These cannabis plants were healthy and green for 8 weeks and they started browning when I put them in rooting media. No rooting achieved and they are turning brown.
Hello Poorva,
There are multiple factors that might lead to this. assuming that your light, temp, humidity did not change i have observed that some plants react poorly when subjected to high auxin concentrations. IBA generally causes such browning, you might want to decrease the concentration or give a pulse treatment.
Dear Poorva Vyas it does mean that you have used your protocol and it functions quite well? or at least in a different way. Isn't it?
Other question, is it possible that cultures are contaminated? In both pictures look like if microorganisms were growing in culture media.
High concentrations of auxin can be a possible reason. Moreover, As you sub-cultured the explants in a new media continuous detachment of explants from culture medium during sub-culture under laminar where the temperature is higher and relative humidity is lower than in-jar atmosphere can be an assumption just according to experience!
Dear Poorva Vyas , in rooting media, it needs an only a small amount of auxin only. The best is IBA. No need to put any kind of cytokinin. however, maintain the MS medium in full strength. also, I noticed that you haven't separated your plants nicely before put it into the rooting media. separated individual shoots and placed shoots in new media with a little bit of distance. hope then it works.
Hello Poorva Vyas Poorva Vyas, I was also facing the same problem with watermelon, then I changed the concentration of sugar in my media (I decreased from 3% to 1%), then hopefully it worked. you can try to decrease the sugar concentration. then I changed the cytokinin/auxin type and ratio. you can try IAA or IBA, and try to make an optimization experiment with a big distance.
I hope these will be helpful for your experiments.
Poorva Vyas
Unfortunately, they are dying and dead. The medium has problem. You should follow an established protocol in a paper to do it. It is hard to give suggestions here, because different plant species has its own optimum medium formula. I checked online, there are different Cannobis species. ex. C. sativa and C. indica (1). What is your Cannobis species? There is one paper about C. sativa tissue culturing. See below link (2).
(1) https://en.wikipedia.org/wiki/Cannabis_sativa
(2) Article Tissue Culture of Cannabis sativa L. and in vitro Biotransfo...
There may be many reasons behind this but first and foremost as said by Ricardo Julian Licea-Moreno sir, I have also observed contamination in the culture media due to which the plants may turned brown. You can also use earlier established protocol as suggested by Yuan-Yeu Yau sir.
I have some other points to mention-
1) You have not mentioned the concentration and strength of MS media as many a times reduced strength i.e half or quarter is reported to be beneficial.
2) Sucrose at 1% concentration is mostly suitable for rooting. Higher concentrations adversly affects rooting.
3) You can try liquid medium instead of agar gelled which causes contamination and slow rooting.
4) Although IBA is wildly used for rooting, many plants induce better rooting in presence of NAA.
5) If the growth of plant is good then you can take shoots after 6weeks no need to wait upto 8weeks.
6) If Browning is still appeared you can give charcoal or PVP treatment which prevents any leaching out of any type of exudates.
7) And lastly you can try rooting using natural planting substrates like cocopeat, vermiculite, sand, soil etc. Which is being used nowadays as it will decrease the micropropagation time.
All the best.
There are many factors which influence on the rhizogenesis, but for browning of culture is most probably due to phenolic secretion of the tissue used as explant . therefore you can use activated charcoal to control the phenolic secretion thereby rooting also may takes place.
best of luck
Hi, probably your media contaminated. Plz give high resolution picture and it will be easy to conclude.
I think the rooting media after the inoculation elongated plant, it will be produced phenolic like compound so you have to add some additives (arrest in those likely compound secretion) in the rooting media.
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