In the course of the winters of 2018 to 2020, witches’ broom, phyllody, flat stem, little leaf, yellowing and stunting symptoms have been recorded in Mexican and French marigold fields in 4 states of India. The average recorded illness incidence ranged from three to 30% in numerous states. 1.2 kb product of 16S rRNA gene was amplified in symptomatic marigold samples utilizing common phytoplasma-specific nested primer pairs. Sequence comparison, phylogenetic and virtual RFLP analysis of 16S rRNA gene sequences identified the phytoplasmas in marigold samples as enclosed in the 16SrII-C subgroup for the strains from Delhi and within the 16SrI-B subgroup for the strains from Tripura, Uttar Pradesh and Maharashtra. Phytoplasma presence was further confirmed amplifying 4 genes (secA, rp, secY and tuf) in the symptomatic marigold species from all the surveyed places. Moreover, six weed species rising in marigold fields resulted positive for phytoplasmas. Phyllanthus niruri (Delhi) was examined constructive for 16SrII-C subgroup phytoplasma, nonetheless, Ageratum conyzoides, Acmella calva and Ludwigia perennis (Tripura), Cannabis sativa (Uttar Pradesh) and Parthenium hysterophorus (Maharashtra) were tested positive with 16SrI-B subgroup associated phytoplasma strains. Four leafhopper species were additionally detected constructive for 16SrI-B subgroup collected from marigold fields at Tripura, Uttar Pradesh and Maharashtra and have been recognized as putative vectors for identified marigold phytoplasma strains.
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