Identification of a novel phytoplasma (16Sr XIV-A subgroup) associated with little leaf and witches' broom of Cannabis sativa L. ssp sativa and C-sativa L. ssp indica in India

摘要

Symptoms of yellowing, little leaf and witches' broom on Cannabis sativa L. ssp. sativa and C. sativa L. ssp. indica plants were recorded in and around sugarcane fields at Shahjahanpur, Uttar Pradesh, India, during May-June 2013. Symptomatic plants yielded phytoplasma DNA bands of similar to 1.2 kb from nested PCR assays using universal phytoplasma primers P1/P6 and R16F2n/R16R2. BLASTn search analysis of both C. sativa ssp. phytoplasma isolates showed that the 16S rDNA sequences shared a 99 % of sequence identity between each other and with those of the phytoplasma strains of 16SrXIV group ('Candidatus Phytoplasma cynodontis'). Phylogenetic analysis of the 16SrDNA sequences of both phytoplasma isolates revealed their closest relationship with members of the 16SrXIV group. This supports the association of phytoplasma strains of the 16Sr XIV group with Cannabis little leaf and witches' broom diseases (Can LL-WB). Actual and virtual RFLP analysis of the 16S rDNA sequences of Can LL-WB phytoplasma isolates confirmed that they belonged to the 16Sr XIV-A subgroup. This is the first world report of a 16SXIV-A subgroup phytoplasma infection in C. sativa sp. sativa and C. sativa sp. indica plants.