Abstract
Main conclusion
An exonuclease V homologue from apomictic Brachiaria brizantha is expressed and localized in nucellar cells at key moments when these cells differentiate to give rise to unreduced gametophytes.
Abstract
Brachiaria is a genus of forage grasses with economical and agricultural importance to Brazil. Brachiaria reproduces by aposporic apomixis, in which unreduced embryo sacs, derived from nucellar cells, other than the megaspore mother cell (MMC), are formed. The unreduced embryo sacs produce an embryo without fertilization resulting in clones of the mother plant. Comparative gene expression analysis in ovaries of sexual and apomictic Brachiaria spp. revealed a sequence from B. brizantha that showed a distinct pattern of expression in ovaries of sexual and apomictic plants. In this work, we describe a gene named BbrizExoV with strong identity to exonuclease V (Exo V) genes from other grasses. Sequence analysis in signal prediction tools showed that BbrizExoV might have dual localization, depending on the translation point. A longer form to the nucleus and a shorter form which would be directed to the chloroplast. This is also the case for monocot sequences analyzed from other species. The long form of BbrizExoV protein localizes to the nucleus of onion epidermal cells. Analysis of ExoV proteins from dicot species, with exception of Arabidopsis thaliana ExoVL protein, showed only one localization. Using a template-based AlphaFold 2 modelling approach the structure of BbrizExoV in complex with metal and ssDNA was predicted based on the holo structure of the human counterpart. Features predicted to define ssDNA binding but a lack of sequence specificity are shared between the human enzyme and BbrizExoV. Expression analyses indicated the precise site and timing of transcript accumulation during ovule development, which coincides with the differentiation of nucelar cells to form the typical aposporic four-celled unreduced gametophyte. A putative function for this protein is proposed based on its homology and expression pattern.
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Data availability
The Brachiaria brizantha ExoV sequence analysed in this study was deposited in the NCBI GenBank database under the accession number BankIt2615306 exonuclease OP294995. All data generated or analyzed during this study are included in this published article (and its supplementary information files).
Abbreviations
- AI:
-
Apospore initial cells
- DAPI:
-
4′,6-Diamidine-2′-phenylindole dihydrochloride
- GFP:
-
Green fluorescent protein
- MMC:
-
Megaspore mother cell
- p35S:
-
Constitutive promoter of Cauliflower mosaic virus
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The authors acknowledge the finnancial support of the Brazilian Agricultural Research Corporation-Embrapa, the National Council for Scientific and Technological Development-CNPq and the Coordination for the Improvement of Higher Level Personnel, CAPES-UnB and CNPq for thesis fellowships.
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Dusi, D.M.A., Alves, E.R., Cabral, G.B. et al. An exonuclease V homologue is expressed predominantly during early megasporogenesis in apomictic Brachiaria brizantha. Planta 258, 5 (2023). https://doi.org/10.1007/s00425-023-04162-8
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DOI: https://doi.org/10.1007/s00425-023-04162-8