Insect Vectored Disease Workshop, GPDN

Everyone is invited to attend the GPDN business meeting and No host dinner after the meeting . The first session is Tuesday with a follow up session Friday morning.

Workshop participants arrive in Bozeman, Best Western City Center

GPDN Dinner @ 6:00 pm at a fine dining establishment (TBA) somewhere on Main street east of the City Center Motel.

Wednesday, September 28

Morning, Day 1 – 8:00 to 12:00

Welcome and Introduction (J. Riesselman)

Techniques for preparing plant disease vectors (mites, aphids) for remote ID or mailing (Mike Ivie, MSU)

Also discussion about similar procedure to preparing leafhoppers (A. Wayadande)

Hands-on sampling mite colony for specimens.

Slide preparation of mites, cooking show style.

Demonstration and note making.

30 minutes

Mites

Cleaning

Ultrasconic cleaners

10 minutes

Clearing

Lactic acid

Hot Plates

20 minutes to overnight

Specimen drying

ETOH, Orange oil

Hood

2 hours

Embeding

Hoyers

10 minutes

Leafhoppers

Cleaning

KOH or acetic acid

Clearing

small glass vials, glass slides

Specimen drying

Mounting

permount or other mounting medium

good forceps, razor blades, sharpie felt-tip fine point pens

Slide Making

Continue Afternoon, Day 2 - 3:00 to 5:00

Afternoon, Day 1 – 1:00 to 5:00

Slide preparation for identification of leafhoppers to genus/species: (A. Wayadande)

1. cleared male genitalia

2. female seventh abdominal sternite

3. wing venation

4. characteristic frons/head/pronotum markings

Demonstration of cleared male genitalia (samples) and sample prep

30 min

Slide preparation

10 min

Female 7^th and wing venation prep

20 min

Resources for leafhopper/planthopper identification

5-10 min

Discussion:

10 - 15 important leafhopper species that vector disease pathogens

60 min

disease symptoms, leafhopper vector species, some biology (migratory behavior, habits, oviposition stuff that diagnosticians might find helpful)

Whiteflies and thrips and molusks. Supported by handouts.

15 min

Diagnosis of disease (bacteria, mollicutes)

Diene’s stain sample prep and examination

30 min

Visualize molicutes, spiro and phytoplasma

Equipment and supplies: compound microscope, glass slides, Diene’s stain (I’ll bring it or we can make it there, razor blades, mollicute-infected plants and healthy controls

Leafhopper Vectors of Plant Pathogens

Thrips vectors of tomato spotted wilt two spot virus

Evening, Day 1 – 6:00

Social event prepared by Jack Riesselman, roaming pathologist and connoisseur extraordinaire.

Menu:

Seared Montana London Broil with Green Peppercorn Sauce

Grilled Poached Champagne Dilled Salmon

Hutterite Riblets with Madison Junction BBQ sauce*

Clark Fork Valley Baked Beans

Manhattan Dutch Potato Salad

Celery Seed Cole Slaw Vinegarette Salad

Yellowstone Valley Athena Sweet Cantaloupe with Prarie Wildflower Honey.

Entertainment: Ringling 5, Montana's answer to well you decide… http://www.ringling5.com/main.htm

RickRanchers Leg.JPG

* Bring your favourite napkin to tuck into your collar because Hutterite riblets require focus. Distractions might allow the locals to gain the advantage.

Thursday, September 29

Morning, Day 2 – 8:00 to 12:00 Important Insect Vectored Viruses and Methods for Diagnosis

8AM – ELISA (Nina Zidack and Courtney Pariera-Dinkins)

Principles

Sources for kits and reagents

Applications in our lab

Demo of ELISA

Principles and application of dip strips

Demo of dip strips

Elisa Basics

Immuno Strips.pdf

Dalmatian Toadflax (Linaria dalmatica).pdf

9AM – PCR (Eric Gillitzer)

Principles

Sources for primers

Demo of PCR

Story about diagnosing brome mosaic virus

PCR applications for detecting viruses in insects – Astri Wayadande

PCR

10AM – Break

10:30 – Diagnostics for Virus diseases of wheat (Wheat Streak Mosaic, Barley Yellow Dwarf Mosaic Virus, High Plains Virus) – Mark Young

Barley Yellow Streak Mosaic Virus – Sue Brumfield

Observation of brown wheat mites and wheat curl mites

Mt Viruses Talk

BaYSMV

Time Remaining – Discussion of individual labs diagnostic techniques

12:00 AM Lunch on Your Own

1:00 PM – Use of Transmission Electron Microscopy for diagnosis of plant viruses – Sue Brumfield (Meet at 108 ABS, go to 227 and 230 Leon Johnson Hall)

ISEM and negative stain techniques

Continued from Morning, Day 1 - 3:00 to 5:00

Techniques for preparing plant disease vectors (mites, aphids) for remote ID or mailing (Mike Ivie, MSU)

Labeling

Pens and labels

20 minutes

Curing

Oven

overnight

Ringing

Ringer, Paint brushes

10 minutes

Short slide evaluation

Snapdragon Demonstration (Will Lanier, Rex Hoover)

Snapdragon, TriTek Labs

60 minutes

Lucid Key

Lucid key description

30 minutes

Slide Preperation Lab Take home:

Get the bubbles out from under a mite and control the sticky stuff (mounting fluid).

Solve the swirl problem, position specimen and understand voucher and standard process.

1 ID slide each and manual

How to get an identification of your slide

Current - Remote diagnosis via the GPDN using Web microscope, digital scanner and cameras.

Future - Snapdragon Demonstration - Remote diagnosis via 5 axis rotational imaging equipment. (R. Hoover / W. Lanier)

Sampling of area agriculture systems with related MSU faculty. Post Research Farm, Northern High Plains (Dry and High) region cropping systems.

Tour of MSU resources like the Plant Growth Center and Quarantine, New AgBioScience facility, Mycology, Museum of the Rockies.

Bag - a - Peak, Hike to the "M"

Friday, September 30

Morning, GPDN Business meeting, Room 326 Leon Johnson Hall

Workshop participants depart.