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Inoculation Technique

Technique

Like most of the procedures in this business, the process of inoculation is highly dependent on weather. Keep your eye on this website for weather updates. If rain is forecast for an entire scheduled inoculation day, inoculations will be held the following day (see rain date); however, if it looks like the rain will be spotty, we'll wait until passing showers finish their business. Should the forecast call for extended periods of cold and rain following the inoculation day, likewise, inoculations will have to be postponed. Keep your fingers crossed for fair weather.

1. Organize team of four to six - one plugger, one to two inoculaters, one to two tapers, and a recorder.    Prepare supplies for each team member.

Plugger - sanitizes cork bores by flaming with 190-proof grain alcohol
(Warning - don’t stick bore barrel into alcohol too deeply or you will end
up with a blow torch)(Beware - you can’t see the blue flame, but you can feel it); Also prepares wipe rag with 70% ethanol alcohol to sanitize areas of trunk to be inoculated, and prepares caddy with test tube secured and filled with 190-proof alcohol, extra alcohols, 2 lighters and rags.

Inoculators- each prepares caddy for alcohol bottle, test tube securely taped to caddy, 2 cigarette lighters, spatula, 2 petri dishes of inoculum well identified with inoculum name, a rag. Mark caddy also with the strain of inoculum being used. Use sterilized cork bore to cut plugs of mycelium. Be sure to avoid making plugs of jell with no mycelium present. Avoid the center of the culture which may be contaminated with excretions from and dying cells of mycelium.

Tapers - need ½" masking tape and good attention span. Recorder needs list of trees by number to record trees which are inoculated, number of holes inoculated, and any errors made in the process. Date, location, orchard name, list of team members should be added.

 2. Plugger selects qualifying tree and four inoculation sites on main trunk. Avoid blighted areas, crotches, and green stripes which lay down the bark of the tree, known as Rhytidome initiators (about Rhytidome). Holes should be placed about 12"-18" apart. Wipe sites with alcohol rag. Sanitize cork borer and push and twist wrist a half turn til you hit hardwood. You can accumulate plugs for some time before emptying and flaming. If you can’t find four suitable sites, you may use just two, but most trees should be large enough to select 4 sites. Don’t get too far ahead of inoculators or the holes will dry out.

3. First inoculator uses less virulent strain (SG-1) of blight in two upper holes. Locate all four holes first. Fire both ends of the spatula, then slip one plug of jell onto first end and apply it to the top hole, mycelium in toward the cambium. Pat plug gently in. Wipe off end of spatula on pants or a cloth. Use other end to pick up second plug and apply as above in second hole from the top. If you put the mycelium in the wrong hole, mark the hole with permanent magic marker with the (SG-1) in several places around the hole and have the recorder make note of it. Go on to next tree. Fire your spatula for each tree. Keep petri dishes shaded so mycelium will not dry out and die. You may carry two dishes, but replenish supply from central source rather than trying to carry it all with you.

4. Second inoculator uses the same process using the more virulent strain (155) of blight on the two lower holes.

5. Tapers are the control people. Make sure the proper hole is inoculated with the proper mycelium. Immediately after your inoculator fills the hole, cover it with masking tape by circling the entire trunk or limb with the tape, doubling the tape over the hole. This keeps the mycelium from drying out. Make sure no trees that have been bored are missed. One taper follows the same inoculator all the way through. Quality control!

6. Recorder keeps record of all activity for the day.