Meiocyte Isolation by INTACT and Meiotic Transcriptome Analysis in Arabidopsis
| Autor: | Gaetana Cremona, Clara Conicella, Maria Federica Consiglio, Carmine Lanzillo, Lucia Barra, Riccardo Aiese Cigliano, Rosa Paparo, Pasquale Termolino |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Frontiers in Plant Science Frontiers in plant science 12 (2021). doi:10.3389/fpls.2021.638051 info:cnr-pdr/source/autori:Barra Lucia 1, Termolino Pasquale 1, Aiese Cigliano Riccardo 2, Cremona Gaetana 1, Paparo Rosa 1, Lanzillo Carmine 1, Consiglio Maria Federica 1, Conicella Clara 1/titolo:Meiocyte Isolation by INTACT and Meiotic Transcriptome Analysis in Arabidopsis/doi:10.3389%2Ffpls.2021.638051/rivista:Frontiers in plant science/anno:2021/pagina_da:/pagina_a:/intervallo_pagine:/volume:12 Frontiers in Plant Science, Vol 12 (2021) |
| ISSN: | 1664-462X |
| DOI: | 10.3389/fpls.2021.638051 |
| Popis: | Isolation of nuclei tagged in specific cell types (INTACT) is a method developed to isolate cell-type-specific nuclei that are tagged through in vivo biotin labeling of a nuclear targeting fusion (NTF) protein. In our work, INTACT was used to capture nuclei of meiocytes and to generate a meiotic transcriptome in Arabidopsis. Using the promoter of AtDMC1 recombinase to label meiotic nuclei, we generated transgenic plants carrying AtDMC1:NTF along with biotin ligase enzyme (BirA) under the constitutive ACTIN2 (ACT2) promoter. AtDMC1-driven expression of biotin-labeled NTF allowed us to collect nuclei of meiocytes by streptavidin-coated magnetic beads. The nuclear meiotic transcriptome was obtained by RNA-seq using low-quantity input RNA. Transcripts grouped into different categories according to their expression levels were investigated by gene ontology enrichment analysis (GOEA). The most enriched GO term “DNA demethylation” in mid/high-expression classes suggests that this biological process is particularly relevant to meiosis onset. The majority of genes with established roles in meiosis were distributed in the classes of mid/high and high expression. Meiotic transcriptome was compared with public available transcriptomes from other tissues in Arabidopsis. Bioinformatics analysis by expression network identified a core of more than 1,500 genes related to meiosis landmarks. |
| Databáze: | OpenAIRE |
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