As a UG student I was able to culture Duranta gold (A common Fluorescent green hedge plant). I also tried out with Ficus guavajava (Guava tree), but was not successful due to the high phenol content.
How different is it for aquatic plants? Could you forward me some literature on it.
Starting out
1. I have some 100 ml sample containers (Tar sons) which I use for storing water samples. They may be resistant possibly till 70 or 80 and are pretty cheap. Don't know it they will withstand Autoclave treatments.
http://www.tarsons.in/?a=c&s=510010. Will also bring them during the next ASK meet.
2. Dimethyl sulfoxide and Agarose can be sourced out from chemical stores like Vasa Scientific (Malleshwaram / S.P. Road)
3. Scalpel, Forcep, Surgical gloves can also be sourced from Vasa
4. Multi-Vitamin tablets and antibiotics must be the more expensive part. We can order them based on the needs.
Sandeep - Crypts, one or two types actually ! Might try Narrow leaf Java Fern also, but that needs a liquid medium.
You need emersed specimens for tissue culture, submerged specimens will simply not do.
Too bad I replanted all my emersed Crypts just a few days ago, didn't know I'd end up with a few days spare time.
Ravi - There's hardly any literature worth the while about aquatic plants. Thanks for the tip about agarose and Dimethyl sulfoxide.
What was the biocide you added to the media ? Mercuric chloride is one but that's toxic, I'd rather take my chances without any biocide.
I feel we have to go with glass bottles unless someone find us a good source for microwave safe plastic bottles and lids.
Any source for glass bottles ?
If anyone has emersed grown plants that could also be tried.
Which is better Isopropyl alcohol or Ethanol for sterilising ?
Madan Ji...normally for culturing,cotton rolled tight , covered with aluminium foil is used to cap the cultures.
Can this be used? Metal caps will leave the bottles airtight not letting the cultures to breathe.
Madan Ji,ethanol is preferably better as I learnt in my little experience in culturing.
Also, rightly said by you, mercuric chloride will be toxic, better to start without biocide, instead Sodium or calcium hypochlorite can be tried, but we need to check more on them before using.
Auxins can be added for the explant growth and regulation, I believe it should be easily available.
But the most difficult step would be storing the cultures as they will need a complete aseptic place to grow atleast for a month,or the enriched media will attract a lot of foreign bodies
Last edited by Cherry on Wed Jul 25, 2012 4:26 pm; edited 1 time in total
Madan, As Nayak pointed out, Vasa Scientific in Malleshwaram is very much accessible to me. Will check for the prices of both Agarose and Dimethylsulfoxide.
We used Kanamycin, which is detrimental to most bacteria (main contaminants). I will also check for other alternatives from my friends.
We added commercial macro and micro nutrients (Not Azoo) to agar (similar to dosing of ferts), alongside with Auxin and Cytokinin.
Shanai, As you suggested, the biggest problem will be the storage and aseptic condition. We can always fume dedicated boxes (few small ones) with formalin and store it. Wont it work?
The most effective agent for cleaning the explants would be to rinse them with Methanol/ dilute Hg2cl2 for a minute or so.
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