1Plant Protection Department, Kerman Agricultural and Natural Resources Research and Education Centre, Kerman, Iran
2Plant Molecular Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
3Advanced Centre for Plant Virology, Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India
*Corresponding author e-mail: Mehdi Azadvar (mehdiazadvar@gmail.com)
Online published on 23 May, 2023.
Witches’ broom (WB) and “huanglongbing” (HLB) associated to phloem limited bacteria ‘Candidatus Phytoplasma aurantifolia’ and ‘Ca. Liberibacter asiaticus’, respectively are the most serious and destructive citrus diseases in the south of Iran. Although it is possible to detect each of these pathogens by PCR separately, this is expensive and time-consuming. In this study a duplex PCR detection system using P4/P7 and fp400/rp400 primer pair sets for simultaneous detection of both pathogens was developed. Two 400 and 500 bp amplicons were simultaneously amplified and no bands were detected from healthy plants. The consistent results of the developed duplex PCR were compared with simplex PCR for detection of each pathogen and confirmed with sequencing analysis. This rapid, sensitive, specific and robust duplex PCR assay should be useful for detection of WB and HLB especially for quarantine purposes, disease surveys, certification and plant improvement programs for obtaining pathogenfree plant materials..
‘Candidatus Phytoplasma aurantifolia’, ‘Candidatus Liberibacter asiaticus’, Polymerase chain reaction
