HERBACIOUS PLANT GRAFTING MANUAL
Germinate pea seeds
- Scarify seeds in concentrated sulfuric acid for 5 minutes.
This is done to soften the seed coat. If omitted, seeds may take longer to
germinate and germination may be less regular
- Imbibe seeds in aerated water for 24 hrs.
Same as above
- Plant seeds in wet vermiculite 1-2 cm deep.
If seeds are only pushed into the surface, roots may not grow well,
especially if air is dry (in this respect the picture on the right is
- Leave to germinate in vermiculite dark until shoots are 2-3 cm long.
Too short seedlings are hard to graft, extensive etiolation is not desirable
because we want the plants to be as close to "normal" as possible. Using
other medium than vermiculite is probably OK, it is convenient to use something
that does not stick to the roots too much
- Prepare pieces of tubing that will fit tightly the epicotyl,
(PE is best, possibly other materials) make them 1 cm long and
slit-cut them along their axis.
Tight fit is important, because this is the part that will hold the
grafted parts together. If they are not held tight, air gets in, dries the
area of contact and the scion dies. Use several diameters, if necessary
- Isolate two seedlings that are to be grafted and cut them with a sharp
razor in the middle of the epicotyl at an angle of 20-30 degrees.
Blunt blade may cause problems in healing, angle increases the area of
contact and makes attachment easier
- Attach a piece of tubing to the stock, so that half of it is empty, ready
to accept the scion
This is a moment to check how well the tubing fits. Choose a size that will
have to open slightly to accomodate the stock
- Slide the scion into the tubing
Make the contact with the stock as tight as possible. Don't push too hard,
so as to damage the tissues
- Place grafted plants back into vermiculte
Wait for the grafts to take
- Leave the grafted plants in a moist environment with moderate light
This can be done by covering them and keeping the vermiculite wet.
You may increase light to normal levels after a few days.
- Buds in the axils of the cotyledons will start breaking, cut them off
If these lateral buds are left to grow, they overtake the whole plant
and the scion may die. Their growth can also be prevented by cutting off
the cotyledons, but we want to keep them there for a few days to keep the
- After 7-10 days it should be clear which grafts have taken. Cut off
the cotyledons and use those grafted plants as desired
In our experiments we transferred them to hydroponics
About this manual
This manual was prepared by Matej Lexa. I would like to hear from you in case you find it useful in any way.
If you find this interesting enough to use in class, go ahead !
In case you need a reference for a publication, use:
Matej Lexa, John M. Cheeseman
Growth and nitrogen relations in reciprocal grafts of wild-type and nitrate reductase deficient mutants of pea (Pisum sativum L. var. Juneau)
Journal of Experimental Botany 48, 1241-1250 (1997)
This document was created on April 29, 1996.