Abstract
The conventional method for the detection of Xanthomonas axonopodis has been
based on biochemical tests. A rapid and sensitive method for identification and
detection of Xanthomonas axonopodis is required for management of citrus
canker. PCR-based diagnostic test is appropriate for monitoring pathogen in a
very short time compared to laborious, non-specific and expensive protocols.
ELISA, Nested PCR has been used for many years in different countries. Citrus
orchards in Sargodha region were surveyed and leaf samples showing typical
symptoms of citrus canker were collected. Infected section of leaf was taken for
isolation of bacteria. Lesions were cut into parts and streak bacteria by the help of
inoculation loop grown in nutrient medium. Pure bacterial culture of Xanthomonas
axonopodis were used for detection by standard PCR. Xanthomonas axonopodis
was diagnosed by amplification of 16S rDNA. A fragment of ~1.4 kb was amplified
and cloned for sequencing Out of three markers used (K1F-CIT/ K1R-CIT, K2FCIT/ K2R-CIT, K3F-CUT/ K3R-CIY) K3F-CUT/ K3R-CIY gave best results
repeatedly. So this primer pair can be used for identification/diagnosis of
Xanthomonas axonopodis. Bacterial culture used as template in PCR and colony
PCR gave better results as compared with extracted DNA from infected leaf.
JAVERIA NEHAL, ARIF MUHAMMAD KHAN, AZHAR ABBAS KHAN, MUHAMMAD MUBIN, HAFIZ MUHAMMAD TAHIR, JAVED IQBAL. (2017) Development and Optimization of Molecular Technique for Diagnosis of Citrus Canker in Citrus Cultivars, Biologia – Journal of Biological Society of Pakistan, Volume 63 (II), Issue (II).
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