RESUMEN
Plants flower in response to environmental signals. These signals change the shape and developmental identity of the shoot apical meristem (SAM), causing it to form flowers and inflorescences. We show that the increases in SAM width and height during floral transition correlate with changes in size of the central zone (CZ), defined by CLAVATA3 expression, and involve a transient increase in the height of the organizing center (OC), defined by WUSCHEL expression. The APETALA2 (AP2) transcription factor is required for the rapid increases in SAM height and width, by maintaining the width of the OC and increasing the height and width of the CZ. AP2 expression is repressed in the SAM at the end of floral transition, and extending the duration of its expression increases SAM width. Transcriptional repression by SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) represents one of the mechanisms reducing AP2 expression during floral transition. Moreover, AP2 represses SOC1 transcription, and we find that reciprocal repression of SOC1 and AP2 contributes to synchronizing precise changes in meristem shape with floral transition.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Flores , Regulación de la Expresión Génica de las Plantas , Proteínas de Homeodominio , Proteínas de Dominio MADS , Meristema , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Meristema/metabolismo , Meristema/crecimiento & desarrollo , Meristema/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Flores/genética , Flores/crecimiento & desarrollo , Flores/metabolismo , Proteínas de Homeodominio/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Dominio MADS/metabolismo , Proteínas de Dominio MADS/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Regulación del Desarrollo de la Expresión Génica , Plantas Modificadas GenéticamenteRESUMEN
The APETALA2 (AP2) transcription factor regulates flower development, floral transition and shoot apical meristem (SAM) maintenance in Arabidopsis. AP2 is also regulated at the post-transcriptional level by microRNA172 (miR172), but the contribution of this to SAM maintenance is poorly understood. We generated transgenic plants carrying a form of AP2 that is resistant to miR172 (rAP2) or carrying a wild-type AP2 susceptible to miR172. Phenotypic and genetic analyses were performed on these lines and mir172 mutants to study the role of AP2 regulation by miR172 on meristem size and the rate of flower production. We found that rAP2 enlarges the inflorescence meristem by increasing cell size and cell number. Misexpression of rAP2 from heterologous promoters showed that AP2 acts in the central zone (CZ) and organizing center (OC) to increase SAM size. Furthermore, we found that AP2 is negatively regulated by AUXIN RESPONSE FACTOR 3 (ARF3). However, genetic analyses indicated that ARF3 also influences SAM size and flower production rate independently of AP2. The study identifies miR172/AP2 as a regulatory module controlling inflorescence meristem size and suggests that transcriptional regulation of AP2 by ARF3 fine-tunes SAM size determination.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , MicroARNs , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Flores , Regulación de la Expresión Génica de las Plantas , Proteínas de Homeodominio/metabolismo , Inflorescencia/metabolismo , Meristema/metabolismo , MicroARNs/genética , Proteínas Nucleares/metabolismoRESUMEN
Plant roots adapt their development and metabolism to changing environmental conditions. In order to understand the response mechanisms of roots to the dynamic availability of water or nutrients, to biotic and abiotic stress conditions or to mechanical stimuli, microfluidic platforms have been developed that offer microscopic access and novel experimental means. Here, we describe the design, fabrication and use of microfluidic devices suitable for imaging growing Arabidopsis roots over several days under controlled perfusion. We present a detailed protocol for the use of our exemplar platform-the RootChip-8S-and offer a guide for troubleshooting, which is also largely applicable to related device designs. We further discuss considerations regarding the design of custom-made plant microdevices, the choice of suitable materials and technologies as well as the handling of the specimen.