title: An efficient method for miRNA detection and localization in crop plants.
abstract: microRNAs are a class of non-coding small RNAs (sRNAs) that are important regulators of gene expression at the post-transcriptional level by mRNA cleavage or translation inhibition. Another class of sRNAs are siRNAs, which also regulate gene expression but by causing DNA methylation. This epigenetic regulatory role has been observed for some miRNAs as well. The use of sRNAs allows the development of biotechnological applications in plants. To develop these types of applications, and to better understand the natural roles they play, it is important to be able to detect and to localize these sRNAs at the plant tissue level. Sometimes, in crop plants this can be challenging. Therefore, we developed a tissue printing hybridization protocol for easy and efficient detection of sRNAs and demonstrate this by the analysis of the spatio-temporal expression patterns of the miRNAs miR159 and miR164 in fruits of various crop plants. Moreover, we show the possibility to also detect the expression of miRNAs in fruit juice using a dot blot hybridization approach.
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Guanajuato, México ; Centro de Investigación en Biotecnología Aplicada del Instituto Politécnico Nacional Tlaxcala, México.
authors.forename: Flor de Fátima
authors.initials: Fde F
authors.lastname: Rosas-Cárdenas
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Guanajuato, México.
authors.forename: Rocío
authors.initials: R
authors.lastname: Escobar-Guzmán
authors.affiliation: Facultad de Ingeniería, Universidad Autónoma de Querétaro Querétaro, México.
authors.forename: Andrés
authors.initials: A
authors.lastname: Cruz-Hernández
authors.affiliation: Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Guanajuato, México.
authors.forename: Nayelli
authors.initials: N
authors.lastname: Marsch-Martínez
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional Guanajuato, México.
authors.forename: Stefan
authors.initials: S
authors.lastname: de Folter
keywords: fruits; hybridization; miR164; miRNA detection; tissue-printing