7th International IPM SymposiumMemphis, TennesseeMarch 27-29, 2012

Capacity building and short term training: Requirements for successful technology transfer

for Integrated Pest Management

Sue A. Tolin, Virginia Techstolin@vt.edu

IPVDN – Training in virus detection and diagnosis, capacity building, and delivery of

IPM packages

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The IPM CRSP IPVDN Team – 2009-2014Plant Virus Disease US Participants and Regions

Judy BrownUAZ

Mike DeomUGA

Bob GilbertsonUC-D

Naidu RayapatiWSU

East AfricaUgandaKenya

Tanzania

Latin America & CaribbeanHonduras

GuatemalaEcuador

West AfricaMali

SenegalGhana

Central Asia

Tajikistan

South AsiaIndia

BangladeshNepal

Southeast AsiaIndonesiaPhilippinesCambodia

Sue Tolin, Program Leader

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 sourcesWith in-country capacity building and trainingInvolve plant pathologists and entomologists

Predict potential management practices Design experiments to assess success of interventionsProvide 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 planSource? 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 managementVector control with pesticides Time of planting, barrier crops to avoid vectorsHost-free periods to reduce virus in vector and weeds

Help breeders develop virus resistant varietiesProvide virus-tested planting material

Citrus tristeza, banana viruses, potato, other vegetatively propagated plants

Seeds free of seed-transmitted virusesTest for phytosanitary regulations for export

High sensitivity needed

Training and Capacity BuildingWorkshops in host countries

Honduras workshop on virus managementAs a part of IPDN workshopsSeparate virology workshops

Short term visits by trainees to US laboratories 1-2 weeks to 6 monthsEnhanced capacity

Graduate degree trainingFull-time; sandwich programsNeeded 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 viruses3. Virus ecology – plants infected, ecological

niches in ecosystems4. Virus epidemiology – how do viruses get

to crop plants Via vectors? From seed? Other ways?

5. Virus diagnosis and detection methods6. Apply management and monitor results

Diagnostic Workshops (IPDN)

Hands-on methodsImmunostripsIndirect ELISATissue blot immunoassay

Inclusion bodiesPCR for DNA virusesFTA Cards

Univ del Valle de Guatemala – 1 day of Virus Training

TBIA RT-PCR + sequencing

Optimizing membrane-based technologies for virus identification

Virus Diagnosis in Workshops

Immunoassays – reaction of virus-specific antibodies with virus particles ELISA – DAS, PTA – microtiter platesTissue print or blot ELISA on membranes – TBIALateral flow devises on membranes - Immunostrips

Molecular assays – detection of specific nucleic acid sequences of viral genomeAmplification by polymerase chain reaction (PCR)

Nucleic acid recovery methods; Primer DesignReverse Transcription, Real-timeNew sequencing methods and analysis

Facts about Virus Diagnosis

Virus can rarely be diagnosed by symptomsStrains, host and variety, environment, time of infection

all cause variations in symptomsPlants may have multiple virusesDiagnostic tests are available for only certain

known viruses or virus familiesThere are no rapid diagnostic tests for unknown

virusesNew viruses are emerging rapidlyOr, 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 - highMonitoring for IPM and disease management -

lowerWhat tests can you conduct?

Immunoassays? PCR? Biological? Physical?What tests can you afford to do?

Crop, Dissemination, Vector, Virus genus, Symptom

Tomato Pepper Eggplant

Major insect vectors Mechanical, contact

TobamovirusCucumovirus PotexvirusPotyvirus Begomovirus

Other vectors• Nematode, fungi, mite,

grasshopper, beetle

Tospovirus Tombusvirus

Mottle / Mosaic Chlorosis Necrosis Stunting / Distortion Deformation of plant parts

Aphid Whitefly Thrips

India – Tamil Nadu Agricultural University

First virus workshop July 12-16,2010Week-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.

Workshops for capacity building

in plant virus diseases

IPVDN Global ThemeJuly 2010

Tamil Nadu Agricultural University, Coimbatore, India

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 AsiaIPDN 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 ActivitiesIPDN/PVD workshop in May 2012 in

Tanzania to train using SOPs for diagnosis – includes 3-4 viruses

Workshop in India – TNAU on VirologyJuly 13-16, 2012

Degree trainingWashington State UniversityUniv California – DavisHonduran MS student to begin - Arizona