Research outputs – some highlights
You can read about our full portfolio of 20 pump-prime funded research projects here. Below we have highlighted some of the excellent outputs from this work, all of which have made a difference to tackling vector-borne plant viruses in a range of important crops.
A further output is that the researchers, and members of the CONNECTED Network team, have produced around 25 publications.
A full-length clone of aphid lethal paralysis virus
As part of research project PPF#30 entitled ‘The prevalence of plant-vectored insect-pathogenic viruses in Kenya and their potential for bio-control of plant virus vectors’ a full-length clone of aphid lethal paralysis virus has been produced.
Farmers guidelines
As an outcome of CONNECTED research project PPF#11 entitled ‘Role of poleroviruses in maize lethal necrosis epidemics in Africa, a case study of Kenya’ revised farmers guidelines have been produced to include management of poleroviruses in the field.
Record of new aphid and its associated disease officially reported to Ghana’s National Plant Protection Organisation (NPPO)
As a output of CONNECTED research project PPF#31 entitled ‘Formal identification and characterisation of the viral pathogen causing a new aphid-associated disease of cabbage in Ghana’ a new aphid record and the plant disease carried by this aphid have been officially reported to Ghana’s National Plant Protection Organisation NPPO.
New Recombinase Polymerase Amplification (RPA) probe
As part of PPF#49 research project entitled ‘Rapid and inexpensive diagnostic tools for virus disease surveillance in sub-Saharan Africa’ a new RPA probe was developed.
This will be used in follow-up work to enhance the local capacity to produce diagnostic assays locally in Nigeria
Improvements to research infrastructure: First licensed entomopathogenic fungi (EPF) lab in Nigeria
As part of PPF#5 research project early career researchers developed new specialist skills in the biology of EPF and their evaluation as biopesticides. Insect pathology laboratories were established at IITA and Wesley University. Dialogue was established with the Nigerian plant health authorities over imports of EPF strains, and following inspection of facilities, approval was granted to hold imported EPF strains under licence. To our knowledge, the facility established at Wesley University is the first licenced EPF laboratory in Nigeria.
The establishment of this lab facility will allow researchers at Wesley University to further explore the use of entomopathogens as an essential component of integrated pest management (IPM) of cassava whiteflies.
RNA extraction method for cassava root material
As part of PPF#23 research project entitled ‘Is there a correlation between root necrosis symptoms and virus titre in cassava brown streak disease-affected cassava roots?’ a method was developed for the extraction of good quality RNA from cassava roots.
PCR primers
As part of PPF#11 research project entitled ‘Role of poleroviruses in maize lethal necrosis epidemics in Africa, a case study of Kenya’ PCR primers were developed to study poleroviruses.
Poleroviruses are one component within the maize lethal necrosis disease complex (MLN). MLN causes serious economic losses in East Africa. Primers can be used to better detect poleroviruses within plant material. This will help researchers in disease monitoring and surveillance.
Mobile phone app for improved plant disease detection in the field
As part of PPF#11 research project a mobile app for detection of maize lethal necrosis (MLN) /AMLN is undergoing testing in the field. Database development via insertion of images showing different disease symptoms including MLN/AMLN is on-going. Trained the system to recognize/identify MLND in the field for better detection of the disease using digital imaging. Currently testing the mobile App efficiency in the system to improve the accuracy in the field and inclusion of farmers, extension staff and other stakeholders to be part of the system as beneficiaries.
A new diagnostic protocol was developed that specifically detects the episomal form of badnaviruses infecting yam.
As part of PPF#36 research project entitled ‘Identification of mealybug vectors involved in the transmission of Dioscorea bacilliform viruses (DBVs) infecting yam in northern Nigeria’ a new diagnostic protocol was developed which significantly improved the detection of episomal badnaviruses infecting yam. Additionally, DNA extraction and mtCO1 amplification protocols were optimized for mealybug DNA barcoding/identification.
It is expected that the employment of efficient DBV diagnostic tools will enable the identification of areas with low insect vector pressure and re-infection rates needed for quality yam seed multiplications.
Locally made insect clip cages developed for aphid/viral disease transmission experiment
As part of PPF#36 research project entitled ‘Identification of mealybug vectors involved in the transmission of Dioscorea bacilliform viruses (DBVs) infecting yam in northern Nigeria’ locally made insect clip cages were developed and used for aphid/viral disease transmission experiment.
Making this piece of experimental equipment locally, rather than having to rely on buying in equipment from elsewhere in the country (Nigeria) or importing it, means that researchers are better able to conduct research on the transmission of virus diseases by aphids.
Method for nanopore sequencing on plant and insect materials to identify plant and insect viruses
As part of PPF#30 research project entitled ‘The prevalence of ‘plant-vectored’ insect-pathogenic viruses in Kenya and their potential for bio-control of plant virus vectors’ researchers have established a method for Nanopore sequencing of cassava, yam sweet potato and associated whitefly and aphid vectors to identify plant and insect viruses. The protocol includes methods for quality control of extracted RNA and steps for subsequent handling. Also, the researchers have more sensitive primers to confirm the identities of plant and insect material. To validate the protocol, the researchers have used it to analyse insect (whiteflies and aphid) samples collected from fields in Cote d’Ivoire.
The protocol will be made available for use across the 10 Central and West African countries that are implementing the West African Virus Epidemiology (WAVE) project https://wave-center.org/
Sampling methods were developed to enable assessment of viral infection in plants within a field setting
As part of PPF #1 research project entitled ‘Unifying CBSV surveillance’ sampling methods were developed for on site assessment of cassava brown streak virus (CBSV).
CBSD is a devastating disease that causes substantial economic losses in Sub-Saharan Africa. This work will contribute to research and surveillance of this disease.
Creation and testing of virus infectious clones
As part of PPF#14 research project entitled ‘Characterization of Cassava brown streak virus (CBSV) vector specificity and the role of its coat protein DAG motif in vector transmission’ mutant infectious clones have been made and tested in tobacco plants.
Infectious clones are a tool that can be used to understand plant-insect-virus interactions. The clones are being further evaluated as part of a Bill and Melinda Gates Foundation (BMGF) funding award with the University of Greenwich.
Data set created as part of research project entitled ‘Tracking genetic diversity and development of diagnostic tools for maize lethal necrosis (MLN) viruses in Uganda’
Data set created as part of project to explore whether diverse virus combinations affecting maize causing MLN across major maize growing areas of Uganda. Project objectives: Obj. I To determine incidence and severity of MLN in Uganda Obj. II To sequence and identify uncharacterized viruses causing MLN in Uganda Obj. III To develop diagnostic protocols of uncharacterized viruses causing MLN
An international collaboration of researchers have joined forces to work with this data along with other HTS sequencing data obtained in CONNECTED PPF research projects.
Data set created as part of research project entitled ‘Rapid and inexpensive tools for the detection and Surveillance of viruses in insect vectors and hosts in Africa’
Dataset created as part of a project on the development of rapid inexpensive diagnostic tools for the detection and monitoring of vector borne viral plant disease infection in Sub Saharan Africa. Project objectives:
• To develop simple and rapid diagnostic tests usable on-site for the surveillance of established and emerging virus threats to major food and horticultural crops in sub-Saharan Africa (SSA)
• To develop rapid flow through assays for with a colorimetric readout for Maize chlorotic mottle virus (MCMV) and Maize Streak Virus (MSV)
• To validate recombinant polymerase assays (RPA) for Banana Bunchy Top Virus (BBTV) with Fluorescent (Exo kit, Twist-Dx) and lateral flow readouts (nfo kit Twist-Dx). and to compare commercially and laboratory produced lateral flow/flow through assays.
Data set created as part of research project entitled ‘Cucurbit viruses and their control in the smallholder farming communities of Zimbabwe’
Data set created as part of project that sought to address the hypothesis that pumpkin-infecting viruses are widespread and severe in Zimbabwe. Project objectives: 1. Assessment of the geographical distribution, incidence and severity of pumpkin-infecting viruses in Zimbabwe. 2. Characterization of the viruses infecting pumpkins in Zimbabwe. 3. Disseminating information about pumpkin viruses in Zimbabwe.
Full genome sequences of pumpkin-infecting viruses from Zimbabwe were generated and deposited in public databases. These can be used by Universities and research institutes to study mechanisms of pathogen evolution, development of ELISA test kits, breeding for disease resistance and development of other disease control strategies.
Data set created as part of research project entitled ‘Occurrence, distribution and molecular diversity of virus stressors on groundnut crops in Kenya’
Data set created as part of project to identify viruses that cause disease in the main groundnut producing areas of Kenya and use this information to develop IPM control strategies. For example, to control the main insect vectors, or avoid planting when and where vectors are active. Project objectives were: 1. To determine occurrence and distribution of groundnut viral diseases in main growing areas of Kenya
2. To build capacity of MMUST scientist in plant virus detection and identify viruses causing the diseases in objective 1 above
3. To develop, optimise or devise diagnostic tools for the viruses identified in Objective 2 above.
Data set created as part of research project entitled ‘Novel botanical formulations for treatment of virus vectors’
Data set created as part of a project to explore whether novel botanical formulations be used as interventions to reduce populations of virus vectors on crops and consequently prevent the spread of plant virus diseases. This included the following project objectives:
• Prepare a range of novel nanoformulations of plant essential oil extracts
• Conduct bioassays on the target virus vectors with novel nanotech formulations of plant secondary metabolites
• Conduct preliminary evaluation of novel nanotech formulations of plant secondary metabolites for use against virus vectors
• Determine which botanicals are widely used in Africa
Data set created as part of research project entitled ‘Identification of vectors involved in the transmission of Maize chlorotic mottle virus infecting maize in Eastern Africa’
One of the project objectives was to investigate the diversity of invertebrates in maize fields in locations with high and low Maize Lethal Necrosis incidence. This resulted in sequence data from Minion sequencing of insect material. Novel metabarcoding methods to identify insects, based on MinIon sequencing, were evaluated
An international collaboration of researchers have joined forces to work with this data along with other HTS sequencing data obtained in CONNECTED PPF research projects.
Personal career development
Dr KF, the PI on one of our PPF projects, has told us that his participation in the PPF project enabled him to successfully apply for internal promotion to the Position of Associate Professor of Entomology. Also that appointed as the coordinator of the West African sub-regional Centre of the African Regional Postgraduate Programme in Insect Science (ARPPIS), based at the University of Ghana, effective 1st October 2021