Entomopatojen nematod Heterorhabditis bacteriophora HBH hibrit ırkının in vivo ve in vitro üretim sonrası üreme potansiyeli ve etkinlik farklılıklarının araştırılması
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Date
2021-07-29
Authors
Özbudak, Gizem
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Publisher
Bursa Uludağ Üniversitesi
Abstract
Entomopatojen nematod (EPN) Heterorhabditis bacteriophora HBH ırkı, simbiyotik bakteri olan Photorhabdus ile mutualist bir ilişki içindedir. Bu mutualist ilişki, EPN’ lerin tarımsal açıdan bazı zararlılara karşı, kimyasal insektisitlere alternatif olarak kullanılmasına olanak sağlamaktadır. EPN’ ler in vivo ve in vitro olmak üzere iki ayrı yöntem ile üretilmektedir. In vivo üretim, genellikle laboratuvar çalışmalarında az miktarda üretilmiş olan nematod kültürlerinin devamlılığı ve muhafazası için kullanılan canlı materyal üzerinde bir yöntemdir. Fakat bu yöntem EPN’ lerin ticari amaçla kitlesel üretiminde yeterli olmamaktadır. Kitlesel üretim amacıyla, yapay besiyeri kullanılarak in vitro yöntemiyle EPN üretimi yapılmaktadır. In vivo üretim ile elde edilen infektif juveniller (IJ), üreme potansiyelini tespit etmek amacıyla Galleria mellonella larvalarına karşı uygulanmıştır. Bu larvalar disekte edildiğinde H. bacteriophora HBH ırkına ait hermafrodit birey sayısının (140) 4. günde maksimum seviyede olduğu ve en düşük hermafrodit birey sayısının (95,8) 3. günde elde edildiği tespit edilmiştir. In vitro üretim ile elde edilen H. bacteriophora HBH ırkı, G. mellonella larvalarına karşı uygulandığında ise en yüksek hermafrodit sayısı (86) 6. ve 7. gün tespit edilirken, en düşük hermafrodit sayısı (47,5) 4. gün tespit edilmiştir. Ayrıca, H. bacteriophora HBH ırkı EPN’ lerin etkinliğinin belirlenmesi amacıyla G. mellonella larvaları kullanılmıştır. Bu uygulamada 10 IJ/larva dozunda, in vivo üretim sonucu elde edilen H. bacteriophora HBH ırkının G. Mellonella üzerinde ölüm oranı %100 olurken, in vitro üretim sonucu G. mellonella ölüm oranı %85,6 olmuştur. Özetle, bu tez çalışması in vivo ve in vitro üretim yöntemleri EPN’ lerin etkinlik ve üreme potansiyellerini önemli ölçüde değiştirebileceği tespit edilmiştir.
The strain Heterorhabditis bacteriophora HBH, an entomopathogenic nematode (EPN), is with the symbiotic bacterium Photorhabdus. This mutualistic relationship allows EPNs to be used as an alternative to chemical insecticides against some agricultural pests. EPNs are produced by two different methods, in vivo and in vitro. İn vivo production is a method on living material used for the maintenance and preservation of nematode cultures that are usually produced in small quantities in laboratory studies. However, this method is not sufficient for mass production of EPNs for commercial purposes. For mass production, EPN is produced by in vitro method using artificial media. Infective juveniles of Heterorhabditis bacteriophora HBH obtained by in vivo production were applied against Galleria mellonella larvae to determine their reproductive potential. When these larvae were dissected, it was determined that the number of hermaphrodite individuals belonging to the H. bacteriophora HBH strain was at the maximum level (140) on the 4th day and the lowest number (95,8) of hermaphrodite individuals was obtained on the 3rd day. When H. bacteriophora HBH strain obtained by in vitro production was applied against G. mellonella larvae, the highest hermaphrodite number (86) was detected on the 6th and 7th day, while the lowest number of hermaphrodites (47.5) was determined on the 4th day. In addition, G. mellonella larvae were used to determine the effectiveness of H. bacteriophora HBH strain EPNs. In this application, at a dose of 10 IJ/larvae, the mortality rate of H. bacteriophora HBH strain obtained as a result of in vivo production on G. mellonella was 100%, while the mortality rate of G. mellonella as a result of in vitro production was 85.6%. In summary, at the end of this thesis, it has been determined that in vivo and in vitro production methods can significantly change the efficacy and reproductive potential of EPNs.
The strain Heterorhabditis bacteriophora HBH, an entomopathogenic nematode (EPN), is with the symbiotic bacterium Photorhabdus. This mutualistic relationship allows EPNs to be used as an alternative to chemical insecticides against some agricultural pests. EPNs are produced by two different methods, in vivo and in vitro. İn vivo production is a method on living material used for the maintenance and preservation of nematode cultures that are usually produced in small quantities in laboratory studies. However, this method is not sufficient for mass production of EPNs for commercial purposes. For mass production, EPN is produced by in vitro method using artificial media. Infective juveniles of Heterorhabditis bacteriophora HBH obtained by in vivo production were applied against Galleria mellonella larvae to determine their reproductive potential. When these larvae were dissected, it was determined that the number of hermaphrodite individuals belonging to the H. bacteriophora HBH strain was at the maximum level (140) on the 4th day and the lowest number (95,8) of hermaphrodite individuals was obtained on the 3rd day. When H. bacteriophora HBH strain obtained by in vitro production was applied against G. mellonella larvae, the highest hermaphrodite number (86) was detected on the 6th and 7th day, while the lowest number of hermaphrodites (47.5) was determined on the 4th day. In addition, G. mellonella larvae were used to determine the effectiveness of H. bacteriophora HBH strain EPNs. In this application, at a dose of 10 IJ/larvae, the mortality rate of H. bacteriophora HBH strain obtained as a result of in vivo production on G. mellonella was 100%, while the mortality rate of G. mellonella as a result of in vitro production was 85.6%. In summary, at the end of this thesis, it has been determined that in vivo and in vitro production methods can significantly change the efficacy and reproductive potential of EPNs.
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Keywords
In vivo üretim, In vitro üretim, Entomopatojen nematod, Heterorhabditis bacteriophora HBH, In vivo, In vitro, Entomopathogenic nematodes
Citation
Özbudak, G. (2021). Entomopatojen nematod Heterorhabditis bacteriophora HBH hibrit ırkının in vivo ve in vitro üretim sonrası üreme potansiyeli ve etkinlik farklılıklarının araştırılması. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Fen Bilimleri Enstitüsü.