A real-time PCR assay using locked nucleic acid probe for detection of Acidovorax citrulli

ÖZTÜRK N., BASIM H.

JOURNAL OF PLANT DISEASES AND PROTECTION, cilt.129, sa.2, ss.395-409, 2022 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 129 Sayı: 2
  • Basım Tarihi: 2022
  • Doi Numarası: 10.1007/s41348-021-00559-3
  • Dergi Adı: JOURNAL OF PLANT DISEASES AND PROTECTION
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, BIOSIS, CAB Abstracts, Geobase
  • Sayfa Sayıları: ss.395-409
  • Anahtar Kelimeler: Acidovorax citrulli, Watermelon Bacterial Fruit Blotch, Quantitative real-time PCR, Detection, AVENAE SUBSP CITRULLI, BACTERIAL FRUIT BLOTCH, POLYMERASE-CHAIN-REACTION, CAUSAL AGENT, QUANTITATIVE DETECTION, SENSITIVE DETECTION, PV. MANIHOTIS, WATERMELON, MELON, STRAINS
  • Akdeniz Üniversitesi Adresli: Evet

Özet

Watermelon Bacterial Fruit Blotch disease caused by Acidovorax citrulli is an economically important disease to the watermelon plant. The primary source of infection for A. citrulli is the internally infected seeds of watermelon. In this study, a real-time PCR assay was developed for the detection of A. citrulli from watermelon leaves and seed samples. Primer and locked nucleic acid (LNA) probe sets were designed using the hrpB2 sequence and the ITS (internally transcribed spacer). These primer-probe sets amplified the 88 bp. Detections of A. citrulli strains were performed by a real-time PCR method using the designed primers and LNA probe sets in 15-26 min. The sensitivity limit for bacterial cells was 2 bacterial cells, and the sensitivity limit at DNA level was 12 fg. The efficiency of the real-time PCR for A. citrulli was highly reliable with rates of 98.35% and 99.18% for hrpB2 and ITS, respectively. The real-time PCR assay using the LNA probe is the first study to demonstrate the feasibility of detecting different strains of A. citrulli from the diseased watermelon leaves and the infected seeds.