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Detection of candidatus liberibacter asiaticus associated with citrus greening (huanglongbing) of mandarin by template preparation |
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ABSTRACTThe polymerase ChainReaction (PCR) diagnosis is more reliable and sensitive diagnostic tool forgreening bacterium than other conventional approaches like Electron microscopy,DNA-DNA hybridization and immunofluorescence (IF) for detection of citrusgreening. During experiment, it was observed that sodium sulphite method of DNAisolation provided higher yield and better quality DNA than other methods.Primer C (450 bp) was more efficient in amplifying the DNA of greeningbacterium even at a very low concentration of 0.1 pg. To confirm thereliability of PCR, the greening bacterium was also detected ingraft-inoculated plants, which showed typical greening bacterium was alsodetected in graft-inoculated plants, which showed typical greening symptoms.Results showed amplification of 450 bp in PCR suggesting sampling in March ismore suitable for PCR detection of greening bacterium. |