Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA

The <i>complementary sex determiner</i> (<i>csd</i>) gene plays an essential role in the sex determination of <i>Apis mellifera</i> L. Females develop only if fertilized eggs have functional heterozygous genotypes at this gene whereas males, being haploids, are he...

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Main Authors: Samuele Bovo, Anisa Ribani, Valerio Joe Utzeri, Valeria Taurisano, Giuseppina Schiavo, Matteo Bolner, Luca Fontanesi
Format: Article
Language:English
Published: MDPI AG 2021-09-01
Series:Insects
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Online Access:https://www.mdpi.com/2075-4450/12/10/868
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author Samuele Bovo
Anisa Ribani
Valerio Joe Utzeri
Valeria Taurisano
Giuseppina Schiavo
Matteo Bolner
Luca Fontanesi
author_facet Samuele Bovo
Anisa Ribani
Valerio Joe Utzeri
Valeria Taurisano
Giuseppina Schiavo
Matteo Bolner
Luca Fontanesi
author_sort Samuele Bovo
collection DOAJ
description The <i>complementary sex determiner</i> (<i>csd</i>) gene plays an essential role in the sex determination of <i>Apis mellifera</i> L. Females develop only if fertilized eggs have functional heterozygous genotypes at this gene whereas males, being haploids, are hemizygous. Two identical <i>csd</i> alleles produce non vital males. In light of the recent decline in honey bee populations, it is therefore important to monitor the allele variability at this gene. In this study, we tested the application of next generation semiconductor-based sequencing technology (Ion Torrent) coupled with environmental honey DNA as a source of honey bee genome information to retrieve massive sequencing data for the analysis of variability at the hypervariable region (HVR) of the <i>csd</i> gene. DNA was extracted from 12 honey samples collected from honeycombs directly retrieved from 12 different colonies. A specifically designed bioinformatic pipeline, applied to analyze a total of about 1.5 million reads, identified a total of 160 different <i>csd</i> alleles, 55% of which were novel. The average number of alleles per sample was compatible with the number of expected patrilines per colony, according to the mating behavior of the queens. Allele diversity at the <i>csd</i> could also provide information useful to reconstruct the history of the honey.
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spelling doaj.art-729e34e5ec33463fb76c6c68643cb7132023-11-22T18:38:47ZengMDPI AGInsects2075-44502021-09-01121086810.3390/insects12100868Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNASamuele Bovo0Anisa Ribani1Valerio Joe Utzeri2Valeria Taurisano3Giuseppina Schiavo4Matteo Bolner5Luca Fontanesi6Department of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyDepartment of Agricultural and Food Sciences, University of Bologna, Viale Giuseppe Fanin 46, 40127 Bologna, ItalyThe <i>complementary sex determiner</i> (<i>csd</i>) gene plays an essential role in the sex determination of <i>Apis mellifera</i> L. Females develop only if fertilized eggs have functional heterozygous genotypes at this gene whereas males, being haploids, are hemizygous. Two identical <i>csd</i> alleles produce non vital males. In light of the recent decline in honey bee populations, it is therefore important to monitor the allele variability at this gene. In this study, we tested the application of next generation semiconductor-based sequencing technology (Ion Torrent) coupled with environmental honey DNA as a source of honey bee genome information to retrieve massive sequencing data for the analysis of variability at the hypervariable region (HVR) of the <i>csd</i> gene. DNA was extracted from 12 honey samples collected from honeycombs directly retrieved from 12 different colonies. A specifically designed bioinformatic pipeline, applied to analyze a total of about 1.5 million reads, identified a total of 160 different <i>csd</i> alleles, 55% of which were novel. The average number of alleles per sample was compatible with the number of expected patrilines per colony, according to the mating behavior of the queens. Allele diversity at the <i>csd</i> could also provide information useful to reconstruct the history of the honey.https://www.mdpi.com/2075-4450/12/10/868environmental DNAgenetic diversityhoney beeinbreedingpolymorphismpopulation genomics
spellingShingle Samuele Bovo
Anisa Ribani
Valerio Joe Utzeri
Valeria Taurisano
Giuseppina Schiavo
Matteo Bolner
Luca Fontanesi
Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
Insects
environmental DNA
genetic diversity
honey bee
inbreeding
polymorphism
population genomics
title Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
title_full Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
title_fullStr Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
title_full_unstemmed Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
title_short Application of Next Generation Semiconductor-Based Sequencing for the Identification of <i>Apis mellifera Complementary Sex Determiner</i> (<i>csd</i>) Alleles from Honey DNA
title_sort application of next generation semiconductor based sequencing for the identification of i apis mellifera complementary sex determiner i i csd i alleles from honey dna
topic environmental DNA
genetic diversity
honey bee
inbreeding
polymorphism
population genomics
url https://www.mdpi.com/2075-4450/12/10/868
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