RT-PCR Analysis of Caucasian and Mugla Honey Bees by SNP Markers of Chalkbrood Disease

Abstract

Two different honey bee subspecies’ genotypes obtained from Ordu, Apiculture Research Institute were analyzed by SNP markers using Real-Time PCR-HRM. Genomic DNA samples analysed with 10 SNP primers those were used for identification of chalkbrood disease resistance genes and two SNP primers those were obtained from honey bee genom sequencing. Result of SNP analyses, four primers (AMB-00858574, AMB-01151447, AMB00631190, AMB-00686140) amplified in Caucasian honey bee and six (AMB-00858574, AMB-00612262, AMB-01151447, AMB-00631190, AMB-00674355, AMB-00686140) primers amplified in Mugla honey bee. Four amplicons are similar for this subspecies. Result of electrophoresis analyses, five primers (AMB00858574, AMB-00612262, AMB-01151447, AMB-00631190, AMB-00686140) form a band in Caucasian honey bee, seven amplicons (AMB-00858574, AMB00612262, AMB-01151447, AMB-00631190, AMB-00902548, AMB-00674355, AMB-00686140) form a band in Mugla honey bee and four amplicons (AMB00858574, AMB-01151447, AMB-00631190, AMB-00686140) similar for this subspecies. As a result of this study, in Caucasian and Muğla honeybee for identification of disease resistance and evaluability as a identification key for this subspecies was emerged the capacity of association of single nucleotide polymorphisms to resistance to chalkbrood in two important honeybee genotypes in country of Apis mellifera using RT-PCR for HRM analysis.

Keywords

• Honey Bee
• SNP
• Mugla bee
• Caucasian bee
• Real-Time PCR
• HRM
• chalkbrood disease

Kafkas ve Muğla Bal Arılarının Tebeşir Hastalığının SNP Belirteçleri ile RT-PCR Analizi

Öz

Ordu, Arıcılık Araştırma Enstitüsü'nden sağlanan iki farklı bal arısı alt türüne ait genotipler Real-Time PCR-HRM kullanılarak SNP markörleri ile analiz edilmiştir. Kireç hastalığı direnç genlerinin tanımlanmasında kullanılan 10 SNP primeri ve bal arısı genom dizilemesinden elde edilen iki SNP primeri ile analiz edilen genomik DNA örneklerinde; Kafkas bal arısında amplifiye edilmiş dört primer (AMB-00858574, AMB-01151447, AMB 00631190, AMB-00686140) ve altı (AMB-00858574, AMB-00612262, AMB-01151447, AMB-00631190, AMB-00674355, AMB-00686140) Muğla bal arısında amplifiye edilmiştir. Bu alt tür için dört amplikon benzer olup elektroforez analizleri sonucunda, Kafkas bal arısında beş primer (AMB 00858574, AMB-00612262, AMB-01151447, AMB-00631190, AMB-00686140), yedi amplikon (AMB-00858574, AMB 00612262, AMB-01151447, AMB -00631190, AMB-00902548, AMB-00674355, AMB-00686140) Muğla bal arısında dört amplikon (AMB 00858574, AMB-01151447, AMB-00631190, AMB-00686140) bir bant oluşturmuştur. Bu çalışma sonucunda, Kafkas ve Muğla bal arısı genotiplerinde hastalık direncinin belirlenmesi ve bu alt tür için bir tanımlama anahtarı olarak değerlendirilebileceği, HRM analizi için RT-PCR kullanarak Apis mellifera'nın önemli iki bal arısı genotipinde tek nükleotid polimorfizmlerinin kireç direnci ile ilişkilendirme kapasitesi ortaya çıkarılmıştır.

Anahtar Kelimeler

• Bal arısı
• Mugla arısı
• Kafkas arısı
• SNP
• RT-PCR
• HRM
• kireç hastalığı

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