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Allelic Variability in the Intronic Region of the Fibroin Heavy-Chain Gene in Silkworm Bombyx mori L. Strains of Brazilian Germplasm Bank

  • Systematics, Morphology and Physiology
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Abstract

The domesticated silkworm Bombyx mori L. is currently found only in germplasm banks. Therefore, characterization and conservation of this genetic resource is crucial. Based on previous studies that revealed nucleotide differences in silkworm strains, the intron of the fibroin heavy chain gene (H-fib) can be used for molecular silkworm characterization. The H-fib gene has two exons and a unique intron, and encodes the principal component of the silk fiber, the fibroin heavy chain. Therefore, this study aimed to identify the genetic variability of the unique intron of H-fib gene of 20 silkworm strains maintained at the Universidade Estadual de Maringá Brazilian Germplasm Bank (UBGB) by conformation-sensitive gel electrophoresis (CSGE) and nucleotide sequencing. Genomic DNA extracted from silkworm moths was PCR amplified. CSGE revealed that most of the analyzed silkworm strains had only homoduplex molecules. However, DNA from the Japanese strains B106, B82, and M12-2 had two extra DNA fragments produced by heteroduplex molecules, revealing variation between alleles. Sequencing of the H-fib intron was used to confirm the variation previously detected by CSGE and detected a significant polymorphism characterized by a 17-base pair (bp) deletion, a 2-bp insertion, and eight nucleotide substitutions. Although genetic and allelic variability was detected in some silkworm strains, the intron of the H-fib gene revealed not to be the best molecular marker for the characterization of B. mori strains from UBGB.

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Acknowledgments

This research was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Financiadora de Estudos e Projetos (FINEP)/Fundação Araucária and Secretaria de Estado da Ciência, Tecnologia e Ensino Superior – FUNDO PARANÁ. Thanks also to CAPES for the post-doctorate fellowship to TS Bignotto. Authors gratefully thank all staff members of Laboratório de Organização Functional do Núcleo (LORF), Complexo de Centrais de Apoio à Pesquisa (COMCAP) and Seda y Fibras S.R.L. We also thank anonymous reviewers for their comments and valuable suggestions.

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Authors and Affiliations

  1. Depto de Biotecnologia, Biologia Celular e Genética, Univ Estadual de Maringá, Av. Colombo, 5790, Bloco B36, 87020-900, Maringá, PR, Brasil

    T S Bignotto, R E F Munhoz, N C Pereira, R Bespalhuk, C R N Saez, V A Fassina & M A Fernandez

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  1. T S Bignotto
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  2. R E F Munhoz
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  3. N C Pereira
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  4. R Bespalhuk
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  5. C R N Saez
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  6. V A Fassina
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  7. M A Fernandez
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Correspondence to T S Bignotto.

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Edited by Anne-Nathalie Volkoff – INRA/Univ of Montpellier

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Bignotto, T.S., Munhoz, R.E.F., Pereira, N.C. et al. Allelic Variability in the Intronic Region of the Fibroin Heavy-Chain Gene in Silkworm Bombyx mori L. Strains of Brazilian Germplasm Bank. Neotrop Entomol 43, 252–259 (2014). https://doi.org/10.1007/s13744-014-0205-1

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