box transcription factors DEFICIENS (DEF) and GLOBOSA (GLO) of Antirrhinum majus together control the organogenesis of petals and stamens. Toward an understanding of how the downstream molecular mechanisms controlled by DEF contribute to petal organogenesis, we conducted expression profiling experiments using macroarrays comprising >11,600 annotated Antirrhinum unigenes. First, four late petal developmental stages were compared with sepals. More than 500 ESTs were identified that comprise a large number of stage-specifically regulated genes and reveal a highly dynamic transcriptional regulation. For identification of DEF target genes that might be directly controlled by DEF, we took advantage of the temperature-sensitive def-101 mutant. To enhance the sensitivity of the profiling experiments, one petal developmental stage was selected, characterized by increased transcriptome changes that reflect the onset of cell elongation processes replacing cell division processes. Upon reduction of the DEF function, 49 upregulated and 52 downregulated petal target genes were recovered. Eight target genes were further characterized in detail by RT-PCR and in situ studies. Expression of genes responding rapidly toward an altered DEF activity is confined to different petal tissues, demonstrating the complexity of the DEF function regulating diverse basic processes throughout petal morphogenesis.