IPVDN – Training in virus detection and
diagnosis, capacity building, and delivery of
IPM packages
7th International IPM Symposium
Memphis, Tennessee
March 27-29, 2012
Capacity building and short term training:
Requirements for successful technology transfer
for Integrated Pest Management
Sue A. Tolin, Virginia Tech
stolin@vt.edu
The IPM CRSP IPVDN Team – 2009-2014
Plant Virus Disease US Participants and Regions
Sue Tolin, Program Leader
Judy Brown
Naidu Rayapati
UAZ
WSU
Mike Deom
Bob Gilbertson
UGA
UC-D
South Asia
Latin America &
Caribbean
Honduras
Guatemala
Ecuador
West Africa
Mali
Senegal
Ghana
c
Central Asia
India
Bangladesh
Nepal
Tajikistan
Southeast Asia
East Africa
Uganda
Kenya
Tanzania
Indonesia
Philippines
Cambodia
IPM CRSP - International Plant
Virus Disease Network (IPVDN)
Objectives:
1. Define virus disease problems in key crops
through surveys and virus diagnosis and
detection
2. Build in-country capacity for research on
virus detection, ecology, and epidemiology
3. Develop and help deliver IPM packages for
virus management
Virus Diagnosis and Detection are needed
for Disease Management
 Diagnose virus and identify vector and sources
 With in-country capacity building and training
 Involve plant pathologists and entomologists
 Predict potential management practices
 Design experiments to assess success of interventions
 Provide models and approaches to be used by IPM
scientists and practitioners
 Technology transfer to introduce ecologically-
based management system practices
 Most developing countries lack this capacity !
Why do virus diagnosis?
 Curious to know the cause of a symptom
 One or more viruses? Phytoplasma? Fungi? Bacteria? Insect?
 Growers want an answer – What can I spray?
 To know what virus or viruses are present to develop a
management practice – virus IPM is long term plan
 Source? Where did the virus come from?
 Can the source be reduced? Weeds? Seeds?
 How is the virus transmitted?
 Biological – specific vectors transmit certain taxonomic
groups of viruses
 Mechanical – by human contact during transplant and
harvest is important for certain viruses
 Seed – Only common for certain taxonomic groups of
viruses
Why do virus detection?
 Monitor effectiveness of in-field management
 Vector control with pesticides
 Time of planting, barrier crops to avoid vectors
 Host-free periods to reduce virus in vector and weeds
 Help breeders develop virus resistant varieties
 Provide virus-tested planting material
 Citrus tristeza, banana viruses, potato, other
vegetatively propagated plants
 Seeds free of seed-transmitted viruses
 Test for phytosanitary regulations for export
 High sensitivity needed
Training and Capacity Building
 Workshops in host countries
 Honduras workshop on virus management
 As a part of IPDN workshops
 Separate virology workshops
 Short term visits by trainees to US
$
laboratories
 1-2 weeks to 6 months
 Enhanced capacity
 Graduate degree training
 Full-time; sandwich programs
 Needed for full capacity
$$
$$$$
Comayagua, Honduras workshop for field
extension workers from various projects
 PURPOSE: To transfer to field extension workers practice-
oriented (and some theoretical), up-to-date information on
management of virus diseases, focused on their
nature/biology, dissemination, transmission, epidemiological
aspects, e.g., influence of weather, genotypic differences,
cultural practices, and other factors. At the end, participants
should be able to understand the rationale of and knowingly
apply the recommended management measures to combat
viruses.
 1. BASIC PRINCIPLES OF PLANT VIROLOGY. Their
importance, nature, and characteristics of the viruses and
virus diseases of major relevance to their management.
 2. MANAGEMENT OF VIRUS DISEASES IN CROPS.
Strategies and management practices used in the open field
and protected environments before, during and after the
cultivation cycle.
Training Workshops - Lectures
1. The nature of viruses – what are they!
2. Types of symptoms caused by viruses
3. Virus ecology – plants infected, ecological
niches in ecosystems
4. Virus epidemiology – how do viruses get
to crop plants
 Via vectors?
From seed? Other ways?
5. Virus diagnosis and detection methods
6. Apply management and monitor results
Diagnostic Workshops (IPDN)
Univ del Valle de Guatemala – 1 day of Virus Training
 Hands-on methods
 Immunostrips
 Indirect ELISA
 Tissue blot immunoassay
 Inclusion bodies
 PCR for DNA viruses
 FTA Cards
Optimizing membrane-based technologies for virus identification
TBIA
RT-PCR + sequencing
Virus Diagnosis in Workshops
 Immunoassays – reaction of virus-specific
antibodies with virus particles
 ELISA – DAS, PTA – microtiter plates
 Tissue print or blot ELISA on membranes – TBIA
 Lateral flow devises on membranes - Immunostrips
 Molecular assays – detection of specific nucleic acid
sequences of viral genome
 Amplification by polymerase chain reaction (PCR)
 Nucleic acid recovery methods; Primer Design
 Reverse Transcription, Real-time
 New sequencing methods and analysis
Facts about Virus Diagnosis
 Virus can rarely be diagnosed by symptoms
 Strains, host and variety, environment, time of infection
all cause variations in symptoms
 Plants may have multiple viruses
 Diagnostic tests are available for only certain
known viruses or virus families
 There are no rapid diagnostic tests for unknown
viruses
 New viruses are emerging rapidly
 Or, viruses have just now been recognized and
characterized
Choosing a diagnostic/detection test?
Information, Needs, and Capacity
 What viruses are known in the crop?
 What tests are available for those viruses?
 What specificity and sensitivity are needed?
 Quarantine and clean planting stock - high
 Monitoring for IPM and disease management lower
 What tests can you conduct?
 Immunoassays? PCR? Biological? Physical?
 What tests can you afford to do?
Crop, Dissemination, Vector, Virus genus, Symptom
Eggplant
Pepper
Tomato
Mechanical, contact
Major insect vectors
Other vectors
• Nematode, fungi, mite,
grasshopper, beetle
Aphid
Potyvirus
Whitefly
Cucumovirus
Mottle / Mosaic
Thrips
Begomovirus
Chlorosis
Necrosis
Tospovirus
Potexvirus
Stunting / Distortion
Tobamovirus
Tombusvirus
Deformation of plant parts
India –
Tamil Nadu Agricultural University
 First virus workshop July 12-16,2010
 Week-long program, 1.5 day hands-on
exercises in virus detection/diagnosis, 1 day
field trip.
 Tolin and Naidu gave most lectures; some by
Indian and Indonesian scientists
 All participants spoke at beginning and end
of the workshop to introduce their own work,
and comment on what they learned.
Tamil Nadu Agricultural University, Coimbatore, India
Workshops for
capacity building
in plant virus
diseases
IPVDN Global Theme
July 2010
Participants:
South Asia –
India
Southeast Asia –
Cambodia,
Indonesia
Central Asia –
Uzbekistan
Asia Virus Workshop at TNAU:
Detection and Diagnosis: Hands-on Activities
Symptom Observation
Biological Detection
Mechanical Transmission
See results 4 days later
Asia Virus Workshop at TNAU:
Detection and Diagnosis: Hands-on Activities
Diagnostic Immunoassays
Classical ELISA
ELISA on Membranes
Plant Disease and Insect Pest Diagnostics Workshop –
Bogar, Indonesia July 22-23, 2010
 First day, lectures on use of serology and PCR in disease
diagnostics, and lecture and lab sessions on diagnosis of
virus
 Conducted by Drs. Tri Damiyanti and Sri Hendrastuti Hidajat,
who had attended the TNAU Workshop the week before
 They demonstrated Tissue Blot Immunoassay with
nitrocellulose membranes and reagents used by Tolin at
TNAU - and first applied in the IPM CRSP in Jamaica in
Caribbean Site, Phase II, before the Virus Global Themes
Central Asia
 IPDN Workshop held June 6-11, 2011 at Tajikistan
National University in Tajikistan (Naidu)
 A strong need for increased effort in capacity
building in diagnosis and management of virus
diseases
 Candidate from Tajikistan will be sent for short
term training at Naidu’s laboratory in potato virus
detection and seed certification
Planned Activities
 IPDN/PVD workshop in May 2012 in
Tanzania to train using SOPs for diagnosis –
includes 3-4 viruses
 Workshop in India – TNAU on Virology
 July 13-16, 2012
 Degree training
 Washington State University
 Univ California – Davis
 Honduran MS student to begin - Arizona