Govind P RaoSHREENATH Y S2019-02-132019-02-132018http://krishikosh.egranth.ac.in/handle/1/5810094825t-9936Severe stunting, little leaf and excessive axillary shoot proliferation symptoms were observed in chickpea filed at IARI, New Delhi, Udaipur and Jhansi in January-March 2017-18 with disease incidence ranging from 6-56%. The association of Ca. P. trifolii subgroup D phytoplasma was confirmed only in symptomatic chickpea stunt plants of variety Pusa Bheema at experimental field of Division of Entomology of IARI, New Delhi in direct and nested PCR assays with phytoplasma-specific universal primer pairs and in silico RFLP analysis. Leafhopper species, Hishimonas phycitis, feeding on chickpea symptomatic plants at IARI, Delhi fields were also found positive for association of 16SrVI-D phytoplasma subgroup. Seven symptomatic chickpea collected samples (two from Delhi, three from Jhansi and two from Udaipur) were tested in leaf dip preparation under electron microscope and showed presence of spherical icosahedral virus particle of 25-30 nm diameter in chickpea stunt Delhi sample-1 and the aphid, Myzus persicae, which feeds on the symptomatic chickpea at IARI, New Delhi. The CpS Delhi sample-1 showed positive serological relationship with polyclonal antibody of CMV in DAS-ELISA and decoration test. All the seven CpS samples from three states namely New Delhi, Rajasthan, Uttar Pradesh were tested positive for CpCDV and showed 99% sequence identity with Chickpea chlorotic dwarf Pakistan virus confirming the presence of CpCDV. RCA was also performed with CpS Delhi sample-1 and ~ 2.7 kb amplified sequence identified as CpCDV strain from Pakistan. No luteovirus specific amplification was achieved by utilizing Luteoviridae specific primers in any of the seven chickpea stunt samples indicating the absence of luteoviruses. However, association of CMV was confirmed with CpS Delhi sample -1 and Myzus persicae collected from chickpea fields at IARI, with CMV coat protein specific primer pairs. The results of study suggested a mixed infection of Candidatus Phytoplasma trifolii (16SVI-D subgroup), CMV and CpCDV (Mastrevirus) associated with CpS sample-1 from IARI, New Delhi and association of only Mastrevirus with rest of the chickpea stunt samples collected from Delhi, Jhansi and Udaipur. This is the first report of mixed infection of virus and phytoplasma associated with chickpea stunt disease and the association of Candidatus Phytoplasma trifolii (16S VI-D subgroup) with chickpea stunt disease from the world. Keywords : Mastrevirus, Candidatus Phytoplasma trifolii , Hishimonas phycitis, Myzus persicae, Cucumovirusen-USnullThesis