Molecular characterization, vector identification and partial host range determination of phytoplasmas associated with faba bean phyllody in Iran

Abstract

During surveys of phytoplasma diseases, faba bean phyllody (FBP) was observed in several locations in Fars and Bushehr provinces (southern Iran). Samples of affected plants from Borazjan (Bushehr province) and Fasa (Fars province) were used to transmit the phyllody agent to faba bean, mung bean, pea, alfalfa and periwinkle by grafting, dodder and/or vector insect. Orosius albicinctus (Distant) was identified as the vector of Borazjan (BFBP) and Fasa (FFBP) faba bean phyllody. Naturally affected faba bean plants and all inoculated plants were positive for phytoplasma by direct PCR using the P1/P7 primer pair and nested PCR using P1/P7 and R16F2n/R16R2 primer pairs. Sequencing of PCR products identified the associated phytoplasmas as members of 16SrII phytoplasma group. Phylogenetic analysis using full length 16S rRNA gene sequences also confirmed similarity of the BFBP and FFBP phytoplasmas to the 16SrII group phytoplasmas. In this analysis the FFBP phytoplasma was grouped with ’Candidatus Phytoplasma australasia’, representative of 16SrII-D subgroup, while the BFBP phytoplasma formed a discrete group close to the 16SrII-C subgroup. Restriction fragment length polymorphism (RFLP) confirmed that BFBP and FFBP phytoplasmas belong to 16SrII group. Virtual RFLP confirmed that as members of peanut witches’ broom (16SrII) phytoplasma group, BFBP and FFBP phytoplasmas belonged to 16SrII-C and16SrII-D subgroups, respectively. Phytoplasmas associated with BFBP and FFBP were shown to be serologically related to the Fars alfalfa witches’broom phytoplasma, a member of 16SrII-C subgroup. It seems that witches’broom affected alfalfa fields are natural reservoirs of the FBP phytoplasma in southern Iran.