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Hello, I've been doing a lot of research and would love to get any feedback on my first attempt at a Mitragyna Speciosa tissue culture (which is legal plant/tree in my location). I know there are a few threads on here that have addressed this (shonman, Darklight), and I'm not sure if I should post on there and resurrect, or just start a new topic/thread; don't want to hijack shonman's thread, so I'm starting this as a new thread. I appreciate the time, energy, and resources that the members here have put into their practice, and would love to politely pick their brains; I understand that experience was hard-won, so I don't want to offend and ask for "info-freebies". My first hurdle is that I have limited mother plant material, and would really like to have at least one successful replication before I run out of plant. I've been working on this for a few months and have not had one yet; many were lost to contamination. A little about my method: -Working under plastic bin hood - WPM at label suggested strength - SIDNC (Milton Tablets) at 5000ppm for 15 minutes, but I wonder if my tissue samples are too large? Anyone tried 300-400ppm for 24 or so hours? -I also mix the SIDNC into the medium at 0.032%, but having a hard time adding it at the right time (ie. low enough temp to keep avail chlorine) without the medium ending up a little too soft. It's almost as if I need to heat/microwave the medium again, but that would destroy the SIDNC protection, right? For medium prep via microwave, pre SIDNC, how long do you let it boil? - 10% v/v coconut water - 4% Honey - 0.5 mg/l TDZ (this strength was at suggestion of fellow practitioner, open to suggestions on strength or using it at all?) Best way to dissolve TDZ powder? I've heard that petioles are best with this species (can anyone confirm that; goal is micropropagation) (I knocked off a leaf, so I decided to try that also, anyone had success with leaves of this plant?) What's the best way to position the sample on/in the medium? Size and cut of sample? Has anyone been able to micropropagate this with only two phases of medium (#1 medium=produce shoots AND roots, then #2 medium=hardening), OR does it need three phases of medium (#1=shoots, #2=roots, #3=hardening), OR four phases of medium (#1=callus, #2=shoots, #3=roots, #4=hardening) OR different combo...? Any thoughts on hormone use for these phases, I know 2,4 D is good for this particular plant to produce callus, but callus is not my end goal, Im not sure how to get callus to reorganize to shoots & roots. Air supply (hole in lid), I've read both nays and yays as to whether or not it needs it, right now, no holes, too worried about dust mites, etc getting in. I saw a thread post on here from a few years ago saying there might be an article/paper/protocol put out or published on this by Darklight and a few others, did that ever happen? I searched and searched, but could not find anything; I wish there were a manual for this;) After jumping in and doing a few trial runs, I can deeply appreciate how challenging and rewarding this can be. I wholeheartedly appreciate any and all feedback and tips! And I hope I've followed proper protocol for rules and etiquette on this post! I normally keep to myself, but after lack of success I realized I needed to reach out for help on this one. I only have pics of my most recent batch. The last one (#5) must be contaminated, can anyone "Name That Contam"?
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I have been experimenting with micropropagation for a while now...it is quite interesting!! Much thanks again to those who have lit the path before me, helping guide my steps... (...Darklight, Carol @ Kitchen Culture Kits, Shaman Australis Forum, and so, Torstein & friends...and the book, 'Plants from Test Tubes.) Dividing and multiplying the plants is very effective. They seem to put out roots, in some cases, in 1`.2 strength medium...oir, medium after the nutrients run out. They can be slowed down bv temperature, I have one batch that has not been transferred for five month!! (at 60F) Now, I am thinking about growing plants to a certain size, so they can be deflasked by others. The bigger the better up to a certain point. What might be the best way to do this part quickly? How big, is big enough? Here is what I am thinking: Divide explants until a certain number is reached. Root.,...in agar... Then, transfer to final sales container (any suggestions on these containers would be helpful). The medium in this final container in which the plant will be sold, would be full strength. They would have roots when placed in there. I would keep temperatures maybe at 70-75, and increase light. Theory being, that the plant would grow faster with light and full strength nutrients. Any thoughts on this are appreciated!! Thanks!
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Anyone have this one lying round? Specifically interested in Passifloraceae this week /applications/core/interface/imageproxy/imageproxy.php?img=&key=ed93ee4b8a158835e0af19ead9c794b11af03de360911f7858b9da338588c45d No, I won't be using the colchicine. Too toxic for safe use even here Plant Cell, Tissue and Organ Culture (PCTOC)December 2011, Volume 107, Issue 3, pp 451-459 Date: 01 Jul 2011 In vitro induction of autotetraploids from diploid yellow passion fruit mediated by colchicine and oryzalinM. M. Rêgo,E. R. Rêgo,C. H. Bruckner,F. L. Finger,W. C. Otoni
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I noticed the other day, that one of my Mitragyna Speciosa cuttings had grown roots from a leaf! Does any one know if it can generate plantlets in this manner, like Psychotrias , or African violets do? Any other suggestions for plants that can be propagated this way?
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This looks interesting.....and what a harvest, in such a short time! Seems to have a similar concentration as ordinary field cultivated ginseng root, it says.... The ginseng sides could perhaps be extracted and sold, as a standardized Ginseng root extract. Or, make and market your own soft drink/ energy drink/ ginseng tincture.... Wonder what other plants could be done like this? ............ Pilot-scale culture of adventitious roots of ginseng in a bioreactor system Sung Mee Choi, Sung Ho Son, Seung Rho Yun, Oh Woung Kwon, Jeong Hoon Seon, Kee Yoeup Paek Abstract A pilot-scale culture of multiple adventitious roots of ginseng was established using a balloon-type bubble bioreactor. Adventitious roots (2 cm) induced from callus were cultured in plastic Petri dishes having 20 ml of solid Schenk and Hildebrandt (1972) medium containing 3% sucrose, 0.15% gelrite, and 24.6 μM indole-3-butric acid. An average of 29 secondary multiple adventitious roots were produced after 4 weeks of culture. These secondary roots were elongated on the same medium, reaching a length of 5 cm after 6 weeks of culture. A time course study revealed that maximum yields in 5-l and 20-l bioreactors were approximately 500 g and 2.2 kg at day 42 with 60 g and 240 g inoculations, respectively. Cutting twice during the culture increased the total amount of biomass produced. The root biomass in a 20-l balloon-type bubble bioreactor was 2.8 kg at harvest with 240 g of inoculum after 8 weeks of culture. The total saponin content obtained from small-scale and pilot-scale balloon type bubble bioreactors was around 1% based on dry weight. Inoculation of 500 g fresh weight of multiple adventitious roots into a 500 l balloon-type bubble bioreactor with cutting at 4 and 6 weeks after inoculation produced approximately 74.8 kg of multiple roots. The ginsengnoside profiles of these multiple adventitious roots were similar to profiles of field-grown ginseng roots when analyzed by HPLC.
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On the podium are five sterile culture containers of Acacia acuminata. Tubes to be auctioned separately. These are aseptically germinated seed, and are suitable for further sterile work, or for deflasking into a suitable environment for garden growth. Once you receive them they are yours to do with as you wish Funds from this auction will be donated to the Seed Saver's Network, to continue their work teaching and mentoring communities in Australia and overseas, encouraging groups to maintain their local, biodiverse and sustainable food resources. Jude and Michel do some of the best outreach work I have ever seen and they have inspired me for several decades now http://www.seedsavers.net/ Auction notes: if you are a successful bidder you accept the terms and recommendations below Plants are approx 2-4cm tall with two explants per jar. They will have another few weeks of growth in them under fresh coolwhite fluro tubes, or indirect light about 200-250 µmol photons/m2/second ( ie about 10-20% of daylight strength- more could cook them ). If plantlets get taller than 7-8cm it can become difficult to remove and handle them aseptically.Auction open to addresses within Australia only, with the exception of WA. I will not ship to WA for quarantine reasons. Cultures are functionally sterile at the time of shipping, but media contains PPMCulture tubes are 150ml, with 50ml culture media containing 10g/L gelcarin. The high gelling agent amount will help, but not entirely prevent root and stem damage during shippingI won't be held accountable for their success in further culture as that relies on factors outside my control. Simple aseptic teks have previously worked for minimal replication of this species. If you post your tek and pics to SAB myself or someone else may be able to help, but that depends on time available to me and clarity of your requestsAuction bid includes shipping costAuction bids open at $20 per jar, and auction closes at midnight on Sunday 28th April 2013 Tube #1 Starting bid $20 Tube #2 Starting bid $20 Tube #3 Starting bid $20 Tube #4 Starting bid $20 Tube #5 Starting bid $20