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| Occurrence and Molecular Characterization of Groundnut Ringspot Virus (GRSV) Infecting Groundnuts in Western Kenya
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L W Murere, B Mukoye, M Kollenberg and H K Were
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Abstract:
Groundnut (Arachis hypogea) is an oilseed legume crop grown in Kenya on a small scale for income and nutritive value. However, its productivity remains low than its genetic potential of 1690 kg ha-1 attributed to biotic stressors. Viruses are among biotic stressors for low productivity globally. These include; Groundnut ringspot virus (GRSV), Tomato spotted wilt virus (TSWV),) among others. Groundnut ringspot virus had been reported in S.A, Ghana among others infecting groundnuts, peppers, watermelon, soybeans and many others. GRSV and TSWV have similar biological symptoms but differentiated serologically or by using molecular tests. Groundnut ringspot virus symptoms have been observed on groundnuts and other crops of economic importance in Kenya but no report had been documented on its occurrence or distribution. The objective was to Sequence nucleoprotein (N) genes of GRSV isolates obtained in a survey in western Kenya and compare with reference strains available in the GenBank. Plant GRSV Symptomatic samples were collected in a survey conducted in western Kenya and tested for GRSV using polyclonal antisera against GRSV. Samples that tested positive by DAS-ELISA was subjected to molecular tests. Total RNA was extracted from positive samples using CTAB and purified by DCC(tm)-5 purification kit then using target primers GRSVnR (5'-GCGGTCTACAGTGTTGCACTT-3') and GRSVnF (5'TCTTGTGCATCATCCATTGT-3') used to amplify the nucleoproteins (N) genes, at 614-bp fragment of the nucleocapsid gene of GRSV corresponding to the part of the nucleocapsid (N) gene using RT-PCR tests. The PCR product sequenced. Sequence readings trimmed using Bio-edit software and phylogenetic analysis done in MEGA-X. Kenyan GRSV isolates clustered with Brazilian, USA, Argentinian, Ghanaian and South African isolates in GenBank. The study showed that GRSV occurs in surveyed counties of western Kenya, which is a big concern. Introgression of resistant genes into local groundnut varieties be done with urgency to come up with varieties resistant to GRSV
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