تجزیه توالی چند لوکوسی جدایه های Candidatus liberibacter asiaticus در ایران

Citrus Huanglongbing (HLB), Huanglongbing (HLB), is one of the most destructive citrus diseases in the world. To amplify partial CLas housekeeping genes, six primer pairs based on the Psy62 isolate were designed using Vector NTI 9.2 software and Primer3 4.1.0 and synthesized by Metabion (Metabion©, Germany). The rpoB, dnaG, rplA, mutS, fumC and nusG genes were amplified in 27 isolates using polymerase chain reaction (PCR). The amplified fragments of different genes of different CLas isolates were sent directly to the genetic codon company (Codon Genetic Group©, Iran) for sequencing. Sequences were edited using Geneious Prime 2020.1.2 software and deposited in the national center for biotechnology information (NCBI) GenBank. Phylogenetic relationship of the isolates based on the concatenated partial gene sequences of dnaG, fumC, mutS, nusG, rplA and rpoB (4305 bp) was investigated. Nucleotide alignment of the sequences was performed using the ClustalW software. In order to determine the phylogenetic position of the isolates, maximum similarity, maxiumum parsimony and neighbour-joining algorithms were used. The neighbour-joining model was selected by jModeltest based on the lowest BIC, the TVMef model was selected by estimating the gamma distribution, and the maximum similarity dendrogram was plotted by RAxML 8 with 1000 Bootstrap replications. The neighbour-joining method was performed using the p-distance model with 1000 Bootstrap with MEGAX software. The maxiumum parsimony analysis was run with heuristic searches with TBR branch swapping, and random addition of taxa and 1000 bootstraps replicates using PAUP* 4.0a. The mixed dendrogram was plotted by adding the reproducibility of the Neighbour-joining methods and the maxiumum parsimony to the maximum similarity tree. In the cumulative lineage tree of housekeeping genes, Iranian CLas isolates and CLas isolates in the gene bank were grouped together in separate groups. Also, some typical genera of the Rhizobiaceae family of the order Hyphomicrobiales include Agrobacterium fabrum str. C58 and Rhizobium etli CFN42 and Bartonella bacilliformis KC583 from Bartonellaceae family and Hyphomicrobiales were bunched in the separate groups. The reference species Escherichia coli str Sakai from Gammaproteobacteria group was also placed in a separate group.