Affordable Access

Development Of A New Probe For Specific And Sensitive Detection Of 'Candidatus Phytoplasma Mali' In Inoculated Apple Trees

Publication Date
  • Life Sciences :: Phytobiology (Plant Sciences
  • Forestry
  • Mycology...) [F12]
  • Sciences Du Vivant :: Biologie Végétale (Sciences Végétales
  • Sylviculture
  • Mycologie...) [F12]
  • Design


aab_171 251..258 RESEARCH ARTICLE Development of a new probe for specific and sensitive detection of ‘Candidatus Phytoplasma mali’ in inoculated apple trees M. Aldaghi1,2,*, S. Massart1,*, S. Roussel1 & M.H. Jijakli1 1 Plant Pathology Unit, Gembloux Agricultural University, Gembloux, Belgium 2 Plant Diseases Research Department, Plant Pests & Diseases Research Institute, Tehran, Iran *These authors equally contributed to this paper. Keywords ‘Candidatus Phytoplasma mali’; detection; real-time PCR; sensitivity. Correspondence M.H. Jijakli, Plant Pathology Unit, Gembloux Agricultural University, Passage des de´porte´s 2, 5030 Gembloux, Belgium. Email: [email protected] Received: 7 December 2006; revised version accepted: 11 June 2007. doi:10.1111/j.1744-7348.2007.00171.x Abstract A new TaqMan minor groove binding (MGB) probe and new PCR conditions were designed for quick, specific and sensitive detection of ‘Candidatus Phyto- plasma mali’. The new probe can distinguish a single mismatch between ‘Ca. P. mali’ and ‘Candidatus Phytoplasma prunorum’, this constituting an improve- ment over a previously published method. In our study, the relative position of the mismatch in the MGB probe influenced greatly the specificity of detec- tion. Our new real-time PCR protocol was able to detect one plasmid copy in water and 100 copies in healthy plant DNA extracts. The sensitivity of this new real-time PCR method, three other real-time PCR protocols and a conventional PCR with fU5/rU3 primers was compared. Periwinkles and MM106 rootstocks were grafted with infected material and surveyed over time by symptom observation, conventional PCR and real-time PCR. Phytoplasma infection was detected by symptom observation in all periwinkles within 4 months and in 75% of the MM106 rootstocks by the end of 7 months. Conventional PCR de- tected phytoplasma infection in all periwinkles within 4 months and in all MM106 rootstocks within 7 months. Best resu

There are no comments yet on this publication. Be the first to share your thoughts.