J Irepan Reyes-Olalde1, Nayelli Marsch-Martínez2, Stefan de Folter1. 1. Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Irapuato, Guanajuato, México. 2. Departamento de Biotecnología y Bioquímica, CINVESTAV-IPN, Irapuato, Guanajuato, México.
Abstract
BACKGROUND: The gynoecium is the female reproductive structure and probably the most complex plant structure. During its development, different internal tissues and structures are formed. Insights in gene expression or hormone localization patterns are key to understanding gynoecium development from a molecular biology point of view. RESULTS: Imaging with a confocal laser scanning microscope (CLSM) is a widely used strategy; however, visualization of internal developmental expression patterns in the Arabidopsis gynoecium can be technically challenging. Here, we present a detailed protocol that allows the visualization of internal expression patterns at high resolution during gynoecium development. We demonstrate the applicability using a cytokinin response marker (TCS::GFP), an auxin response marker (DR5::VENUS), and a SEPALLATA3 marker (SEP3::SEP3:GFP). CONCLUSIONS: The detailed protocol presented here allows the visualization of fluorescence signals in internal structures during Arabidopsis gynoecium development. This protocol may also be adapted for imaging other challenging plant structures or organs.
BACKGROUND: The gynoecium is the female reproductive structure and probably the most complex plant structure. During its development, different internal tissues and structures are formed. Insights in gene expression or hormone localization patterns are key to understanding gynoecium development from a molecular biology point of view. RESULTS: Imaging with a confocal laser scanning microscope (CLSM) is a widely used strategy; however, visualization of internal developmental expression patterns in the Arabidopsis gynoecium can be technically challenging. Here, we present a detailed protocol that allows the visualization of internal expression patterns at high resolution during gynoecium development. We demonstrate the applicability using a cytokinin response marker (TCS::GFP), an auxin response marker (DR5::VENUS), and a SEPALLATA3 marker (SEP3::SEP3:GFP). CONCLUSIONS: The detailed protocol presented here allows the visualization of fluorescence signals in internal structures during Arabidopsis gynoecium development. This protocol may also be adapted for imaging other challenging plant structures or organs.
Authors: J Irepan Reyes-Olalde; Víctor M Zúñiga-Mayo; Joanna Serwatowska; Ricardo A Chavez Montes; Paulina Lozano-Sotomayor; Humberto Herrera-Ubaldo; Karla L Gonzalez-Aguilera; Patricia Ballester; Juan José Ripoll; Ignacio Ezquer; Dario Paolo; Alexander Heyl; Lucia Colombo; Martin F Yanofsky; Cristina Ferrandiz; Nayelli Marsch-Martínez; Stefan de Folter Journal: PLoS Genet Date: 2017-04-07 Impact factor: 5.917
Authors: Hugo G Lazcano-Ramírez; Andrea Gómez-Felipe; David Díaz-Ramírez; Yolanda Durán-Medina; Lino Sánchez-Segura; Stefan de Folter; Nayelli Marsch-Martínez Journal: Front Plant Sci Date: 2018-09-25 Impact factor: 5.753