Does anyone know why the same clone of plants gives different types of plants in vitro culture specially meristem extraction techniques. (eg'-single stemmed, multiple stems, thinned stems with clusters)?
Does anyone know how to get rid of contamination of in vitro plants ? fungal and bacterial contaminations
Success in multiplication phase depends on carbon sources concentration, light (presents, type and quality), mineral nutrition, permeability or fluidity of media, temperature, genotype, season, gas exchange, pH and specially plant growth regulator. Cytokinin chiefly BA has an important role in this phase. For your question, its better for you to set up every mentioned parameters in optimum condition for tested plant.
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Good luck
Reza Ghahremani
Dear Maduka Wehella you have asked two different questions. Since the second one is more "intriguing", I'll focus the attention on it. Several factors might influence the different responses for the same clone (considering that it's the case). The extraction of meristems is likely the most laborious and delicate activity in TC, performed in a microscopic level. Thus, the size of explants (meristems) are variable, bearing also a variable number of foliar primordia. Small variations in ontogenic age of meristems might also provoque variations. And don't discard those likely epigenetic variations associated to the explant source.
Dear Maduka Wehella,
Which plant species do you work on?
In some plant species, you need to set up a special micropropagation protocol for each clone. So, for the multiplication stage, an experiment regarding various influencing factors such as different concentrations of BA and GA is needed to achieve the desired result.
I hope this link is useful:
Article Indirect regeneration of Ficus carica by the TCL technique a...
Hi,
Ruhollah Abdolinejad its garlic, do I need to set up a special micropropagation protocol for each clone?
I am mainly looking to increase plant numbers using plant hormones. But I haven't succeeded yet.
Dear Maduka Wehella have you read Bhojwani's works with garlic? Regardless differences amongst genoypes might exist, and depending of your objectives, it would be worth to use a single protocol, likely not the best for each clone but a good one for the whole.
Again it depends on a particular plant species only. Generally, comparatively more BAP induces multiple shoots, and lower BAP elongates the shoots. GA we used for elongation of shoots, generally, it would not help in multiple shoot induction.
A good combination of BAP and the mild auxin generally help in the multiplication and elongation of shoots same time. It saves time, energy, and the cost of production.
yes however, I did trials with BA and NAA combination but I haven't got desirable multiplication rates@Mahipal Shekhawat
Maduka Wehella,
Agree with above answers, however it is not clear that how many shoots you have obtained.
For multiplication of shoots you can cut single nodes of in vitro shoots and transfer it to fresh medium with same or different PGR combination which in turn increase the number. In many species liquid culture also help in shoot multiplication.
As far as PGRs are concerned, yes BAP is one of the potent cytokinin but sometimes it need other auxins like IAA,IBA,NAA. You can also use Adenine sulphate along with BAP it might help.
All the best
Dear Maduka wehella
It depends on many factors like Carbon source, light etc apart from the plant growth regulator combinations. better to try with different types of cytokinins as well with different auxins at low concentrations
You can add filter sterilized coconut water ranging from 1 to 10 ml/L. I hope it will work.
Dear Maduka Wehella, as the first step for conducting a successful experiment, you better have a comprehensive literature review on garlic micropropagation. You need to test some introduced protocols, and then you can select the regeneration direction, including the best nutrient medium, explant, PGR combinations, and environment conditions (light, temperature). It might be a rather time-consuming procedure, but necessary.
The following links may be helpful:
https://journalbji.com/index.php/BJI/article/view/3036
Article High-frequency direct shoot organogenesis from garlic (Alliu...
Article Improved in vitro regeneration and propagation of Tunceli ga...
Best regards
I have used several multiplication ingredients and methods to improve the multiplication numbers of in vitro plants. Can anyone suggest any simple and effective method for me to try it out and or anyone already made any progress with the multiplication of in vitro plants with hormones ?
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