Düzce ili bal arısı (Apis mellifera L.) biyolojik çeşitliliğinin genetik ve morfometrik yöntemler ile araştırılması
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Tarih
2021
Yazarlar
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Cilt Başlığı
Yayıncı
Düzce Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada Batı Karadeniz'de orman gülü, kestane florası ve Yığılca bal arısı ekotipi ile arıcılıkta öne çıkan Düzce ili bal arısı biyolojik çeşitliliğinin morfometrik ve genetik yöntemler ile tanımlanması amaçlanmıştır. Bu amaç için Düzce ili merkez ve ilçelerini temsil edecek şekilde 24 arılıktan, 72 koloniden toplam 1440 işçi arı örneği toplandı. Örneklerin sağ ön kanatlarında landmark temelli 31 morfometrik karakterin yanı sıra dil, kanat ve bacak segmentlerinin uzunlukları ölçüldü. Morfometrik karakterlerin yanı sıra örnekler mitokondriyal genomda 5 gen bölgesi (CoxI- CoxII, 16srDNA, ND5, Cytb ve CoxI)'nin 10 farklı restriksiyon endonükleaz enzimi ile kesim profili bakımından PCR-RFLP yöntemi ve DNA dizi analizi yapılarak ile değerlendirildi. Morfometrik karakterlerin ölçüm sonuçları Diskriminant Fonksiyon Analizi (DFA) ile karşılaştırıldığında Merkez, Akçakoca ve Cumayeri ilçeleri koordinat düzlemi üzerinde birbirlerinden ve diğer ilçelerden belirgin olarak ayrıldı. Gümüşova-Çilimli ve Kaynaşlı-Yığılca ilçelerine ait grup merkezlerinin ikili gruplar şeklinde üst üste çakıştığı ve Gölyaka İlçesi ile yakın kümelendikleri gözlendi. Koloni ortalamaları baz alınarak popülasyonlar arasındaki mahalanobis uzaklıklarına göre oluşturulan UPGMA dendogramında; Akçakoca, Yığılca, Merkez, Çilimli ve Gümüşova birlikte gruplanırken, Cumayeri, Gölyaka ve Kaynaşlı ise birlikte başka bir küme oluşturdu. Örnekleme yapılan her koloniden bir işçi arı örneği alınarak sırasıyla CoxI- CoxII intergenik gen bölgesi DraI, XbaI ve HinfI; CoxI geni SspI, XhoI, DraI ve HinfI; 16srDNA geni SspI, SwaI, DraI, EcoRI ve HincII; ND5 geni HincII, DraI ve SspI; Cyb geni ise BglII, ClaI, HinfI ve DraI resitriksiyon endonükleaz enzimleri ile kesildi. CoxI- CoxII gen bölgesi XbaI ve HinfI kesimlerinde iki farklı mitotip oluşurken, DraI kesiminde üç farklı mitotip oluştu. 16srDNA/SspI kesiminde 2 mitotip oluşmuş, tip 2 yalnızca Cumayeri, Gümüşova, Kaynaşlı ve Merkezde birer örnekte görülürken 16srDNA/HincII kesiminde ise tip 2 yalnızca Yığılca'nın iki örneğinde görüldü. Cytb/DraI kesiminde ise iki farklı mitotip oluştu. Tip 2 profili Gümüşova'nın 6 örneğinde ve Merkez'in 2 örneğinde görüldü. CoxI ve ND5 genlerinin restriksiyon endonükleaz enzimleri ile kesimi sonucu hiçbir farklılık gözlenmedi. Kesim sonuçlarına göre farklı çıkan örneklerin DNA dizi analizleri yaptırıldı. Bu çalışmanın sonuçları Düzce ilinde yaygın görülen mitokondriyal DNA tipinin Anadolu bal arısı (A. m. anatoliaca) mitotipi ile uyumlu olduğunu; ancak Düzce ili arı biyoçeşitliliğinin ana arı ticareti ve göçer arıcılık faaliyetlerinden etkilendiğini göstermiştir.
In this study, the honey bee biodiversity of Düzce province, which stands out in beekeeping with Rhododendron and chestnut flora and Yığılca ecotype in the Western Black Sea Region was studied by using morphometric and genetic methods. For this purpose, a totally 1440 worker bee samples were collected from 24 apiaries, 72 colonies representing all districts of Düzce city. The lengths of the proboscis, wing and leg segments were measured as well as 31 morphometric characters of the right fore wings of the worker honey bee samples. In addition to these morphometric characters, 5 gene regions (CoxI- CoxII, 16srDNA, ND5, Cytb and CoxI) in mtDNA were evaluated with 10 different restriction endonuclease enzymes based on the PCR-RFLP method and by performing DNA sequence analysis. According to Discriminant Function Analysis (DFA) of morphometric data, Merkez, Akçakoca and Cumayeri districts were clearly separated from each other and other districts on the two dimensional plane. It was observed that the group centers of Gümüşova-Çilimli and Kaynaşlı-Yığılca districts overlapped in pairs and clustered closely with Gölyaka District. In the UPGMA dendrogram created according to the mahalanobis distances between the populations based on the colony averages; Akçakoca, Yığılca, Merkez, Çilimli and Gümüşova grouped together, while Cumayeri, Gölyaka and Kaynaşlı formed a group together. A worker bee sample from each colony was analysed regarding to the CoxI-CoxII intergenic region with DraI, XbaI and HınfI; CoxI gene with SspI, XhoI, DraI; 16srDNA gene with SspI, SwaI, DraI, EcoRI and HincII; the ND5 gene with HincII, DraI and SspI; Cytb gene with BglII, ClaI, HınfI and DraI restriction endonuclease enzymes. As a result of the cleavage of the restriction endonuclease enzymes, two different mitotypes were formed in the CoxI-CoxII /XbaI, HınfI, while three different mitotypes were observed in CoxI-CoxII / DraI. Two types were formed in the 16srDNA/SspI segment, type 2 was only seen in one sample from each of Cumayeri, Gümüşova, Kaynaşlı and Merkez districts, while type 2 was seen only in two samples of Yığılca in the 16srDNA/HincII segment. Two different mitotypes were formed in the Cytb/DraI segment. Type 2 profile was seen in 6 samples of Gümüşova and 2 samples of Merkez. No difference was observed as a result of cleavage of CoxI and ND5 genes with restriction endonuclease enzymes. DNA sequence analyzes were applied on the samples which were different according to the PCR/RFLP results. The results showed that mitochondrial DNA type common in Düzce province was compatible with the Anatolian honey bee (A. m. anatoliaca) mitotype; however, honey bee biodiversity of Düzce province was affected by the queen bee trade and migratory beekeeping activities.
In this study, the honey bee biodiversity of Düzce province, which stands out in beekeeping with Rhododendron and chestnut flora and Yığılca ecotype in the Western Black Sea Region was studied by using morphometric and genetic methods. For this purpose, a totally 1440 worker bee samples were collected from 24 apiaries, 72 colonies representing all districts of Düzce city. The lengths of the proboscis, wing and leg segments were measured as well as 31 morphometric characters of the right fore wings of the worker honey bee samples. In addition to these morphometric characters, 5 gene regions (CoxI- CoxII, 16srDNA, ND5, Cytb and CoxI) in mtDNA were evaluated with 10 different restriction endonuclease enzymes based on the PCR-RFLP method and by performing DNA sequence analysis. According to Discriminant Function Analysis (DFA) of morphometric data, Merkez, Akçakoca and Cumayeri districts were clearly separated from each other and other districts on the two dimensional plane. It was observed that the group centers of Gümüşova-Çilimli and Kaynaşlı-Yığılca districts overlapped in pairs and clustered closely with Gölyaka District. In the UPGMA dendrogram created according to the mahalanobis distances between the populations based on the colony averages; Akçakoca, Yığılca, Merkez, Çilimli and Gümüşova grouped together, while Cumayeri, Gölyaka and Kaynaşlı formed a group together. A worker bee sample from each colony was analysed regarding to the CoxI-CoxII intergenic region with DraI, XbaI and HınfI; CoxI gene with SspI, XhoI, DraI; 16srDNA gene with SspI, SwaI, DraI, EcoRI and HincII; the ND5 gene with HincII, DraI and SspI; Cytb gene with BglII, ClaI, HınfI and DraI restriction endonuclease enzymes. As a result of the cleavage of the restriction endonuclease enzymes, two different mitotypes were formed in the CoxI-CoxII /XbaI, HınfI, while three different mitotypes were observed in CoxI-CoxII / DraI. Two types were formed in the 16srDNA/SspI segment, type 2 was only seen in one sample from each of Cumayeri, Gümüşova, Kaynaşlı and Merkez districts, while type 2 was seen only in two samples of Yığılca in the 16srDNA/HincII segment. Two different mitotypes were formed in the Cytb/DraI segment. Type 2 profile was seen in 6 samples of Gümüşova and 2 samples of Merkez. No difference was observed as a result of cleavage of CoxI and ND5 genes with restriction endonuclease enzymes. DNA sequence analyzes were applied on the samples which were different according to the PCR/RFLP results. The results showed that mitochondrial DNA type common in Düzce province was compatible with the Anatolian honey bee (A. m. anatoliaca) mitotype; however, honey bee biodiversity of Düzce province was affected by the queen bee trade and migratory beekeeping activities.
Açıklama
Anahtar Kelimeler
Biyoloji, Biology, Genetik, Genetics, Zooloji, Zoology, Apis mellifera, Apis mellifera, Dizi analizi, Sequence analysis, Morfometri, Morphometry
