Isolation and Identification of Bacterial Soft Rot Disease Agent Pectobacterium Isolates on Potato Tubers Delivered for Consumption in Yozgat and Kırşehir Provinces

Abstract

Pectobacterium spp. are ubiquitous bacterial plant pathogens with a wide host range which cause soft rot disease on the invaded plant tissue. This study aimed to isolate the bacterial soft rot disease agents belonging to Pectobacterium spp. on potato tubers delivered for consumption in the local bazaar, grocery and markets in the Center district of Yozgat province and Kaman district of Kırşehir province of Turkey between 2018 and 2019. Local bazaar, market and grocery stores were surveyed and 168 potato tubers were collected. The humidity cell test was performed to investigate the presence of pectinolytic bacteria in asymptomatic and soft rotted potato tubers. Bacterial isolation from 54 different soft rotted tuber slices was performed on Crystal Violet Pectate (CVP) medium. A total of 42 bacterial isolates, that formed pits on CVP medium, were obtained following isolation. Among these, 27 isolates were observed to have shiny, transparent, irregular, crater-shaped colony morphology on Nutrient Agar (NA) plates. All isolates were gram and oxidase negative, catalase-positive, facultative anaerobe, non-fluorescent morphological characteristics and caused soft rot on potato tuber slices. Following PCR analysis, 19 of 54 symptomatic tuber samples were determined as infected by P. carotovorum subsp. carotovorum (9 isolates), P. atrosepticum (6 isolates) and Pectobacterium spp. (4 isolates). In 8 of 114 asymptomatic tuber samples, P. carotovorum subsp. carotovorum (5 isolates), P. atrosepticum (2 isolates) and P. carotovorum subsp. brasiliense (1 isolate) were determined as responsible agents of latent infections. The presence of Pectobacterium isolates in the stored potato tubers brought for consumption suggests the pathogen can be transferred to different regions. Potato tubers offered for sale for consumption are thought to be an important inoculum source for the transfer of Pectobacterium isolates to disease-free areas.

Keywords

• Pectobacterium spp.
• Soft Rot
• Latently Infected Tuber
• Potato
• PCR

Project Number

6602c-ZF/18-237

Yozgat ve Kırşehir İllerinde Tüketime Sunulmuş Patates Yumrularında Bakteriyel Yumuşak Çürüklük Hastalığı Etmeni Pectobacterium İzolatlarının İzolasyonu ve Tanılanması

Abstract

Pectobacterium spp. geniş bir konukçu yelpazesine sahip, her ortamda bulunan bakteriyel bitki patojenlerden biri olup, istila ettiği bitki dokusunda Bakteriyel Yumuşak Çürüklük olarak bilinen hastalığa neden olur. Bu çalışmada 2018-2019 yılları arasında Yozgat Merkez ve Kırşehir Kaman ilçesindeki yerel pazar, manav ve zincir marketlerde satışa sunulan patates yumrularında bakteriyel yumuşak çürüklük hastalığına neden olan Pectobacterium spp. izolasyonu, biyokimyasal, patojenisite ve moleküler yöntemlerle tanılanması amaçlanmıştır. Survey yapılan market ve manavlardan 168 yumru örneği toplanmıştır. Tipik yumuşak çürüklük belirtisi gösteren ve göstermeyen şüpheli yumrularda pektolitik izolatların varlığının araştırılması amacıyla nemli hücre testi yapılmıştır. Çürüme belirtileri gösteren 54 farklı yumru kesitlerinden pektolitik izolatların izolasyonu Kristal Violet Pektat (CVP) besiyeri üzerinde yapılmıştır. CVP besi yerinde çukur oluşturan pektolitik koloniler NA besi yerine saflaştırılması ile toplam 42 adet bakteri izolatı elde edilmiştir. Elde edilenler bakteri izolatları arasında 27 farklı izolatın Nutrient Agar (NA) besi yerinde şeffaf, parlak, düzensiz, krater formda koloni oluşturduğu belirlenmiştir İzolatların tamamı gram ve oksidaz negatif, katalaz pozitif, fakültatif anaerob, floresen olmayan koloni morfolojisine sahip özelliklerde sergilemiş ve patates dilimi üzerinde yumuşak çürüklüğe neden olmuştur. Elde edilen izolatlar ile yapılan PCR çalışmasında, 54 simptomlu yumru örneğinin 19 adedinde P. carotovorum subsp. carotovorum (9 izolat), P. atrosepticum (6 izolat) ve Pectobacterium spp. (4 izolat) enfeksiyona neden olan etmenler olarak tanılanmıştır. Tipik hastalık belirtisi göstermeyen 114 yumru örneğinin 8 adedinde P. carotovorum subsp. carotovorum (5 izolat), P. atrosepticum (2 izolat) ve P. carotovorum subsp. brasiliense (1 izolat) etmenleri latent enfeksiyon sorumlusu olarak belirlenmiştir. Bu çalışma ile tüketime yönelik olarak satışa sunulmuş patates yumrularında Pectobacterium izolatlarının varlığı belirlenerek patojenin farklı bölgelere transfer edilebileceği belirlenmiştir. Tüketim için satışa sunulan patates yumrularının hastalıktan ari bölgelere Pectobacterium izolatlarının nakledilmesinde önemli bir inokulum kaynağı olduğu düşünülmektedir.

Keywords

• Pectobacterium spp.
• Yumuşak Çürüklük
• Latent Enfekteli Yumru
• Patates
• PCR

Supporting Institution

Yozgat Bozok Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi

Project Number

6602c-ZF/18-237

Thanks

Bu çalışma, Yozgat Bozok Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi 6602c-ZF/18-237 nolu proje bütçesinden desteklenmiştir.

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