Krizek Beth A
Department of Biological Sciences, University of South Carolina, Columbia, SC, 29208, USA.
BMC Res Notes. 2015 Oct 12;8:556. doi: 10.1186/s13104-015-1537-6.
The AINTEGUMENTA-LIKE6/PLETHORA3 (AIL6/PLT3) gene of Arabidopsis thaliana is a key regulator of growth and patterning in both shoots and roots. AIL6 encodes an AINTEGUMENTA-LIKE/PLETHORA (AIL/PLT) transcription factor that is expressed in the root stem cell niche, the peripheral region of the shoot apical meristem and young lateral organ primordia. In flowers, AIL6 acts redundantly with AINTEGUMENTA (ANT) to regulate floral organ positioning, growth, identity and patterning. Experiments were undertaken to define the genomic regions required for AIL6 function and expression in flowers.
Transgenic plants expressing a copy of the coding region of AIL6 in the context of 7.7 kb of 5' sequence and 919 bp of 3' sequence (AIL6:cAIL6-3') fail to fully complement AIL6 function when assayed in the ant-4 ail6-2 double mutant background. In contrast, a genomic copy of AIL6 with the same amount of 5' and 3' sequence (AIL6:gAIL6-3') can fully complement ant-4 ail6-2. In addition, a genomic copy of AIL6 with 590 bp of 5' sequence and 919 bp of 3' sequence (AIL6m:gAIL6-3') complements ant-4 ail6-2 and contains all regulatory elements needed to confer normal AIL6 expression in inflorescences. Efforts to map cis-regulatory elements reveal that the third intron of AIL6 contains enhancer elements that confer expression in young flowers but in a broader pattern than that of AIL6 mRNA in wild-type flowers. Some AIL6:gAIL6-3' and AIL6m:gAIL6-3' lines confer an over-rescue phenotype in the ant-4 ail6-2 background that is correlated with higher levels of AIL6 mRNA accumulation.
The results presented here indicate that AIL6 intronic sequences serve as transcriptional enhancer elements. In addition, the results show that increased expression of AIL6 can partially compensate for loss of ANT function in flowers.
拟南芥的类珠被6/多胚座3(AIL6/PLT3)基因是地上部分和根部生长及模式形成的关键调节因子。AIL6编码一种类珠被/多胚座(AIL/PLT)转录因子,该因子在根干细胞龛、茎尖分生组织的周边区域以及幼嫩侧生器官原基中表达。在花中,AIL6与珠被(ANT)发挥冗余作用,以调节花器官的定位、生长、特性及模式形成。开展实验以确定AIL6在花中的功能及表达所需的基因组区域。
在7.7 kb的5'序列和919 bp的3'序列背景下表达AIL6编码区拷贝的转基因植物(AIL6:cAIL6-3'),在ant-4 ail6-2双突变体背景下进行检测时,未能完全恢复AIL6的功能。相比之下,具有相同数量5'和3'序列的AIL6基因组拷贝(AIL6:gAIL6-3')能够完全恢复ant-4 ail6-2的功能。此外,具有590 bp的5'序列和919 bp的3'序列的AIL6基因组拷贝(AIL6m:gAIL6-3')能恢复ant-4 ail6-2的功能,且包含在花序中赋予正常AIL6表达所需的所有调控元件。对顺式调控元件进行定位的研究表明,AIL6的第三个内含子包含增强子元件,这些元件能在幼嫩花中赋予表达,但表达模式比野生型花中AIL6 mRNA的表达模式更广泛。一些AIL6:gAIL6-3'和AIL6m:gAIL6-3'株系在ant-4 ail6-2背景下表现出过度拯救表型,这与更高水平的AIL6 mRNA积累相关。
此处呈现的结果表明,AIL6内含子序列充当转录增强子元件。此外,结果显示AIL6表达的增加可部分补偿花中ANT功能的丧失。