First report of a ‘<i>Candidatus</i> Phytoplasma aurantifolia’ (16SrII‐D) isolate associated with little leaf and stunting of <i>Dracaena reflexa</i> in India

Dracaena reflexa commonly known as “song of India” is an important ornamental plant grown both outdoors and a house plant. During October 2021, typical phytoplasma symptoms including little leaf stunting were observed in D. plants growing the landscape around Dr.Y.S.R. Horticultural University, Anantharajupeta, Andhra Pradesh. Eight diseased amongst 100 surveyed (Figure 1). Total DNA was extracted from samples collected two one symptomless using CTAB method (Murray & Thompson, 1980). extracts subjected to nested PCR universal phytoplasma-specific primer pairs P1/P7 (Deng Hiruki, 1991) for direct round R16F2n/R16R2 amplification 16SrRNA gene (Gundersen Lee, 1996). Primers rp(II)F/rp(I)R1A followed by semi-rp(II)F2/ rp(I)R1A) used ribosomal proteins (rp gene) (Martini et al., 2004). Amplicons expected size (c. 1.25 Kb 16S rRNA c. 1.29 rp obtained but not samples. products cloned (pGEM-T Easy Vector; Promega, USA) directly sequenced (ABA Biotech, India). Sequences deposited GenBank with Accession Nos. ON787808 ON787809 (16SrRNA ON791584 ON791585 gene). BLASTn analysis showed that partial 16Sr RNA sequences isolates shared 100% identity ‘Candidatus Phytoplasma aurantifolia’ (MT420677, MT420671). The sequence identities ranged 99.92-100% compared ‘Ca. P. aurantifolia’, 16SrII group (EF193377, EF193371). A sequence-based phylogenetic analyses 2) associated clustered strains. virtual RFLP based on performed iPhyclassifier (https://plantpathology.ba.ars.usda.gov) which revealed pattern identical phytoplasmas II, subgroup D (Y10097) similarity coefficient 1.00. To best our knowledge, this first report 16SrII-D infection reflexa. (16SrII-D subgroup) strains have been reported commercial crops, weed hosts several leafhopper vectors India (Reddy 2021). This indicates related (16SrII-D) are widespread wide host range India. Further research into epidemiology may provide useful information implement effective management recommendations. authors would like thank Associate Director Research, Regional Agricultural Research Station, Tirupati support encouragement.