Hello Mehmet, I am working in culture in vitro of cotton. I do subculturing every 3 o 4 weeks when they are callus or shoots. But when the explant starts to growth in vitro the subculturing is every week for 3 weeks, thus avoid the oxidative browning. But this is depend if you use antioxidants.

I hope that I have answer your question.

We are working to propagate some citrus rootstock in vitro. We noticed that subculturing every 4 weeks is better in growing plants because the medium of cultivation during this period is depleted. However, the period of subculture depeded on the plant species and culture conditions.

It depends of the stage in which are you working, the species, the system that are you using as well as the health of plant materials, amongst the most important. The length of subculture is normally higher during in vitro establishment than in proliferation. In semi-solid media is also usually higher than with liquid media. In temporary immersion you can enlarge more the subculture than with static systems. While, when decay symptoms begin to appear it normally means than subculture is neccessary.

Thanks everyone for your answers Guido Pillaca Siham A. Salim Ricardo Julian Licea-Moreno

I understand from your answers that there is no specific time to subculture, and that determining the subculture time and repetition, together with the variables you count, is observational or depends on the content of our study.

That the dosage and combinations of plant growth regulators I observed in my reproduction study of some endangered species of lilium were very effective at the time of subculturing. I think the time to subculture (browning, yellowing, stopping growth, etc.) due to the bulb scale of my explant source was up to 8 or 12 weeks.

Dear @Mehmet I have to recognize that 12 weeks seem to much for multiplication experiments. It seems more proper for conservation. Anyway, remember that you're who decide what is the best for you and your cultures. However, you must always to consider costs (not only money)/ benefits.

Good luck !!!

If your objective is achieving a maximum plantlet yield, then whatever method reaches your goal is OK. Of course you must keep tabs on costs and yield quality.

Good luck!

Variable: 1- Depends upon goal 2- Varietal response 3- Medium with hormonal combination 4- Protocol of micropropagation steps -a- Multiplication followed by elongation and finally rooting, b- Multiplication and elongation simultaneous followed by rooting.

Of course, it is the browning by photooxidation followed by the aseptic status of the cultures

It depends on how long you want it to be in the container...supplements of activated charcoal are effectively keep the cultures from long period as it adsorb the waste substances and phenolic compounds from plants....also you can increase the gel to reduce the availability of water and nutrient to the plant so that they last longer in in vitro system....

I also add plant preservative mixture, PPM to avoid contamination...

It is better you get yourself breathable cap which has filter with it...please PM me for more if you want to know as it is prohibited for me to tell you the details in researchgate