Clonal propagation protocols of many species have already developed.
But through our experience propagation of cultivars of 5-10 and more year old trees and shrubs is very low effective because of a high contamination in vitro and low multiplication. So, dear collegues, does anybody have experience with commercial trees and shrubs cultivars or micropropagation? and if yes how is it possible?
Aloha Lidia. I have worked a bit with the Breadfruit Institute, who have developed a highly successful program of micro-propagation of breadfruit trees. Iʻm not the expert, but I believe that they material selection was highly important, selecting near the root/trunk interface. You might want to reach out the them and see if they have more information.
Let me know if you have trouble. Diane Ragone is my main contact there, but I donʻt know who was heading up their micro-propagation.
We do a similar method with poplar trees. We take dormant cuttings, root them in the lab (in soil) and keep the new plants in a greenhouse. Then, we collect stem cuttings from these plants, surface sterilize them with bleach, and put them into tissue culture. Contamination can be a real problem, you'll want to start with excess material.
Dear collegues, thank you very much for your answers! Amy thank you for your comment about cuttings. I've red about this way earlier and maybe I would try this.
One friend also advised me to put stems into the solution of cefatoxim or tofasolin (antibiotics) 300-500 mg\l for 3 hours and after that use usual sterilization procedure.
Tommorrow I gonna try it.
I think you use material that comes from the field. Order both to reduce contamination you can take cuttings of 20-30 cm with leaves (evergreen trees) and remove the leaves (deciduous trees). Introduction into mist chamber closed and await further breakouts. These new shoots have fewer micorganismos and easier sterilization
Carlos thank you for your answer.
You are right, we use field grown citrus trees for this work. My latest experiment with antibiotics was low effective. So I'm gonna try the way you offered.
yes, on the vine and the olive is applied. we have our experience. but, we are working with olive method "mist" two-three nodes
Long back, I did micropropagation of Manilkara hexandra using nodal explants from >25 years old tree...and we were successful in regenerating that. We took juvenile shoots coming out of the base of main trunk. Juvenile shoots did not pose much contamination problem. Its a fact that older the tissue problems are more. One can also try spraying a suitable fungicide well before collecting explants, this may reduce the inoculum load.
When the plant material in the field is heavily contaminated it has been effective to collect segments of branches and apply paraffin to the ends leaving them in a humid and warm environment to induce new shoots. These sprouts are cleaner and easier to decontaminate. In the presence of endogenous bacteria it is sometimes necessary to perform meristem culture to clean the explants.
macro throu micro propagation methods : Young branches or old branch cutting collected from selected tree and make vegetative propagation .there were producing new sprouts from cutting. those nodel segments used to ex plant. in various micro prorogation methods