PJB-2016-248
DETECTION AND MOLECULAR CHARACTERIZATION OF CANDIDATUS LIBERIBACTER SPP. CAUSING HUANGLONGBING (HLB) IN INDIGENOUS CITRUS CULTIVARS IN PAKISTAN
AMBREEN ZAFARULLAH, SHAGUFTA NAZ* AND FAIZA SALEEM
Abstract
Citrus greening or huanglongbing (HLB) is one of major devastating citrus diseases all over the world. This disease is caused by fastidious α-proteobacterium, Candidatus liberibacter spp. and is transmitted by grafting as well as psyllids Diaphorina citri or Trioza erytreae. The objective of this study was to identify and characterize the huanglongbing (HLB) infectious pathogen in commercial (Kinnow and sweet oranges) varieties by using molecular markers such as 16S rRNA, 16S/23S rRNA and outer membrane protein fragments from symptomatic leaves of assorted citrus varieties. DNA extracted from forty different citrus (including mandarin and sweet oranges) varieties having HLB-symptomatic plants from different orchards of Pakistan. Gene-specific primers for 16SrDNA, 16S/23S rDNA and outer membrane protein (OMP) gene regions were used for identification of Ca. liberibacter spp. An amplified fragment of 1174 bp from 16SrDNA, 900 bp of 16S/23S rRNA and 600 bp was observed for OMP gene fragments of Asian isolates. The resulted fragments were TA cloned and sequenced from both strands. The infectious bacterium was identified as Candidatus liberibacter asiaticus and was found in 17 samples (42%). The seasonal variation on prevalence of Candidatus liberibacter asiaticus in citrus varieties was well observed. It declined during spring season due to unfavourable temperature and humidity for Candidatus liberibacter asiaticus because disease symptoms showed mostly at low humidity and warm temperature (up to 35°C). Key words: Huanglongbing, Candidatus liberibacter asiaticus, Outer membrane protein, Diaphorina citri, α-proteobacterium.
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