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Isolation and Characterization of Three New Promoters from Gossypium hirsutum that Show High Activity in Reproductive Tissues

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Abstract

Engineering of plant protection requires well-characterized tissue-specific promoters for the targeted expression of insecticidal resistance genes. Herein, we describe the isolation of five different fragments of promoters of three distinct flower-specific cotton (Gossypium hirsutum) genes. Expression analyses of the three genes GhPME-like1, GhβGal-like1 and GhPL-like1 revealed that they are expressed highly in flowers buds ranging from 4 to 12 mm in size. Several putative regulatory cis-elements were identified in the promoter regions, including elements involved in the control of tissue-specific gene expression in pollen grains and fruits. In vivo analyses of these promoters were performed using the heterologous plant system Arabidopsis thaliana by fusing them with the gene uidA (GUS). GUS staining in Arabidopsis tissues revealed that their expression was restricted to anthers, with the majority of expression in pollen grains and in the upper portion of the carpels and siliques. A comparison between a CaMV35S::GUS constitutive promoter and the promoters isolated in this study revealed that the cotton promoters were more active and were specific to flowers and fruits, which are organs that are preferentially attacked by important pest insects such as the boll weevil (Anthonomus grandis). The activity of the promoters was also confirmed using transient expression assays in flower buds of G. hirsutum. The promoters of GhPME-like1, GhβGal-like1 and GhPL-like1 are specific to reproductive tissues and could represent important biotechnological tools for controlling insect pests, in particular the cotton boll weevil, which attacks floral and fruit tissues.

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Acknowledgments

We thank the researchers, Fernando Campos de Assis Fonseca and Raquel Sampaio, from Embrapa Genetic Resources and Biotechnology technical assistance with biolistic experiment in cotton. This work was part of S.A’s PhD research in Genetics, at the Department of Genetics of the Universidade Federal do Rio de Janeiro (UFRJ), and was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; M. Alves-Ferreira: # 306025/2010-8) and Fundação de Amparo à Pesquisa do Rio de Janeiro (FAPERJ).

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Authors and Affiliations

  1. Department of Genetics, Federal University of Rio de Janeiro (UFRJ), Av. Prof. Rodolpho Paulo Rocco, s/n - Prédio do CCS Instituto de Biologia, 2° andar, sala A2-93, 219410-970, Rio de Janeiro, RJ, Brazil

    Sinara Artico, Julia Lambret-Frotté, Sarah Muniz Nardeli & Marcio Alves-Ferreira

  2. Parque Estação Biológica (PqEB), Embrapa Genetic Resources and Biotechnology, Av. W5 Norte (final), Caixa Postal 02372, CEP 70770-900, Brasília, DF, Brazil

    Osmundo Brilhante Oliveira-Neto & Maria Fátima Grossi-de-Sa

  3. Department of Biochemistry and Molecular Biology, School of Medicine, FACIPLAC, SIGA AE2, Brasília, 72460-000, DF, Brazil

    Osmundo Brilhante Oliveira-Neto

  4. University Catholic of Brasilia, Brasília, DF, Brazil

    Maria Fátima Grossi-de-Sa

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  1. Sinara Artico
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Correspondence to Marcio Alves-Ferreira.

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Artico, S., Lambret-Frotté, J., Nardeli, S.M. et al. Isolation and Characterization of Three New Promoters from Gossypium hirsutum that Show High Activity in Reproductive Tissues. Plant Mol Biol Rep 32, 630–643 (2014). https://doi.org/10.1007/s11105-013-0674-0

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