Fasulye bitkisinden elde edilen endofit bakterilerin bitki gelişimini teşvik edici ve antagonistik özelliklerinin Pseudomonas syringae pv. phaseolicola etmenine karşı karekterizasyonu

Öz

Pseudomonas syringae pv. phaseolicola (Psp) fasulye bitkisinde Hale yanıklığı hastalığına sebep olan tohum kökenli bir bakteriyel hastalık etmenidir. Bu çalışmanın amacı, farklı bölgelerde yetişen sağlıklı fasulye bitkilerinden izole edilen endofitik bitki gelişimini teşvik eden bakterilerin (PGPB) Psp üzerine antagonistik (siderofor ve amonyak üretimi gibi) ve bitki gelişimini teşvik eden mekanizmalarının (IAA üretimi, fosfatı indirgeme özelliği gibi) in vitro koşullarda karakterizasyonudur. Araştırma sonunda elde edilen 131 PGPB arasında 71 izolat Psp etmenine karşı değişen oranlarda engelleme zonu oluşturmuş olup, bu izolatları temsilen 30 izolatın teşhisleri, antagonistik ve bitki gelişimini teşvik eden mekanizmaları araştırılmıştır. Morfolojik, fizyolojik, biyokimyasal ve MALDI-TOF analizler, elde edilen izolatların 10 cins 24 farklı türe ait olduğunu göstermiştir. Bu izolatlar arasında 10 izolat Bacillus spp., 6 izolat Pseudomonas spp., 4 izolat Rhizobium radiobacter, 2 izolat Arthrobacter spp., 2 izolat Achromobacter spanius, 2 izolat Serratia liquefaciens, 1’er izolat Acinetobacter calcoaceticus, Exiguobacterium sp., Microbacterium hydrocarbonoxydans ve Ochrobactrum anthropi olarak teşhis edilmiştir. Psp’ye karşı en yüksek engelleme zonu Pseudomonas gessardii (4.85) izolatı tarafından, en düşük engelleme zonu ise Bacillus licheniformis (1.35) tarafından oluşturulmuştur. Test edilen izolatlardan 10 izolat α-amylase pozitif, 7 izolat fosfataz pozitif, 29 izolat siderofor üretmiş, 11 izolat proteaz pozitif bulunmuştur. Tüm izolatlar IAA ve amonyak üretimi açısından değişken değerlerde pozitif bulunmuştur. En yüksek siderofor üretimi P. gessardii (5.83 indeks değeri), en yüksek protease üretimi Exiguobacterium sp. (5.25 indeks değeri), en yüksek IAA üretimi P. gessardii ve O. anthropi izolatları (161.39 µg/ml), en yüksek fosfor çözme kapasitesi Acinetobacter calcoaceticus izolatı (2.63 indeks değeri) tarafından gösterilmiştir. Bu çalışma ile ilk kez fasulye bitkisinde bitki gelişimini teşvik eden ve Psp’ye antagonistik etkiye sahip endofit bakteriler elde edilmiştir.

Anahtar Kelimeler

• fasulye,hale yanıklığı,antagonist,bitki gelişimi teşvik eden bakteriler,endofit,biyolojik mücadele,Pseudomonas syringae pv. phaseolicola

Characterization of plant growth-promoting traits and antagonistic potentials of endophytic bacteria from bean plants against Pseudomonas syringae pv. phaseolicola

Abstract

Pseudomonas syringae pv. phaseolicola (Psp) is a seed-borne bacterium which causes halo blight disease in common bean. This study aimed the determination of plant growth-promoting traits (such as IAA, phosphate solubilization) and antagonistic potentials (such as siderophore and ammonia productions) of endophytic plant growth-promoting bacteria (PGPB) from healthy bean plants growing in different regions against Psp in vitro conditions. A total of 131 bacteria were primarily characterized as putative PGPB and tested for antagonist activity against Psp in dual culture tests. Seventy-one bacterial isolates demonstrated antagonistic activities against Psp isolate in varying ratios. Among these 71 isolates, 30 representative bacterial isolates from the different regions/fields were selected. On the basis of morphological, physiological, biochemical characteristics and confirmation by MALDI-TOF analyses, 30 endophytic antagonist isolates were identified as belonging to 10 genera, 24 different species. According to results obtained, 10 isolates belong to Bacillus spp., 6 isolates belong to Pseudomonas spp., 4 isolates belong to Rhizobium radiobacter, 2 isolates belong to Arthrobacter spp., 2 isolates belong to Achromobacter spanius, 2 isolates belong to Serratia liquefaciens, 1 isolate belongs to Acinetobacter calcoaceticus, Exiguobacterium sp., Microbacterium hydrocarbonoxydans, and Ochrobactrum anthropi. The largest and lowest inhibition zone was produced by endophytic bacterial isolates Pseudomonas gessardii (4.85) and Bacillus licheniformis (1.35). Among the tested antagonist bacterial isolates, 10 isolates were positive for the production of α-amylase, 7 isolates positive for phosphate solubilization, 29 isolates positive for siderophore production, 11 isolates positive for protease production. All selected bacterial isolates produced IAA and ammonia in relatively varying amounts. P. gessardii produced a relatively large amount of extracellular siderophore (5.83), Exiguobacterium sp. produced a relatively large amount of extracellular protease (5.25), P. gessardii and O. anthropi produced a relatively large amount of extracellular IAA (161.39 µg/ml) and Acinetobacter calcoaceticus produced a relatively large amount of phosphatase (2.63). This is the first study reporting bean plants harbor endophytes having plant growth promoting activities with antagonistic potential against Psp.

Keywords

• bean,halo blight,endophyte,antagonist,plant growth promoting bacteria,biological control,Pseudomonas syringae pv. phaseolicola

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