Association of Apple necrotic mosaic virus (ApNMV) with mosaic disease in commercially grown cultivars of apple (Malus domestica Borkh) in India

During the course of survey, an incidence of 7.14-90% of apple mosaic disease (AMD) was recorded in apple orchards in Jammu and Kashmir among various commercially grown cultivars. The maximum incidence of mosaic disease was observed in cultivar Golden Delicious. In addition to mosaic, symptoms of chlorosis, necrosis and ring spots were also observed. In the present study association of Apple necrotic mosaic virus (ApNMV) was confirmed by RT-PCR and sequencing of whole coat protein gene in samples tested negative for Apple mosaic virus (ApMV) in DAS-ELISA. Out of 18 samples tested in RT-PCR, ten were found positive for ApNMV. Out of ten ApNMV positive samples, amplicon of 680 bp of samples representing five cultivars were sequenced and sequence analysis showed 89-91% sequence identity with ApNMV. The phylogenetic analysis grouped Indian isolates into two sub-clusters under one major cluster (ApNMV group). The sub-cluster-I, included ApNMV isolates from cultivars, Oregon Spur, Red Delicious and Fuji Aztec along with Chinese and Korean isolates. Sub-cluster-II included ApNMV isolates associated with Golden Delicious and Royal Delicious. The comparison of coat protein gene-based sequence identity matrix showed maximum and minimum similarity of 89-99% with ApNMV isolates from China. It also showed maximum identity with PNRSV (61.6%) and ApMV (52.8%) under subgroup 3 of genus Illarvirus. Our study indicates that the ApNMV is commonly associated with AMD in India and may be a major cause of the mosaic disease in apple cultivars. To the best of our knowledge, this is the first report of the association of ApNMV with apple mosaic disease from India.