First Report of Tomato yellow ring virus (Tospovirus, Bunyaviridae) Infecting Tomato in Kenya

Tomato (Lycoperscion esculentum) is one of the most popular vegetables and a major source of nutrition and income for smallholders in Africa. Thrips-transmitted tospoviruses are among the economically important pathogens of tomatoes that cause significant crop losses worldwide (3). In surveys for Tomato spotted wilt virus (TSWV) in the major tomato production areas of Kenya between March 2010 and January 2012, tomato fruits with chlorotic ring spots on fruits with stem and leaf necrosis were observed frequently. The symptoms were more evident in the dry seasons and disease incidence ranged from 28 to 42%. The pathogen did not react with antiserum specific to TSWV (Agdia Biofords, Ervy, France) in double-antibody sandwich (DAS)-ELISA. Furthermore, the pathogen did not react with antiserum specific to Capsicum chlorosis virus (CaCV), Chrysanthemum stem necrosis virus (CSNV), Groundnut ring spot virus (GRSV), Impatiens necrotic spot virus (INSV), Iris yellow spot virus (IYSV), and Watermelon silver mottle virus (WSMoV) (Agdia Biofords and DSMZ, Germany) in DAS-ELISA, but reacted positively to antiserum specific to Tomato yellow fruit ring virus (TYFRV) (DSMZ, AS0526). The nucleocapsid (N) gene specific primers (TFfor: 5'-ACTCATTAAAATGCATCGTTCT-3' and TFrev: 5'-CTAAGTAAACACCATGGCTACC-3' as forward and reverse primers, respectively) were designed by choosing six conserved regions of the N gene sequences of known TYFRV and Tomato yellow ring virus (TYRV) sequences available from GenBank. Using these primers, TYRV infection of tomatoes collected from Loitokitok, Kenya (2.73°S, 37.51°E) was confirmed by reverse transcription (RT)-PCR. PCR products of approximately 912-bp were obtained from six out of 11 symptomatic tomato samples tested, but not from healthy and water controls. Amplicons were gel-purified using QuickClean II Gel Extraction Kit (GenScript, UK) and sequenced using TFfor and TFrev primers. A consensus sequence was generated using Geneious Pro 5.5.6 Software (Biomatters Ltd., Auckland, NZ). The BLAST revealed that the N-gene sequence of the Kenyan tomato isolate (GenBank Accession No. JQ955615) had sequence identity with the Cineraria isolate (98.5%) (Accession No. DQ788693.1) and the Anemone isolate (98.1%) (Accession No. DQ788694.1) of TYRV (4) from Fars Province, Iran; an Alstroemeria isolate (98.4%) (Accession No. HQ154130.1) and two tomato isolates (98.3%) (Accession Nos. HQ154131.1 and AY686718.1) of TYRV from northern Khorasan Province, Iran, a