Molecular detection of Candidatus liberibacter asiaticus associated with Citrus greening (Huanglongbing) of mandarin by DNA template preparation
Keywords:
PCR, HLB, CitrusAbstract
The polymerase Chain Reaction (PCR) diagnosis is more reliable and sensitive diagnostic tool for detecting greening bacterium than other conventional approaches like Electron microscopy, DNA-DNA hybridization and immunofluorescence (IF) for detection of citrus greening. During experiments, it was observed that sodium sulphite method of DNA isolation provided higher yield and better quality DNA than other methods. Primer C (450 bp) was more efficient in amplifying the DNA of greening bacterium even at a very low concentration of 0.1 pg. To confirm the reliability of PCR, the greening bacterium was also detected in graft-inoculated plants, which showed typical greening bacterium was also detected in graft-inoculated plants, which showed typical greening symptoms. Results showed amplification of 450 bp in PCR suggesting sampling in March is more suitable for PCR detection of greening bacterium.References
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