Of 70 plant species tested, 50 species were susceptible to Tobacco streak virus (TSV) on sap inoculation. Both localized (necrotic and chlorotic spots) and systemic (necrotic spots, axillary shoot proliferation, stunting, total necrosis and wilt) symptoms are observed by majority of plant species. Eleven new experimental hosts were identified viz., Amaranthus blitum var. oleracea (Chaulai sag), Celosia cristata (Cocks comb), Beta vulgaris var. bengalensis (Palak/Indian spinach), Calendula officinalis (Pot marigold), Chrysanthemum indicum, Cosmos sulphurens (Yellow cosmos), Citrullus lunatus (Watermelon), Lagenaria siceraria (Bottle gourd), Coriandrum sativum (Coriander), Hibiscus subderiffa var. subderiffa (Roselle) and Portulaca oleraceae (Little hogweed). Detected groundnut seed infection with TSV for the first time by Direct antigen coated immunosorbent assay (DAC-ELISA) using whole seed. The seed infection ranged from 18.9 to 28.9% among the seeds collected from naturally infected and sap inoculated groundnut varieties (JL 24, TMV 2, Prasuna, Kadiri 6, Kadiri 9, Anantha and Kadiri 7 Bold) belonging to spanish and virginia types. Further, TSV was detected both in pod shell and seed testa and none of the samples showed the presence of TSV either in cotyledon or embryo. Grow-out and bio-assay tests proved the absence of seed transmission in groundnut and other legume crops. Hence, TSV isolate was not a true seed transmission case under Indian conditions in legumes.

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Tobacco streak virus (TSV, genus Ilarvirus family Bromoviridae) is known to cause stem necrosis disease (SND) in groundnut (Arachis hypogaea) since 2000 in Southern India. The TSV isolate infecting groundnut so far has not been characterized based on the complete genome sequence. In this study, TSV was isolated from a naturally infecting groundnut plant in Kadiri, the hot-spot of the SND in southern India. During the Kharif season of 2014, groundnut plants in an experimental field were affected with chlorosis and necrosis in leaf, stem and buds. The cent percent of the 48 samples with these symptoms collected from the field tested positive for TSV in ELISA samples in this context. One isolate, GN-Kad was established from a single lesion on cowpea cv. C-152 through successive sap inoculation. Cloning and sequencing of coat protein gene (717 nucleotides) of the isolate showed high sequence identity (98-99%) with the TSV isolates reported from different crops in India. The isolate produced local necrotic rings or veinal necrosis following sap inoculation to cowpea (cultivars C-152, Pusa Komal, Pusa Sukomal and Krishi Kanchan), French bean and sunflower; whereas, it produced systemic chlorotic mottling symptoms in Nicotiana benthamiana. The three segments of the virus genome (RNA 1, RNA 2 and RNA 3) contained 3523, 2903 and 2232 nucleotides, respectively. The overall genome sequence (8639 nt) of the present isolate shared 77-99% of nucleotide sequence identity with that of the other seven isolates reported from Australia, India and USA. The GN-Kad shared very close phylogenetic relationship with the okra and pumpkin isolates reported from India. The present report is the first comprehensive study of the molecular characterization of TSV associated with the stem necrosis disease of groundnut.Keywords Stem necrosis disease Á Groundnut Á Tobacco streak virus Á Complete genome Á India Electronic supplementary material The online version of this article (

Kpnl digestion. The fragments were cloned in pUC19 and sequenced. The partial sequences of four isolates were >95% idenfical to each other at the nucleotide (nt) level and thus only one isolate (P-25) was fully sequenced, determined to be 2,572 nt in length, and its sequence deposited in Gen-Bank (KFl 11683). The P-25 sequence showed a genome organization typical of a mastrevirus, with four open reading frames (ORFs), two in virion-sense and two in complementary-sense. The vidon and complementary-sense ORFs were separated by a long intergenic region, containing a predicted hairpin structure with the nonanucleotide sequence (TAATAT TAC) in the loop, and a short intergenic region. An initial comparison to all sequences in the NCBI database using BlastN showed the isolate to have the highest level of sequence identity with isolates of the dicot-infecting mastrevirus Chickpea chlorotic dwarf virus (CpCDV). Subsequent alignments of all available CpCDV isolates using the species demarcation tool (2) showed the isolate P-25 to share between 83.6 and 90.3% idendty to isolates of CpCDV available in databases, with the highest (90.3%) to CpCDV strain A odginafing from Syda (FR687959) (3). Amino acid sequence compadson showed that the predicted proteins encoded by the four ORFs of P-25 (coat protein [CP], movement protein [MP], replication associated protein A [RepA], and RepB) share 91.5, 88.2, 89.1, and 90.8% amino acid sequence identity, respectively, with the homologous proteins of the CpCDV isolate from Syda. Based on the recently revised mastreviruses species and strain demarcation criteria (78 and 94% whole genome nt identity, respectively) proposed by Muhire et al. (2), the results indicate that isolate P-25 represents a newly identified strain (strain F) of CpCDV. The pre.çence of CpCDV in symptomatic pepper plants was further confirmed by Southem blot hybddization technique using digoxygenin (DIG) labeled probe prepared from CpCDV isolate P-25. The genus Mastrevirus consists of geminiviruses with single component genomes that are transmitted by leafhoppers. Mastreviruses have so far only been identified in the Old World and infect either monocotyledonous or dicotyledonous plants (1). To our knowledge, this is the first report of a mastrevirus on the Arabian Peninsula and the first record of pepper as host of CpCDV. Recently, several begomoviruses of diverse geographic origins have been found infecting vegetable crops in Oman. The propensity of geminiviruses to evolve through recombination may lead to evolution of recombinant CpCDV with new host adaptability. Due to extensive agricultural/travel links of Oman with rest of the world, there exists high probability for the spread of this virus.

Reaction of five spanish (JL 24, TMV 2, Kadiri 6, Kadiri 9 and Anantha) and a virginia (Kadiri 7 Bold) groundnut cultivars was studied against Tobacco streak virus (TSV) by sap inoculation using different age group of plants [7-84 days after sowing (DAS)]. Among different cultivars, incubation period varied from 4 to 28 days and high incubation period recorded in Kadiri 7 Bold. The percent infection decreased with increase in the age of the plants in all the cultivars as it ranged up to 100 % in both pre-flowering (7-21 DAS) and flowering stage (28-63 DAS) plants followed by 50-100 % in maturity stage plants (70-84 DAS) except Kadiri 7 Bold. Similarly, 100 % wilting was observed in pre flowering stage plants against no wilting in maturity stage plants. However, cultivars differed in per cent wilting of flowering stage plants by recording maximum wilting (100 %) in JL24, Kadiri 6 and minimum (25.0 %) in Kadiri 7 Bold. Both localized (necrotic spots, veinal necrosis) and systemic (petiole necrosis, necrotic spots on young leaves, top growing bud and stem necrosis, axillary shoot proliferation, stunting, peg necrosis, pod necrosis, wilting of plant) symptoms induced by TSV were similar among all cultivars without any new symptoms. Prolonged stage of axillary shoot proliferation was observed for the first time in all the cultivars. In maturity stage plants of Kadiri 7 Bold, Kadiri 9 and Anantha, systemic symptoms restricted to leaf and petiole necrosis only. Virus titer varied significantly with the age of plants and inoculum harvest at days post inoculation and least virus titer recorded by Kadiri 7 Bold at all stages of infection. Among different cultivars, Kadiri 7 Bold was least susceptible/tolerant to TSV by registering higher incubation period with less per cent infection, wilt and titer.