AUTHOR=Barra Lucia , Termolino Pasquale , Aiese Cigliano Riccardo , Cremona Gaetana , Paparo Rosa , Lanzillo Carmine , Consiglio Maria Federica , Conicella Clara 

TITLE=Meiocyte Isolation by INTACT and Meiotic Transcriptome Analysis in Arabidopsis

JOURNAL=Frontiers in Plant Science

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.638051

DOI=10.3389/fpls.2021.638051

ISSN=1664-462X

ABSTRACT=<p>Isolation of nuclei tagged in specific cell types (INTACT) is a method developed to isolate cell-type-specific nuclei that are tagged through <italic>in vivo</italic> 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 <italic>Arabidopsis</italic>. Using the promoter of <italic>AtDMC1</italic> recombinase to label meiotic nuclei, we generated transgenic plants carrying <italic>AtDMC1:NTF</italic> along with biotin ligase enzyme (<italic>BirA</italic>) under the constitutive <italic>ACTIN2</italic> (<italic>ACT2</italic>) promoter. <italic>AtDMC1</italic>-driven expression of biotin-labeled <italic>NTF</italic> 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.</p>