In the central and south eastern provinces of Iran, alfalfa plants showing phytoplasma diseases-like symptoms categorized into three major types, namely witches'-brooms, little leaf and yellowing, were observed during 2008 and 2009. Phytoplasmas were detected by PCR assays using phytoplasma universal primer pair P1/P7 followed by nested PCR using primer pair R16F2n/R16R2. PCR results confirmed the phytoplasma infection in symptomatic alfalfa plants. Restriction fragment length polymorphism (RFLP) and sequence analysis of partial 16S rRNA gene from the Iranian alfalfa phytoplasmas indicated that phytoplasmas of the 16SrII and 16SrXII groups were associated with alfalfa witches'-broom and alfalfa yellows, respectively, while phytoplasmas of the 16SrII-D subgroup are associated with alfalfa little leaf disease. This research is the first study to report stolbur phytoplasmas as causal agents of alfalfa yellows and 16SrII-D phytoplasmas as causal agents of alfalfa little leaf diseases in Iran.

White clover plants showing little leaf and leaf reddening symptoms were observed in Isfahan Province in central Iran. Restriction fragment length polymorphism analyses of nested PCR‐amplified fragments from Iranian clover little leaf phytoplasma isolates and representative phytoplasmas from other phytoplasma groups using AluI, CfoI, KpnI and RsaI restriction enzymes indicated that the clover phytoplasma isolates are related to the peanut WB group. Sequence analyses of partial 16S rRNA fragments showed that Iranian clover little leaf phytoplasma has 99% similarity with soybean witches'‐broom phytoplasma, a member of the peanut WB (16SrII) phytoplasma group. This is the first report of clover infection with a phytoplasma related to the 16SrII group.