Detection and molecular characterization of tomato big bud disease in Qazvin province

Some samples were collected from tomato fields in Qazvin from tomato plants with big bud symptoms such as plant droop and purplish vein under the leaf, enlarged and sac-like pistils and malformed buds. DNA was extracted from the veins and vascular tissues of the plant with CTAB-based methods. In symptomatic plants, DNA fragments of ۱۸۰۰ and ۱۲۰۰bp were amplified by PCR using P۱/P۷, R۱۶F۲n/R۱۶R۲ primers. Restriction fragment length polymorphism (RFLP) analysis of nested R۱۶F۲n/R۱۶R۲ primed PCR product (۱۲۰۰bp) showed that the tomato big bud phytoplasma from Qazvin (TOM-Qazvin) is a member of clover proliferation (۱۶SrVI). Phylogenetic analysis of ۱۶SrRNA and putative restriction site analysis of the R۱۶F۲n/R۱۶R۲ primed sequence classified TOM phytoplasma in clover proliferation (۱۶SrVI) group and belonged to subgroup ۱۶SrVI-A. Virtual RFLP by using ۱۲۰۰bp sequencing of ۱۶SRNA and ۱۷ restriction enzymes confirmed that TOM-Qazvin belonged to the subgroup ۱۶SrVI-A and۱۶SrVI group. To our knowledge, this is the first report of tomato big bud disease in Qazvin province.

کلیدواژه ها:

big bud ، iphyclassifier ، RFLP ، tomato ، phytoplasma