Türk kara sinek (Musca domestica L.) soylarında malathion direncinin genetik temeli ve MdαE7 allellerinin dünya üzerindeki dağılımı

Organophosphate (OP) insecticides (parathion / diazinon) resistance in house fly (Musca domestica L.) is associated with the change in carboxylesterase activity. The product of aE7 gene, which is a member of oc-esterase gene cluster, is probably playing a role in detoxyfication of the xenobiotic esters. In diazinon resistant Lucilia cuprina (blowfly) and diazinon / parathion resistant M.domestica species Gly137 to Asp substitution was found in the active center of the product of aE7. This enhance the ability to hydrolyse the OP in so doing the enzyme compromise its ability to hydrolyse carboxylesters "The mutant ali-esterase hypothesis" (Oppenhorth and Asperen, 1960). In malathion (an OP) resistant L. cuprina strains at the position of Trp251 to Leu substitution was found in the product of aE7, also a high malathion 111carboxylesterase activity (MCE) was reported in these strains (Newcomb et al, 1997). In our research, we have isolated, cloned and sequenced the MdaE7 on four different house fly strains: Ankara, Adana and Ceyhanl, 2 (high doses of malathion were applied in laboratory environment for a long time) and Kirazli strain (no insecticide treatment was performed on this strain in the laboratory). Trp251 to Ser substitutions was found in the product of aE7 in all of these strains. In addition, the MCE, glutathion-S-transferase (GST), acetylcholinesterase (AChE), and general esterase activities were tested in these five strains. In all of these tests WHO was used as a standard sensitive control. A high level MCE and GST activities were observed while lower level of general esterase activities was detected in the tested strains. Moreover, a higher percent remaining activities in AChE than WHO susceptible strain were observed in all malathion resistant strains. In order to understand the allelic diversity of the MdaE7 at a worldwide base, this gene was isolated, cloned and sequenced (from the beginning of exon 2 to the end of exon 6) from eleven house fly strains from different localities (Africa, Asia, America and Middle East). It was found out that diazinon resistant type allele was in high frequency around the world, the malathion type resistant alleles seem to be located at only specific regions.Kara sinek (Musca domestica) soylarında organofosfatlı (paration / diazinon) insektisitlere karşı direnç, karboksilesteraz aktivitesindeki değişikliklere bağlıdır, a- esteraz gen ailesinin bir üyesi olan aE7 geninin ürünü, olasılıkla ksenobiotik esterlerin detoksifikasy onunda rol oynamaktadır. Paration / diazinon' a dirençli M. domestica soylarında aE7 geninin ürününün aktif merkezinde Gly137'den Asp'a değiştiği tespit edilmiştir. Bu da enzimin organofosfat insektisitlerini hidroliz edebilme yeteneğini arttırmakta, bunu yaparken de enzimin karboksilesterleri hidroliz edebilme yeteneğini azaltmaktadır "mutant ali-esteraz hipotezi" (Oppenhorth ve Asperen, 1960). Malationa dirençli L. cuprina soylarında aE7 geninin ürününde Trp251 'den Leu'ya değişiklik ve aynı zamanda bu soylarda yüksek malathionkarboksilesteraz aktivitesi bulunmuştur (Newcomb ve ark., 1997). Bu çalışmada laboratuvar ortamında uzun süre yüksek dozlarda malathion uygulanmış olan dört farklı kara sinek soyunda (Ankara, Adana ve Ceyhanl, 2 ) ve laboratuvar ortamında herhangi bir insektisit uygulanmayan Kirazlı soyundan MdccE7 geni izole edilmiş, klonlanmış ve baz dizi analizleri yapılmıştır. Bütün bu soylarda aE7 geninin ürününde Tıp251 'den Ser'e değişiklik bulunmuştur. Bununla birlikte, yukarıda belirtilen beş stokta malathion karboksilesteraz (MKE), glutathion-S-transferaz (GST), asetilkolinesteraz (AKE), ve genel esteraz aktiviteleri test edilmiştir. Bütün bu biyokimyasal testlerde WHO soyu duyarlı standart kontrol olarak kullanılmıştır. Yüksek malathion karboksilesteraz ve glutathion-S-transferaz aktiviteleri gözlenmiş olmasına karşın, düşük seviyede genel esteraz aktivitesi bulunmuştur. Bunlara ek olarak, tüm malathiona dirençli soylarda asetilkolinesteraz enzimi için WHO 'a göre yüksek oranda % kalan aktivite tespit edilmiştir. MdaE7 allellerinin dünya üzerindeki dağılımım belirlemek amacıyla, farklı bölgelerden (Afrika, Asya, Amerika ve Orta Doğu) değişik 1 1 kara sinek soyundan MdaEl geni izole edilmiş, klonlanmış ve baz dizi analizleri (ekzon2'nin başından ekzonö'nın sonuna kadar) yapılmıştır. Diazinona direnç sağlayan allelin dünya üzerinde yüksek frekansa sahip olduğu, buna karşılık malathiona direnç sağlayan allelin sadece belirli bölgelerde yoğunlaştığı görülmüştür.Ph.D. - Doctoral Progra

Genetic diversity of natural cyclamen alpinum populations

The focus of this study was Cyclamen alpinum (formerly C. trochopteranthum). Habitat fragmentation, environmental degradation, and overharvesting of tubers have exerted pressure on native populations of this valuable ornamental species. Although the entire Cyclamen genus is in the Convention on International Trade in Endangered Species of Wild Fauna and Flora-Appendix II (CITES II), no species has yet been red-listed. Estimating the level and distribution of genetic variation in populations of rare and endemic species is important for conserving genetic diversity within a species in the context of well-developed conservation strategies. Currently, DNA markers are the most effective means used to infer genetic variation at the molecular level in conservation genetics. In this study, random amplified polymorphic DNA (RAPD) analysis was employed to assess the genetic diversity within and among 6 natural C. alpinum populations in the south and southwest of Turkey. A total of 190 loci were determined by using 15 polymorphic primers. Total genetic variation (HT) was 0.27 ± 0.02. A high proportion of this variation, 0.16 ± 0.01 (59.26%), was due to withinpopulation genetic variation (HS). The genetic differentiation coefficient (GST) was 0.41, and the level of gene flow (Nm) within a generation among the 6 populations studied was 0.73. As a result of these findings, we propose in situ combined withex situ conservation of all C. alpinum populations. In addition, our results support prior recommendations to add C. alpinum to the International Union for Conservation of Nature and Natural Resources (IUCN) Red List under the critically endangered (CR) category. © TÜBİTAK