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Expressed sequence-tag analysis of ovaries of Brachiaria brizantha reveals genes associated with the early steps of embryo sac differentiation of apomictic plants

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Abstract

In apomixis, asexual mode of plant reproduction through seeds, an unreduced megagametophyte is formed due to circumvented or altered meiosis. The embryo develops autonomously from the unreduced egg cell, independently of fertilization. Brachiaria is a genus of tropical forage grasses that reproduces sexually or by apomixis. A limited number of studies have reported the sequencing of apomixis-related genes and a few Brachiaria sequences have been deposited at genebank databases. This work shows sequencing and expression analyses of expressed sequence-tags (ESTs) of Brachiaria genus and points to transcripts from ovaries with preferential expression at megasporogenesis in apomictic plants. From the 11 differentially expressed sequences from immature ovaries of sexual and apomictic Brachiaria brizantha obtained from macroarray analysis, 9 were preferentially detected in ovaries of apomicts, as confirmed by RT-qPCR. A putative involvement in early steps of Panicum-type embryo sac differentiation of four sequences from B. brizantha ovaries: BbrizHelic, BbrizRan, BbrizSec13 and BbrizSti1 is suggested. Two of these, BbrizSti1 and BbrizHelic, with similarity to a gene coding to stress induced protein and a helicase, respectively, are preferentially expressed in the early stages of apomictic ovaries development, especially in the nucellus, in a stage previous to the differentiation of aposporous initials, as verified by in situ hybridization.

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Acknowledgments

The authors gratefully acknowledge Samara Kalaoun, from UFRJ, for help with RT-qPCR experiments; Dr. Milton Moraes, from Fiocruz, for support material and equipment for RT-qPCR. From Embrapa, they thank Rafael W. de Souza, for dissecting ovaries; Dr. Luiz JCB. Carvalho and Dr. Marco V. Agostin for technical support and analyses of macroarray experiments; and Dr Julio CM. Rodrigues for comments on the manuscript. This work is part of EDS’ PhD thesis from Pós-Graduação em Biotecnologia Vegetal, UFRJ, Brazil, with a partial fellowship from CAPES, CNPq and Embrapa. This work was partially supported by CNPq (478356/2006-4 – VTCC), Embrapa (02.05.1.02.00- VTCC) and Fundação de Amparo à Pesquisa do Rio de Janeiro (FAPERJ; E-26/102.861/2008 MA-F).

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  1. Érica Duarte Silveira

    Present address: Facultad de Ciencias Agrarias, Universidad Catolica Argentina, Cap. Gral. Ramón Freire 183, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina

Authors and Affiliations

  1. Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica, PqEB, Av. W5 Norte (final) Caixa Postal 02372, 70770917, Brasília DF, Brazil

    Érica Duarte Silveira, Larissa Arrais Guimarães, Diva Maria de Alencar Dusi, Felipe Rodrigues da Silva, Natália Florencio Martins, Marcos Mota do Carmo Costa & Vera Tavares de Campos Carneiro

  2. Department of Genetics, Federal University of Rio de Janeiro, Av. Prof. Rodolpho Paulo Rocco, s/n Prédio do CCS Instituto de Biologia, 2 andar, 21941-970, Rio de Janeiro, RJ, Brazil

    Érica Duarte Silveira & Márcio Alves-Ferreira

  3. Department of Cell Biology, University of Brasília, UnB, Campus Universitário Darcy Ribeiro, S/N-Asa Norte, 70919- 970, Brasília DF, Brazil

    Larissa Arrais Guimarães

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  1. Érica Duarte Silveira
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Communicated by K. Wang.

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299_2011_1175_MOESM1_ESM.doc

Supplementary Table 1 Expressed sequence tags (ESTs) derived from libraries of ovaries at megasporogenesis of apomictic (B030_IeII), and sexual (B105_IeII) plants of Brachiaria brizantha analysed by macroarray. Sequence search results (best matches) of ESTs with significant similarities by BLASTN and BLASTX to sequences already related to important processes of gamete differentiation and ESTs with no hits are listed (DOC 314 kb)

299_2011_1175_MOESM2_ESM.doc

Supplementary Table 2 Oligonucleotides designed for RT-qPCR analyses of differential ESTs in sexual and apomictic Brachiaria brizantha (DOC 34 kb)

299_2011_1175_MOESM3_ESM.doc

Supplementary Table 3 Oligonucleotides designed for RACE of the BbrizSti1 and BbrizHelic ESTs in sexual and apomictic Brachiaria brizantha (DOC 31 kb)

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Silveira, É.D., Guimarães, L.A., de Alencar Dusi, D.M. et al. Expressed sequence-tag analysis of ovaries of Brachiaria brizantha reveals genes associated with the early steps of embryo sac differentiation of apomictic plants. Plant Cell Rep 31, 403–416 (2012). https://doi.org/10.1007/s00299-011-1175-y

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