Bazı fungus ve bakterilerde detoksifikasyon sistemi ve sistem üzerine pestisitlerin etkisinin araştırılması
Küçük Resim Yok
Tarih
1998
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
ÖZET Bu çalışmada Basidiomycetes sınıfına dahil olan Funalia trogii ve Ascomycetes sınıfından Aspergillus niger kullanılmıştır. Araştırmada ayrıca Escherchia coli, Pseudomonas aeruginosa, Pseudomanas putida ve Staphylococcus aureus bakterileri ile de çalışılmıştır. Çalışmada mikroorganizmaların detoksifikasyon sistemi ile ilgili oldukları düşünülen katalaz (CAT), glutatyon redüktaz (GR), glutatyon S- transferaz (GST) süperoksit dismutaz (SOD) ve redükte glutatyon seviyelerinin zamana bağlı değişimleri araştırılmıştır. Ayrıca bu sistem üzerine çeşitli pestisitlerin etkisi de test edilmiştir. Statik ve çalkalamak koşullarda üretilen F. Trogii'nin CAT ve GR aktiviteleri kontrol grubunda üremenin erken fazlarında yüksek iken geç fazlara doğru düşmüştür. GST ve SOD aktiviteleri ile GSH miktarında ise artış saptanmıştır. Çalkalamak koşullarda üretilen F. trogii 'ye insektisit uygulanması katalaz aktivitesini geç fazlara doğru indüklerken, statik koşullarda insektisit uygulanması katalaz aktivitesini üremenin geç fazlarında azaltmıştır. İnsektisit eklenmesi sonucunda her iki koşulda da, statik ve çalkalamak, genelde GR, GST ve SOD enzim aktivitelerinde ve GSH miktarında azalma saptanmıştır. A. niger ile çalkalamak koşullarda insektisit eklenmeyen besiyerinde yürütülen çalışmada katalaz aktivitesi üremenin başlangıcında artmış ancak 4. ve 5. günlerde ise azalmıştır. GST ve GR aktiviteleri de zamana bağlı olarak azalırken, GSH miktarının ise üremenin erken fazlarında düşük olup, 5. günde arrtığı saptanmıştır. SOD aktivitesi ise fazla bir değişim göstermeyip üremenin geç safhalarında artmıştır. İnsektisit uygulaması ise GR aktivitesini azaltırken üremenin 2. ve 5. gününde GST aktivitesini, 4. ve 5. günlerde katalaz aktivitesini, 1. ve 2. günlerde ise SOD aktivitesini artırmıştır. Çalışmada ayrıca bir herbisit olan paraquat'ın F. Trogii'de bu enzimler ve glutatyon üzerine etkisi de test edilmiştir. Üretimin başlangıcında 0.1 ve 1,0 mM paraquat uygulamaları CAT aktivitesini ve GSH miktarım azaltırken, SOD, GR ve GST aktivitelerini ise artırmıştır. Üretim sonrası paraquat uygulaması ise CAT aktivitesini azaltmışta. 0.1 mM'lık uygulama, GR aktivitesini artırmış, 1.0 ve 10.0 mM'lık uygulamalar ise aktiviteyi azaltmıştır. GST aktivitesi ise 6. günde herbisit eklenmesi sonucunda artmıştır. Her üç konsantrasyonda da GSH miktarı azalırken SOD aktivitesi artmıştır. Genel olarak F. trogii ye paraquat uygulamaları SOD aktivitesinin artışına ve CAT aktivitesinin düşmesine neden olmuştur. Bu paraquat uygulaması sonucunda hidrojen peroksit birikimine bağlı olarak diğer organizmalarda da olumsuz etki ortaya çıkabileceğini göstermektedir. Bu yüzden paraquatin dikkatli kullanılması gereklidir. Çalışmamızda ayrıca çeşitli bakterilerin detoksifikasyondan sorumlu enzim aktiviteleri ve redükte glutatyon miktarındaki değişimler de araştırılmıştır. E. coli, P. Putida ve S. aureus' da CAT ve GR aktivitelerinin zamana bağlı dalgalanmalar gösterdiği gözlenmiştir. GST, SOD aktiviteleri ve GSH miktarı ise zamana bağlı olarak azalmıştır. Bu açıdan F. trogii ve bakteriler farklılık göstermektedirler. Bu çalışmada son olarak P. aeruginosa üzerine üremenin başlangıcında ve üreme sonrası iki farklı şekilde eklenen insektisitin etkisi araştırılmıştır. P. aeruginosa 'nın besiyerine üretimin başlangıcında eklenen 5 ve 20 ppm insektisit katalaz aktivitesini azaltırken GR aktivitesini artırmıştır. SOD aktivitesi ise m 4. saatte baskılanırken daha sonraki saatlerde indüklenmiştir. GST aktivitesi genel olarak azalırken, GSH miktarı bu azalmanın tersine artmıştır. Üretimin 8. saatında eklenen 5 ve 20 ppm insektisit, katalaz aktivitesini 30 dakikada artırırken 0, 60 ve 120. dakikalarda düşürmüştür. GR aktivitesi 30. dakikada artmış ancak 0 ve 120. dakikalarda baskılandığı gözlenmiştir. GST aktivitesi 5 ppm'lik uygulamalar sonucunda 0 ve 30. dakikalarda azalmıştır. Yüksek doz uygulama sonucunda (20 ppm) ise 60 ve 120. dakikalarda aktivite artmıştır. Bunların yanısıra GSH miktarı azalırken SOD aktivitesi artmıştır. Sonuç olarak mikroorganizmaların detoksifikasyon sisteminde yer alan bu ajanların zamana bağlı değişimi mikroorganizmadan mikroorganizmaya farklılık göstermektedir. Yine pestisit uygulamalarının etkisi de kullanılan mikroorganizmaya ve üreme fazına bağlı olarak farklılık içermektedir. Çalışmada mikroorganaizmaların detoksifikasyon sistemi ilk kez bu kadar geniş araştırılmış ve ilk kez zamana bağlı değişim ortaya konmaya çalışılmıştır. Anahtar Kelimeler: Süperoksit dismutaz, Katalaz, Glutatyon redüktaz, Glutatyon S- transferaz, Glutatyon, Fungus, Bakteri, Pestisid.
IV ABSTRACT In this study, the detoxification systems and the effects of various pesticides on these systems were investigated using two fungal species, F. trogii and A. niger, and also bacteria E. coli, P. aeruginosa, P. putida and S. aureus. Here, the level of reduced glutathione and various enzymes catalase (CAT), glutathione reductase (GR), glutathione S- transferase (GST) and süperoxide dismutase (SOD), which are known for their detoxification capabilities in microorganisms was tested in a time depending manner for any change in their level and activity. Also, the effect of various pesticides was tested on this system. In the control groups of F. trogii grown in shaking and non-shaking conditions, the activity of CAT and GR in early stage of growth was found to be higher than the late stages and GST, SOD activities and GSH level were seen to be increased. However, while the activity of CAT was found to be increased at the late phase of growth, under shaking incubation conditions, the non-shaking incubation caused a reverse effect, mainly decreasing the activity of CAT. Under both conditions, GR, GST and SOD activities and GSH amount were reduced by the addition of the insecticide. While the CAT activity was increased at the begining of the growth, it was decreased at the 4th and 5th day of growth under shaking conditions in control group of A. niger. Also the activities of GST and GR were decreased in time but the low level of GSH at the begining of growth was seen to be increased at the 5th day of incubation. No change was observed in SOD activity, except at the late phases of growth where it was seen to be increased. The treatment of A. niger cultures with insectiside caused a decrease in GR activity and in increase in the activity of GST, CAT and SOD at the 2nd and 5lh, 4lh and 5th, 1st and 2nd days, respectively. The effect of paraquat, a herbicide, was tested on activity those enzymes and GSH levels of F. trogii. While the CAT activity and the level of GSH of F. trogii at the beginning of incubation was found to be decreased with 0.1 and 1.0 mM concentrations of paraquat, GR, SOD and GST activities were found to be increased. It was found that the CAT activity was decreased, but GR activity was increased with the application of 0.1 mM paraquat to the cultures at late growth phase. But 1.0 and 10.0 mM concentration caused a decrease in GR activity. The activity of GST was increased with the addition of herbicide at the 6th day. The level of GSH was decreased and SOD activity was increase with all three concentrations of herbicide. In general the treatment of F. trogii with paraquat caused an increased in SOD activity but a decrease in CAT activity. The accumulation of H202 after paraquat treatment may cause undesirable effect in other organisms. So, paraquat treatments should be carried out with great caution. Also, in this study, the activity of enzymes responsible for detoxification and the level of GSH, in several bacteria, were tested. There was fluctations in the activity of CAT and GR in E. coli, P. putida, S. aureus throughout the incubation time. The activity of GST and SOD and the level of GSH was seen to be decreased in time J7. trogii and those bacteria were seen to be differently behave in this respect. VI In a part of this study, we have also tested the effect of insectiside, guthion, onp. aeruginosa by the treatment of cultures both at the begining of incubation and at the late growth phases. The effect of insecticide on bacteria was studied by the addition of 5 and 20 ppm guthion into the growth media of P. aeruginosa. These concentrations caused a repression in CAT activity but an induction in GR activity. Also, while SOD activity was repressed at the 4th hour of incubation, an induction of it was observed at the later times of incubation with the application of same amount of insecticide. While the GST activity decreased the level of GSH propontionally increased. At the 8th hour of incubation, 5 and 20 ppm insecticide caused an increase at the 30 min after its addition, but a decrease was observed at the 0, 60 and 120 min of addition. While the activity of GR was increased at 30 minutes of incubations at 0 and 120 minutes of incubation. Its activity was seen to be decreased. The addition of 5 ppm caused a decreased in GST activity at 0 and 30 min, an increase in 20 ppm its activity at 60 and 120 min of incubation. Also, while the GSH level decreased the activity of SOD was observed to be increased. It was concluded that there were fluctations and differences in those systems of detoxification among microorganisms used. The treatment of those microorganisms with pesticides also caused similar changes depending mainly on the growth phase. This is a study in which the detoxification systems of microorganisms is so widely studied for the first time and also an attempt was made to figure out the changes of those agents throughout the incubation period. vıı Key Words: Süperoxide dismutase, Catalase, Glutathione reductase, Glutathione S- transferase, Glutathione, Fungus, Bacterium, Pesticide.
IV ABSTRACT In this study, the detoxification systems and the effects of various pesticides on these systems were investigated using two fungal species, F. trogii and A. niger, and also bacteria E. coli, P. aeruginosa, P. putida and S. aureus. Here, the level of reduced glutathione and various enzymes catalase (CAT), glutathione reductase (GR), glutathione S- transferase (GST) and süperoxide dismutase (SOD), which are known for their detoxification capabilities in microorganisms was tested in a time depending manner for any change in their level and activity. Also, the effect of various pesticides was tested on this system. In the control groups of F. trogii grown in shaking and non-shaking conditions, the activity of CAT and GR in early stage of growth was found to be higher than the late stages and GST, SOD activities and GSH level were seen to be increased. However, while the activity of CAT was found to be increased at the late phase of growth, under shaking incubation conditions, the non-shaking incubation caused a reverse effect, mainly decreasing the activity of CAT. Under both conditions, GR, GST and SOD activities and GSH amount were reduced by the addition of the insecticide. While the CAT activity was increased at the begining of the growth, it was decreased at the 4th and 5th day of growth under shaking conditions in control group of A. niger. Also the activities of GST and GR were decreased in time but the low level of GSH at the begining of growth was seen to be increased at the 5th day of incubation. No change was observed in SOD activity, except at the late phases of growth where it was seen to be increased. The treatment of A. niger cultures with insectiside caused a decrease in GR activity and in increase in the activity of GST, CAT and SOD at the 2nd and 5lh, 4lh and 5th, 1st and 2nd days, respectively. The effect of paraquat, a herbicide, was tested on activity those enzymes and GSH levels of F. trogii. While the CAT activity and the level of GSH of F. trogii at the beginning of incubation was found to be decreased with 0.1 and 1.0 mM concentrations of paraquat, GR, SOD and GST activities were found to be increased. It was found that the CAT activity was decreased, but GR activity was increased with the application of 0.1 mM paraquat to the cultures at late growth phase. But 1.0 and 10.0 mM concentration caused a decrease in GR activity. The activity of GST was increased with the addition of herbicide at the 6th day. The level of GSH was decreased and SOD activity was increase with all three concentrations of herbicide. In general the treatment of F. trogii with paraquat caused an increased in SOD activity but a decrease in CAT activity. The accumulation of H202 after paraquat treatment may cause undesirable effect in other organisms. So, paraquat treatments should be carried out with great caution. Also, in this study, the activity of enzymes responsible for detoxification and the level of GSH, in several bacteria, were tested. There was fluctations in the activity of CAT and GR in E. coli, P. putida, S. aureus throughout the incubation time. The activity of GST and SOD and the level of GSH was seen to be decreased in time J7. trogii and those bacteria were seen to be differently behave in this respect. VI In a part of this study, we have also tested the effect of insectiside, guthion, onp. aeruginosa by the treatment of cultures both at the begining of incubation and at the late growth phases. The effect of insecticide on bacteria was studied by the addition of 5 and 20 ppm guthion into the growth media of P. aeruginosa. These concentrations caused a repression in CAT activity but an induction in GR activity. Also, while SOD activity was repressed at the 4th hour of incubation, an induction of it was observed at the later times of incubation with the application of same amount of insecticide. While the GST activity decreased the level of GSH propontionally increased. At the 8th hour of incubation, 5 and 20 ppm insecticide caused an increase at the 30 min after its addition, but a decrease was observed at the 0, 60 and 120 min of addition. While the activity of GR was increased at 30 minutes of incubations at 0 and 120 minutes of incubation. Its activity was seen to be decreased. The addition of 5 ppm caused a decreased in GST activity at 0 and 30 min, an increase in 20 ppm its activity at 60 and 120 min of incubation. Also, while the GSH level decreased the activity of SOD was observed to be increased. It was concluded that there were fluctations and differences in those systems of detoxification among microorganisms used. The treatment of those microorganisms with pesticides also caused similar changes depending mainly on the growth phase. This is a study in which the detoxification systems of microorganisms is so widely studied for the first time and also an attempt was made to figure out the changes of those agents throughout the incubation period. vıı Key Words: Süperoxide dismutase, Catalase, Glutathione reductase, Glutathione S- transferase, Glutathione, Fungus, Bacterium, Pesticide.
Açıklama
Anahtar Kelimeler
Biyoloji, Biology
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Asma, D. (1998). Bazı fungus ve bakterilerde detoksifikasyon sistemi ve sistem üzerine pestisitlerin etkisinin araştırılması. Yayımlanmış Doktora lisans tezi, İnönü Üniversitesi, Malatya.
