sativ Posted April 13, 2010 Report Share Posted April 13, 2010 (edited) Hello ;) I'm working with TC for quite long time and i want to share some photos with you. There are also many misinformation about tissue culturing- usually opinios are made by people what never did anything in tissue culturing subject. I want to make some of "dark side of TC" brighter ;) If you have any question about medias/sterylisation/other infos- i will answer them, but some issues are my "patent" and i will not share them, but there are only some most special things, not interesting so much for amatour. and here- some photos ;) : How it staretd about 2 years ago...: Now little bit more plants: First- classic ;) VFT from single seed typed from more of them due to nice colour and shape: [now i have little bit more of them- ~300 plants] Next classical plants- D.regia...hard to multiply... some d.adelae d.pulchella "scott river" likes to flower ;) more VFT... Something more interesting- N.bokorensis.. gymanphora.. madagascarensis... truncata... tiny bongso... ramspina... H.minor... In this moment i have ~80 different species in TC. If you have anything interesting in tc i can spare for my TC plants. Now i'm looking for as many nepenthes and haliamphoras seeds as it is possible- if you have any, fresh! write me ;) I hope that you liked this small part of photos i have- i will try to update this topic sometimes. Regards Edited April 13, 2010 by sativ Quote Link to comment Share on other sites More sharing options...
dudo klasovity Posted April 13, 2010 Report Share Posted April 13, 2010 Wow that is so cool! Very nice developed plants! I mostly specialize in drosera genus, just starting to do some Nepenthes on different media. I started with TC about a year ago. Can you tell me how fast do the nepenthes grow in TC from seed? Drosera grows very quickly, but never had a chance to compare nepenthes growth. For example, these seedlings of nepenthes muluensis are about month old. I think it is an ultrahighlander, so I give it colder nights so if the growth is too slow I guess it is due to media (I am testing my own composition) and maybe the chemicals need to be altered..?. Here is the pic BTW, I will have the requested swap TC species ready for you by the end of this month:-) Quote Link to comment Share on other sites More sharing options...
sativ Posted April 13, 2010 Author Report Share Posted April 13, 2010 Start of neps from seeds is very slow. They need some time [3-4 months] to grow to 2-3 cm, than it start to go faster, due to new medium what can be used. Tiny seedlings are very delicate, they can die in 1 night from unknown reason.... My newest batches of seeds are germinating in the less concentrated medium than before, but too big dillution is also not good due to osmotic stress...I don't think that colder nights are really needed in TC- temperature impacts dark phase of photosynthesis in plants in-viivo, but in vitro plants don't need to photosynthetize much ;) Best medium for neps is very hard task, each specie has a little different preferences... But they grow well on every medium with low concentration of NH4+ [30% MS range] Did you transplanted your seeds in cothylion stage? It is risky i don't do that until they have 2-3 leaves. Medium what you use is WPM based with aminoacids? So much tiny plantlet's cannot tell much, just curiuos shot ;) But they lokk quite well. First shot of N.bongso what i made: Quote Link to comment Share on other sites More sharing options...
dudo klasovity Posted April 13, 2010 Report Share Posted April 13, 2010 (edited) Thanx for reply:-) So it takes some time for them to develop and take off. Yes, I did re-plate them in cothyledon stage, because some of them started dying and I needed to change the medium. Old medium had 1ml PPM in it, now i dont use PPM. I have read that TC plants dont photosyntetize (much), which is understandable, since there is 99% no air exchange and they do not need carbon from aerial CO2 when they get it from sugar (and other organic compounds). Nevertheless, I thought the night cold temperatures that some plants require are not only good for dark-phase of photosynthesis, but also for slowing down metabolism, thus getting 'night rest/sleep' needed to recover. So what you say if i keep an ultrahighlander nepenthes, grown in TC at 23C at all times, it does not know the difference? Interesting... For neps I try to use medium: Macroelements: 230.0mg Ca(NO3)2.4H2O, 300.0mg (NH4)2SO4, 150.0mg KH2PO4, 80.5mg MgSO4, 40.0mg CaCl2 Microelements: 5.53μg CoCl2.2H2O, 15.0mg FeEDDHA, 5.53μg CuSO4.5H2O, 1.4mg H3BO3, 0.18mg KI, 3.86mg MnCl2.2H2O, 0.055mg Na2MoO4.2H2O, 0.9mg ZnCl2 , 3.73mg Na2SO4 Vitamins: 1.0mg thiamine hydrochloride, 1.0mg riboflavin, 0.66mg pyridoxine hydrochloride, 0.66mg niacin, 1.6mg Ca-panthotenate, 66.6mg myo-inositol pH=5.62 I might need to mix another, but first I wait for the behaviour of this species:-) I mix this one assuming they dont like high concentration of NH4NO3 that is in MS, even diluted (wrong ratio to other compounds), so I use different forms of NO3 and NH4+ in lower concentrations.Please, take this for one of many experiments. Edited April 13, 2010 by dudo klasovity Quote Link to comment Share on other sites More sharing options...
sativ Posted April 13, 2010 Author Report Share Posted April 13, 2010 Your macro's looks very similar to Knudson C 1946 medium in ~30% concentration. That's too low concentration- try to doubt the macros, microelements taken from MS- well, concentration is also good. Rich blend of vitamins ;) Medium looks very well, but try to use stronger ;) I use PPM for nepenthes and i don't observe reduced growth rate. I made one time some medias without PPM- and that was one time when i get perfect airborne contamination what killed ~100 small plantlets of nep's ;) So now i don't use PPM only if it is necessary- like some drosera leaves. I like PPM very much, wirking with it is really easier, even with laminar flow hood... About ultrahighlanders in TC- just send me explant of N.villosa, macrophylla- i will try to answer do they can grow in 20*C all time ;) I don't think that plants in TC have to recover or slow down- they don't have to do many mathabolic work, becouse they have in their tissue ready elements for growth. Quote Link to comment Share on other sites More sharing options...
Stefan P Posted April 13, 2010 Report Share Posted April 13, 2010 Amazing - if TC increases the circulations of plants that are otherwise very rare then it cannot be a bad thing surely Quote Link to comment Share on other sites More sharing options...
dudo klasovity Posted April 13, 2010 Report Share Posted April 13, 2010 sativ: thanx for advice, I will think of something else then, as far as macros are involved. PPM used to help a lot back when I had not have elaborated process of in-box work and subsequent steps. When I get contaminations, I will consider to use it, meanwhile it will be sitting in the fridge. It is nice to know that nepenthes are less sensitive than drosera or pings. Sorry to hear about your past loss of plants, but we both know this is inevitable sometimes in the learning process:-) As for N. villosa and N.macrophylla in TC giveway---hahaha, you are a funny guy:-) (Not ruling out that when I have it, I will send some:-) I will really try to grow some ultrahighlanders in steady temps of 23-24C and if they thrive I will believe in 'the absence of common requirements' in TC :-) Quote Link to comment Share on other sites More sharing options...
sativ Posted April 13, 2010 Author Report Share Posted April 13, 2010 There are some crazy guys what can get N.villosa and macrophylla seeds, i think that was the way from where we have them from BE and wistuba in TC....Introduction from plants is very hard but possible [N.bokorensis was introduced from my plant] Problem with rarest neps and TC for amatour is money... They are to valuable to cut them in parts and soak in hypochlorite ;) My tests ate most of my collection what is now "rising from ashes", but i really want to try with them too ;) Do you find PPM toxic for droseras and pings? I also use it for them and they grow like crazy... And do you use PPM oryginal from USA, or cheaper from other distributor? Quote Link to comment Share on other sites More sharing options...
dudo klasovity Posted April 13, 2010 Report Share Posted April 13, 2010 (edited) I guess you can get many species already in TC from commercial trade but I would like to learn the whole process of introducing different neps species into TC from seed (tissue is much harder, of course-need for antibiotics and other chemicals, hypochlorite can be too harsh for them, maybe the use of colloid silver in combination with benzalkonium chloride and some ATB can be more selective and effective)... Neps from tissue I am only intending to try once I get sterile culture from seeds. I have my PPM (along with hormones) from Carol Stiff from USA (so I guess-I hope, they passed the quality control) :-) I do not think it is exactly toxic for drosera and pings , but my comparison experiments I did with non-PPM media showed that plants grow even more crazy without it. Of course, there is a compromise needed when contamination gets into play (which is not my case right now-knock on the wood;-)) Edited April 13, 2010 by dudo klasovity Quote Link to comment Share on other sites More sharing options...
sativ Posted June 4, 2010 Author Report Share Posted June 4, 2010 I'm really glad from my new nepenthes aclymatisation medium ;) N.madagascarensis passaged 9.05 Quote Link to comment Share on other sites More sharing options...
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