Hatay ilinde zeytin dal kanseri (Pseudomonas savastanoi pv. savastanoi) hastalığının tespiti ve biyolojik mücadele olanaklarının araştırılması
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Dosyalar
Tarih
2020
Yazarlar
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Yayıncı
Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışma ile Hatay ili zeytin üretim alanlarında zeytin dal kanseri hastalığının yaygınlık ve bulunma oranları belirlenmiş ve sağlıklı zeytin ağaçlarının kök ve sürgünlerinden izole edilen epifit ve endofit bakteriler ile Pseudomonas savastanoi pv. savastanoi'nin biyolojik mücadele olanakları araştırılmıştır. Hastalığın en yüksek yaygınlık oranı %90 ile Samandağ, İskenderun ve Arsuz ilçelerinde belirlenmiş olup, Hassa, Kırıkhan, Reyhanlı ve Kumlu ilçelerinde ise hastalık saptanmamıştır. Hastalığın enfekteli bahçelerdeki bulunma oranları ise %4.5-75 arasında değişiklik göstermiştir. Taze urlardan izole edilen hastalık etmeninin tanısı Gram boyama, LOPAT testleri, MALDI-TOF ve moleküler yöntemler ile yapılmıştır. Yapılan testler sonucu toplam 14 izolat Pseudomonas savastanoi pv. savastanoi olarak tanılanmıştır. Biyolojik mücadele çalışmalarında 218 epifit ve 144 endofit olmak üzere toplam 362 bakteri izole edilmiş ve MALDITOF-MS ile tanılanmıştır. Antagonistik etkinin (A-indeks) belirlendiği ikili kültür denemelerinde 81 epifit ve endofit bakteri izolatı patojen gelişimini farklı oranlarda engellemiştir. En yüksek engelleme oranı 5.24 A-indeks değeri ile Bacillus megaterium HZEP7 izolatında belirlenirken bunu sırası ile Bacillus subtilis HZEN1 (4.85) ve Pseudomonas koreensis HZEN27 (4.83) izolatları izlemiştir. İn vitro etki mekanizmalarının belirlenmesi çalışmalarında epifit ve endofit bakterilerin indol asetik asit (IAA) üretimi, siderofor üretimi, fosfor çözme potansiyellerinin belirlenmesi, proteaz üretimi ve amonyak üretimi gibi mekanizmaları incelenmiştir. İn vivo testlerde kullanılacak izolatlar yüksek antagonistik etki, düşük IAA üretimi ve diğer etki mekanizmalarında göstermiş oldukları etkiye göre belirlenmiş olup, Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 ve Bacillus pumilus HZEP29 izolatları havuç dilimleri ve fidan inokulasyonu ile biyokontrol etkinin belirlenmesi çalışmalarına seçilmiştir. Havuç dilimleri inokulasyonu ile biyokontrol etkinin belirlenmesi testlerinde, en yüksek engelleme % 98,78 ile Bacillus subtilis HZEN1 izolatında belirlenirken bunu sırası ile Bacillus megaterium HZEP7 (%94.49), Pseudomonas koreensis HZEN27 (%92.09) ve Bacillus pumilus HZEP29 (%90.26) izolatları izlemiştir. Fidan inokulasyonu testlerinde havuç dilimlerine benzer şekilde tüm bakteri izolatları ur oluşumunu engellemiş olup en yüksek engelleme %78.72 ile Bacillus subtilis HZEN1 izolatında belirlenirken bunu sırası ile Bacillus megaterium HZEP7 (60.87), Pseudomonas koreensis HZEN27 (46.09) ve Bacillus pumilus HZEP29 (44.87) izolatları izlemiştir. Bakır sülfat uygulamasında ise ur oluşumu %48.23 oranında engellenmiştir. Elde edilen sonuçlar Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 ve Bacillus pumilus HZEP29 izolatlarının zeytin dal kanseri hastalığı ile mücadelede biyokontrol etmeni olarak kullanılabileceğini göstermiştir.
The aim of this study was to determine the prevalence and incidence of olive knot disease in Hatay province and the biological control possibilities of Pseudomonas savastanoi pv. savastanoi by using epiphyte and endophyte bacteria isolated from the roots and shoots of healthy olive trees. The highest prevalence rate of the disease was determined in Samandağ, İskenderun and Arsuz districts with 90 % and no disease incidence was detected in Hassa, Kırıkhan, Reyhanlı and Kumlu districts. The incidence of the disease in infected gardens varied between 4.5-75%. The causal disease agent was identified by Gram stain, LOPAT, MALDI-TOF and molecular techniques. All strains were determined as gram negative, levan negative, oxidase negative, pectolytic activity negative, arginine dehydrolase negative, but tobacco hypersensitivity reaction was positive. According to the test results, a total of 14 isolates were identified as Pseudomonas savastanoi pv. savastanoi. In biological control studies, a total of 362 bacteria (218 epiphytes and 144 endophytes) were isolated and diagnosed by using MALDITOF-MS. In dual culture tests, 81 epiphyte and endophyte bacterial isolates inhibited pathogen development at different rates. The highest inhibition rate (A-indeks) was determined in Bacillus megaterium HZEP7 isolate with 5.24 A-index value, followed by Bacillus subtilis HZEN1 (4.85) and Pseudomonas koreensis HZEN27 (4.83), respectively. Mechanisms of epiphyte and endophyte bacteria such as indole acetic acid (IAA) production, siderophore production, determination of phosphorus dissolving potentials, protease production and ammonia production were investigated by in vitro tests. The isolates to be used in in vivo tests have been determined according to high antagonistic effect, low IAA production and other mechanisms, and Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 and Bacillus pumilus HZEP29 isolates have been selected for determination of biocontrol effect by carrot slices and seedlings inoculation. In the tests for determination of the biocontrol effect by carrot slices inoculation, the highest inhibition was detected in Bacillus subtilis HZEN1 isolate with 98.78%, followed by Bacillus megaterium HZEP7 (94.49%), Pseudomonas koreensis HZEN27 (92.09%) and Bacillus pumilus HZEP29 (90.26%) isolates, respectively. Similar to carrot slices in seedling inoculation tests, all bacterial isolates inhibited gall formation. The highest inhibition was determined in Bacillus subtilis HZEN1 isolate with 78.72%, followed by Bacillus megaterium HZEP7 (60.87%), Pseudomonas koreensis HZEN27 (46.09%) and Bacillus pumilus HZEP29 (44.87%) isolates, respectively. In addition, the gall formation was inhibited by 48.23% in copper sulphate application. The obtained results showed that Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 and Bacillus pumilus HZEP29 isolates could be used against olive knot disease as a biocontrol agent.
The aim of this study was to determine the prevalence and incidence of olive knot disease in Hatay province and the biological control possibilities of Pseudomonas savastanoi pv. savastanoi by using epiphyte and endophyte bacteria isolated from the roots and shoots of healthy olive trees. The highest prevalence rate of the disease was determined in Samandağ, İskenderun and Arsuz districts with 90 % and no disease incidence was detected in Hassa, Kırıkhan, Reyhanlı and Kumlu districts. The incidence of the disease in infected gardens varied between 4.5-75%. The causal disease agent was identified by Gram stain, LOPAT, MALDI-TOF and molecular techniques. All strains were determined as gram negative, levan negative, oxidase negative, pectolytic activity negative, arginine dehydrolase negative, but tobacco hypersensitivity reaction was positive. According to the test results, a total of 14 isolates were identified as Pseudomonas savastanoi pv. savastanoi. In biological control studies, a total of 362 bacteria (218 epiphytes and 144 endophytes) were isolated and diagnosed by using MALDITOF-MS. In dual culture tests, 81 epiphyte and endophyte bacterial isolates inhibited pathogen development at different rates. The highest inhibition rate (A-indeks) was determined in Bacillus megaterium HZEP7 isolate with 5.24 A-index value, followed by Bacillus subtilis HZEN1 (4.85) and Pseudomonas koreensis HZEN27 (4.83), respectively. Mechanisms of epiphyte and endophyte bacteria such as indole acetic acid (IAA) production, siderophore production, determination of phosphorus dissolving potentials, protease production and ammonia production were investigated by in vitro tests. The isolates to be used in in vivo tests have been determined according to high antagonistic effect, low IAA production and other mechanisms, and Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 and Bacillus pumilus HZEP29 isolates have been selected for determination of biocontrol effect by carrot slices and seedlings inoculation. In the tests for determination of the biocontrol effect by carrot slices inoculation, the highest inhibition was detected in Bacillus subtilis HZEN1 isolate with 98.78%, followed by Bacillus megaterium HZEP7 (94.49%), Pseudomonas koreensis HZEN27 (92.09%) and Bacillus pumilus HZEP29 (90.26%) isolates, respectively. Similar to carrot slices in seedling inoculation tests, all bacterial isolates inhibited gall formation. The highest inhibition was determined in Bacillus subtilis HZEN1 isolate with 78.72%, followed by Bacillus megaterium HZEP7 (60.87%), Pseudomonas koreensis HZEN27 (46.09%) and Bacillus pumilus HZEP29 (44.87%) isolates, respectively. In addition, the gall formation was inhibited by 48.23% in copper sulphate application. The obtained results showed that Bacillus subtilis HZEN1, Bacillus megaterium HZEP7, Pseudomonas koreensis HZEN27 and Bacillus pumilus HZEP29 isolates could be used against olive knot disease as a biocontrol agent.
Açıklama
Anahtar Kelimeler
Ziraat, Agriculture
