Identification and quantification of Aphelenchoides besseyi from rice using qPCR

Celik E. S., DEVRAN Z.

EUROPEAN JOURNAL OF PLANT PATHOLOGY, cilt.154, sa.3, ss.691-703, 2019 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 154 Sayı: 3
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1007/s10658-019-01692-4
  • Dergi Adı: EUROPEAN JOURNAL OF PLANT PATHOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.691-703
  • Anahtar Kelimeler: Detection, qPCR, Quantification, White tip nematode, REAL-TIME PCR, WHITE TIP NEMATODE, GLOBODERA-ROSTOCHIENSIS, PINEWOOD NEMATODE, PLANT-PARASITISM, SYBR GREEN, TAQMAN, ASSAY, DIFFERENTIATION
  • Akdeniz Üniversitesi Adresli: Evet

Özet

Rice is an essential food source for humans. Rice white tip nematode, Aphelenchoides besseyi, is the most important and devastating plant-parasitic nematode of rice. In order to prevent the spread of A. besseyi, a rapid and accurate diagnostic protocol is required for seed certification and quarantine purposes. Here, we developed a qPCR assay based on small subunit ribosomal DNA (SSU rDNA) of A. besseyi and validated it successfully in white tip nematode populations collected from rice-growing fields. Our findings indicate that a dilution series of plasmid DNA of A. besseyi showed a high correlation (R-2=0.99, efficiency 95%) with Ct values. The different stages of white tip nematode, i.e. egg, juvenile, male and female, were successfully identified using qPCR assay; the single juvenile nematode was also detected without separate DNA isolation. In addition, qPCR assay without the need for a separate nematode extraction step directly identified A. besseyi from infected brown rice, hull and paddy. Since the novel qPCR assay is specific and sensitive for detection of A. besseyi, it could be used for detection and quantification of rice white tip nematode.