ABSTRACT
Groundnut (Arachis hypogaea L.) is an economically important edible oilseed legume
in Sub-Saharan Africa (SSA). Smallholder farmers, who account for 75% of
producers, depend on it for food and income. However the yields are far below the
world averages. Groundnut rosette disease (GRD) is a major constraint of groundnuts
in Sub-Saharan Africa (SSA) causing up to 100% yield losses. The disease is caused
by two synergistic viruses; groundnut rosette assistor virus (GRAV, genus Luteovirus)
and groundnut rosette virus (GRV, genus Umbravirus) associated with a satelliteribonucleic
acid (Sat-RNA). Some of the setbacks in the epidemiological studies of
GRD associated viruses include the complex etiology of the disease and lack of
specific diagnostic tools. Simultaneous detection of the causal agents is possible by
multiplex RT-PCR but this depends on the availability of specific primers to known
agents that occur in a specific area. Information on occurrence and distribution of
GRD in western Kenya was not documented and little was known about the
characteristics of associated viruses. This study determined the distribution and
characterized GRD associated viruses in western Kenya. Two surveys were conducted
(2016/2017) in six counties; Bungoma, Busia, Homabay, Kakamega, Siaya and
Vihiga. Symptomatic and asymptomatic groundnut and some bean leafy samples were
collected for laboratory analysis. Total RNA was extracted from the leaf samples
using RNeasy Mini Kit (Qiagen) according to the manufacturers’ instructions and
used for double stranded cDNA synthesis using the SuperScript II kit. The cDNA was
column-purified with the DNA Clean & ConcentratorTM-5 – DNA kit. The samples
were then processed with the transposon-based chemistry library preparation kit
(Nextera XT, Illumina) following manufacturer’s instructions. The fragment sizes
structure of the DNA libraries was assessed using the Agilent 2100 Bioanalyzer. The
indexed denatured DNA libraries were sequenced (200-bp paired-end sequencing) on
the Illumina MiSeq platform (Illumina). Reads quality check was done using FastQC.
Trimmed reads were used for denovo assembly and contigs aligned to the viral
genomes database using CLC Genomics Workbench 10.1.2. The assembled contigs
were subjected to a BLASTn search against the GenBank database. Phylogenetic
analyses and comparisons were performed using MEGA X. Primers were designed
using Primer3Plus from consensus sequences. Biological characterization of GRD
was done through sap inoculation on leguminous hosts. Average incidence was 53%
and 41% in the short and long rain seasons, respectively. Chlorotic rosette was the
dominant symptom followed by Green rosette and Mosaic. Most farmers (65%)
sourced groundnut seeds from open air market. Complete nucleotide sequences of
Sat-RNA revealed identities of 88-100% with those from Malawi, Nigeria and Ghana.
Isolate EG16-5 clustered together with chlorotic M24S, all chlorotic isolates and
yellow blotch. The GRV isolates shared 84-98% sequence identity with those
available GeneBank. The GRAV coat protein (GRAV-CP) gene sequences revealed
97-100% identity with GeneBank isolates. Complete GRAV sequences clustered
closest with Luteoviruses in phylogenetic analysis. Leguminous hosts showed varied
symptoms and tested positive for Sat-RNA and GRAV using the designed primers.
The variations of GRD symptoms observed on groundnuts were due to the existence
of different variants of Sat-RNA. Sat-RNA and GRV are more diverse than GRAV.
The GRD viruses have hosts among the commonly grown legumes and this enhance
the perpetuation of the disease. The study recommends an urgent need to curb GRD,
possibly through the exploitation of pathogen derived resistance (PDR).
, M & , B (2021). Distributionand Characterization Of Groundnut Rosette Associated Virusesin Western Kenya. Afribary. Retrieved from https://tracking.afribary.com/works/distributionand-characterization-of-groundnut-rosette-associated-virusesin-western-kenya
, Mukoye and Benard "Distributionand Characterization Of Groundnut Rosette Associated Virusesin Western Kenya" Afribary. Afribary, 07 May. 2021, https://tracking.afribary.com/works/distributionand-characterization-of-groundnut-rosette-associated-virusesin-western-kenya. Accessed 22 Nov. 2024.
, Mukoye, Benard . "Distributionand Characterization Of Groundnut Rosette Associated Virusesin Western Kenya". Afribary, Afribary, 07 May. 2021. Web. 22 Nov. 2024. < https://tracking.afribary.com/works/distributionand-characterization-of-groundnut-rosette-associated-virusesin-western-kenya >.
, Mukoye and , Benard . "Distributionand Characterization Of Groundnut Rosette Associated Virusesin Western Kenya" Afribary (2021). Accessed November 22, 2024. https://tracking.afribary.com/works/distributionand-characterization-of-groundnut-rosette-associated-virusesin-western-kenya