Tissue culture consultant Cali?

Can anyone recommend a tissue culture consultant in Northern California?

I am looking to set up a small legit lab with the capability of storing a few thousand cuttings for after harvest selection and long term storage of mothers and fathers. Most of the strains are pure sativas and this seems like the most logical way to keep so many plants for so many months/years with the least amount of work/energy/Space.

I posted a request for a consultant on zintro dot com yesterday. Had 20 consultants from around the world respond lol, going to see where that goes, these guys seem SUPER competitive with each other.

I know a few with TC experience, but not on Cannabis.

Just to caution you, it seems like you might be thinking that TC would require less total labor/infrastructure/cost resource usage than perpetual mother/clone rotation. This is debatable, since TC involves a large infrastructure setup and expensive (sterile equipment) ongoing supplies. I dont think you realize the constant amount of transferring/subdividing culture bodies that is necessary to keep them alive, unless you can work out a reliable cryopreservation protocol (which I'm not familiar with so I can't comment on feasibility).

While TC would reduce some space needed, avoid plant count concerns, and eliminate most whole-plant pest/pathogen concerns... TC also carries with it increased labor and infrastructure costs as well as increased susceptibility to normally nonpathogenic bacteria and fungi.

The safest and most reliable method of germplasm preservation continues to be seed banking.

I appreciate your response

I have allocated $8,000-$15,000 for the project. I am not thinking it will be "cheaper" to set up and I have no problem with investing in infrastructure.

It will actually be WAY less labor intensive. I could understand your point if I was going to be keeping thousands of plantlets alive versus so many cuttings alive, but that is not an accurate comparison. I will take the plantlet off of each plant just before harvesting. I'm sure they can "rest" in the first stage of TC for a few months allowing me plenty of time to narrow 1000 plantlets down to 50 winners before I actually have to do any "work" or transferring them to the next level.

Also this is a long term plan because I will only add more greenhouses, I'm thinking 4. Two dedicated to males and 2 for females. All of the chosen females will then be tested outdoors and grown 200 gallon (4') pot to test outdoors after the greenhouse testing.

I am not concerned with plant counts.

Fair enough, just raising some associated concerns. I would expect to need to subculture a callus mass or plantlet at least every month, since the plant hormones in the media will degrade over time, it might not work to stretch that any longer. Cool temps and low/no light (for plantlet/callus) could be helpful for holding them as long as possible.

Seems like it should work. But I'm curious, for the purpose you describe, what do you envision being the benefits of going with TC over low-temp cutting-sized plant maintenance or 2-3 rounds of cutting cycling?

If all I had was 1 month I could get it done.

1.5 week dry
2 week cure
.5 week sent off to be terpinoid/cannabinoid tested

before I posted this thread I was going to actually post a question asking about both TC and low temp cutting setup. Could you point me to some good info on this? I looked into it but didn't find enough info to feel I could for sure do it. Another thing I worry about is I am 100% off the grid. I could set up a propane fridge but not an electric at the moment.

The main reason for not wanting to do multiple rounds of cuttings is honestly I think I would just "drown" in that additional work load and stress of managing multiple thousands of cuttings for half the year. I will already be managing a 7000 square foot outdoor farm. Most of this work will be going on When I will be managing that harvest from September-January. I just really don't want to worry or stress about these things because if it takes too much time from my farm work then it REALLY starts costing me $$, this greenhouse stuff is not for $$.

Tissue Culture Technicians get paid a lot of money somewhere upwards of $80K + a year. That isn't including the supplies and facility needed to manage something like what you are talking about.

As for the refridge freezer check out SunFrost. They build solar powered refers/freezers to your size specification or application. They are awesome, all the people in the hills up here oogle and aww all over them.

Well the whole point of paying a consultant is for them to help me setup a design a workable method. I have PHDs with 30 years experience in Malaysia and India telling me they can set this up cheap and efficient lol, I'm taking my time With them. The idea would be to have a small lab where I am the lab technician.

Thank you VERY MUCH for sunfrost info I will DEF check that out!

Okay, I understand a little better. If you are able to have someone run a TC operation in the course of 1-2 months for the budget you stated, then that could be an option. I suspect that in that short a time frame, the process would be basically taking cuttings and rooting them with possibly also making a backup callus bank (if freezing and revitalizing is viable, this might be the best bet, as you would get both short-term and long-term backup). The thing about TC is that it's all about getting the hormone balances right and sometimes all sorts of other fiddly factors for the individual species and even potentially down to the individual genotype, so proving the reliable effectiveness of protocols for callus formation, root generation, and shoot generation are all necessary before I'd recommend it for the type of undertaking you describe. This means probably at least 6 months development time as well as employing 1-2 full time people at 40-70k per year each. So unfortunately we're back to what you already know, which is that you need someone with the right (meaning Cannabis-specific and reliable) experience for the TC approach which I'm sadly not able to point you toward. I'd be hesitant to trust easy promises in this regard, worst case would be more than a year just optimizing protocols to be able to count on TC as even a short-term preservation technique. Even if you are able to receive setup and training and act as the lab tech yourself, the actual implementation might be as much work or more than just watering and pruning cuttings or dwarf plants.

I've not been able to directly test as of yet, but research appears to indicate that multiple factors could be utilized to inhibit growth and therefore make a set of 'cuttings' grow more slowly and be easier to manage. This is not usually what is desired, so obviously the research is usually coming at this from the opposite angle, and the growth rate will need to be managed against the minimal health of the plants, since you want to slow them as much as possible but not kill them.

Cold temperatures, in addition to (but apparently not directly because of) drought stress inhibit photosynthesis through various mechanisms, and the net result is significantly decreased growth rates. A sea (or better yet, a sloping wall) of small plants kept at cool temperatures (especially at night) might have their growth reduced by more than 50%. The plants will be transpiring much less, so they will be using less water also. I would recommending fully rooting the cuttings and planting them in small containers, once the roots develop they will effectively be experiencing drought stress because of becoming root-bound. It looks to be mixed as to whether dark cold periods or high light are more growth inhibiting, but since you are likely to be limited in power generation and might need to prioritize cooling over light, that choice sounds like it has already been made for you by the circumstances. If the 'cutting' death rate is unacceptable (because they aren't tolerating the conditions), the addition of a growth inhibiting PGR like paclobutrazol could help decrease their sensitivity to the cold and dry while keeping the growth rate low and internodes short. Salicylic acid may also have photosynthesis-inhibiting effects.

Great recommendation on the SunFrost, that could be something to look into. However you might be able to get away with evaporative cooling, which uses very little energy and is greenhouse-friendly. A good target to shoot for would be getting near or below 50F (10C) during the night (ideally longer than 6h), hopefully while also keeping day temperatures from getting too high, below 70-75F (21-24C). A circulating (water) chiller could also be an option for cooling the plants, particularly at the root zone.

Below are some links to related papers, although unfortunately not all of them are freely available, and this is not an exhaustive search.

Chaves et al (2002) How Plants Cope with Water Stress in the Field. Photosynthesis and Growth. Annals of Botany 89: 907-916

Martin, Ort, and Boyer (1981) Impairment of Photosynthesis by Chilling-Temperatures in Tomato. Plant Physiology 68: 329-334

Flexas et al (1999) Analysis of the Relative Increase in Photosynthetic O2 Uptake When Photosynthesis in Grapevine Leaves Is Inhibited following Low Night Temperatures and/or Water Stress. Plant Physiology 121: 675-684

Allen and Ort (2001) Impacts of chilling temperatures on photosynthesis in warm-climate plants. Trends in Plant Science 6: 36-42

Martino-Catt and Ort (1992) Low temperature interrupts circadian regulation of transcriptional activity in chilling-sensitive plants. PNAS 89: 3731-3735

Kee, Martin, and Ort (1986) The effects of chilling in the dark and in the light on photosynthesis of tomato: electron transfer reactions. Photosynthesis Research 8: 44-51

Ortiz-Lopez et al (1990) The involvement of the photoinhibition of photosystem II and impaired membrane energization in the reduced quantum yield of carbon assimilation in chilled maize. Planta 181: 78-84

Huner, Oquist, and Sarhan (1998) Energy balance and acclimation to light and cold. Trends in Plant Science 3: 224-230

Janda et al (1999) Hydroponic treatment with salicylic acid decreases the effects of chilling injury in maize (Zea mays L.) plants. Planta 208: 175-180

Berova and Zlatev (2000) Physiological response and yield of paclobutrazol treated tomato plants (Lycopersicon esculentum Mill.). Plant Growth Regulation 30: 117-123

Wang (1985) Modification of chilling susceptibility in seedlings of cucumber and zucchini squash by the bioregulator paclobutrazol (PP333). Scientia Horticulturae 26: 293-298

MountZionCollec said:

I have PHDs with 30 years experience in Malaysia and India telling me they can set this up cheap and efficient lol

Click to expand...

If you do hire somebody remote make sure you only pay a "fixed price" and them getting paid is contingent on you being happy and if they can deliver on what they say they can.

Otherwise it will be a fiasco.

I know upwork.com has the ability for "fixed price" jobs and I use them a lot. Not sure if Zintro has that option? I would not pay more than 3k for a project where they are working remote in India and it is crystal clear to them that no results=no payment. That would keep them honest and motivated. Malaysians/Japanese will cost 5x more than Indians or Chinese freelancers/moonlighters. These remote worker websites are known to have people promising the world and delivering nothing, all the while billing you weekly. Good luck!

This may sound dick-ish, but there is no way anyone with actual cannabis TC experience is going to provide protocols and training, even offsite, for 3K. Pay after work contingent on your happiness? Ya right - way to many places for you to fuck things up along the way, so why would anyone provide you all that info and trade secrets when payment is contingent upon you being successful? For me personally, sharing the years of tinkering and learning in lab at my expense without payment upfront would be a non-starter, I simply wouldn't do it.

Tons of people will tell you it's easy and doable, and all of them have zero experience with cannabis TC. This is not so cut and dry with public protocols worked out and proven solid like other crops, not to mention that some cultivars take to it well and some don't.

It's way more complicated and resource intensive than you are anticipating, and long term storage of callus with regeneration is not easy or practical, despite a couple of papers on the topic claiming so. Different genotypes behave differently, and very few re-shoot easily- which is why the papers on the subject use a single cultivar.

What you require guarantees that every clone can be pulled out of stasis, or back into a full plant or there is no point. You are better of storing cuttings in small well labelled pots in a library rather than TC you will have better results and won't waste a bunch of money.

Hope that helps,
-Chimera

Chimera said:

This may sound dick-ish, but there is no way anyone with actual cannabis TC experience is going to provide protocols and training, even offsite, for 3K. Pay after work contingent on your happiness? Ya right - way to many places for you to fuck things up along the way, so why would anyone provide you all that info and trade secrets when payment is contingent upon you being successful? For me personally, sharing the years of tinkering and learning in lab at my expense without payment upfront would be a non-starter, I simply wouldn't do it.

Tons of people will tell you it's easy and doable, and all of them have zero experience with cannabis TC. This is not so cut and dry with public protocols worked out and proven solid like other crops, not to mention that some cultivars take to it well and some don't.

It's way more complicated and resource intensive than you are anticipating, and long term storage of callus with regeneration is not easy or practical, despite a couple of papers on the topic claiming so. Different genotypes behave differently, and very few re-shoot easily- which is why the papers on the subject use a single cultivar.

What you require guarantees that every clone can be pulled out of stasis, or back into a full plant or there is no point. You are better of storing cuttings in small well labelled pots in a library rather than TC you will have better results and won't waste a bunch of money.

Hope that helps,
-Chimera

Click to expand...

Thanks a lot for the comments chimera it does help. You are absolutely right the whole idea hinges on being able to pull any plant I want back out of stasis and if that is no where near guaranteed then TC would not be good for what I'm trying to do.



Would you be able to comment on the idea of rooting in the fridge or placing the cuttings in the fridge to slow down the process Or would you still recommend small pots in a proper library?

Thanks again

A buried refrigerated storage container lined with rigid foam insulation, to store clones after (not during) propagation.

A bank of florescent lights for the above.

You can make a deal on decent lab work after smoke tests skip tlc.

2 honda eu2000's and gas to run it all for the next 10 years.

you can do that with 15k, prolly the best option no matter how fat pockets are..

TomHill said:

A buried refrigerated storage container lined with rigid foam insulation, to store clones after (not during) propagation.

A bank of florescent lights for the above.

You can make a deal on decent lab work after smoke tests skip tlc.

2 honda eu2000's and gas to run it all for the next 10 years.

you can do that with 15k, prolly the best option no matter how fat pockets are..

Click to expand...

Thanks Tom, really amazing information thanks for sharing.

You say for storage after propagation but not during, so you take cuttings in flowering for storage this way or did I misunderstand? I've read 45-50 degrees is the temp to shoot for, would you agree? I've read you can root in these conditions, do you root before or just stick the cuttings the medium when placed in Container?

Well my pockets are not fat lol, but enough so after harvest this year that I'm able/willing to invest in my young farm. I misunderstood TC, and appreciate everyone for sharing there information so I didn't make a bad investment!

http://home.olemiss.edu/~suman/Geneticstability.pdf


Pak. J. Bot., 41(2): 603-608, 2009. A MICROPROPAGATION SYSTEM FOR CLONING OF HEMP
(CANNABIS SATIVA L.) BY SHOOT TIP CULTURE
REN WANG1*, LI-SI HE1, BING XIA1, JIN-FENG TONG1, NING LI2 AND FENG PENG1
1Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, (Mem. Sun Yat-Sen); Jiangsu province key laboratory for plant Ex-situ conservation, Nanjing, 210014, P.R. China 2Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong SAR, P.R. China
Abstract
This study describes the standardization of an efficient in vitro propagation and hardening procedure for obtaining plantlets from shoot tips of Hemp (Cannabis sativa L.). Hemp seedlings were germinated on half-strength 1/2 MS medium supplemented with 10 g·L-1sucrose, 5.5 g·L-1agar at a pH of 6.8 under light for 16 h per day. MS medium containing 0.2 mg·L-1TDZ, 0.1 mg·L-1NAA supported the maximal auxiliary bud multiplication rate of 3.22 per shoot tip. The proliferated buds were successfully rooted on MS medium supplemented with 0.1 mg·L-1IBA and 0.05 mg·L-1NAA resulting in 85% of the plantlets rooting. The procedure requires a 54 days cycle for the In vitro clonal propagation (14 days for shoot multiplication and 40 days for root induction) which includes 35-42 days for acclimatized plantlet production.

MountZionCollec said:

Thanks Tom, really amazing information thanks for sharing.

You say for storage after propagation but not during, so you take cuttings in flowering for storage this way or did I misunderstand? I've read 45-50 degrees is the temp to shoot for, would you agree? I've read you can root in these conditions, do you root before or just stick the cuttings the medium when placed in Container?

Well my pockets are not fat lol, but enough so after harvest this year that I'm able/willing to invest in my young farm. I misunderstood TC, and appreciate everyone for sharing there information so I didn't make a bad investment!

Click to expand...

You can root in cooler conditions but it will drive your success rate way down.. better to root at usual temps then store in the icebox, yes 45-50f sounds about right.. Even just buried +/- 55f most of the year works really good around my parts.. The goal is to greatly reduce upkeep (growth) as well as all but rid the chance of pathogen. I'd trust this scenario much sooner than i'd trust the current state of this industry - you know how banker's are

Like-ta spend yo money and not give it back.. We are truly sorry mr Holy mt Zion but our policy is we require a one week notice for large withdrawals and we additionally saw fit to place your investment in some various Euro drug futures as well lol...

The real money is opening up call centers in SE Asia to handle the storm of disgruntled investors that is certainly in progress with these types of businesses and going nowhere,,, so solly sir,, daves not here.. no man its me dave kind of thing......

So i hope you are doing all this for the love of the plant, if that is true then i do applaud this,, and yeah,, just bury a container, besides,, how cool is that, right? Right.

TomHill said:

You can root in cooler conditions but it will drive your success rate way down.. better to root at usual temps then store in the icebox, yes 45-50f sounds about right.. Even just buried +/- 55f most of the year works really good around my parts.. The goal is to greatly reduce upkeep (growth) as well as all but rid the chance of pathogen. I'd trust this scenario much sooner than i'd trust the current state of this industry - you know how banker's are

Like-ta spend yo money and not give it back.. We are truly sorry mr Holy mt Zion but our policy is we require a one week notice for large withdrawals and we additionally saw fit to place your investment in some various Euro drug futures as well lol...

The real money is opening up call centers in SE Asia to handle the storm of disgruntled investors that is certainly in progress with these types of businesses and going nowhere,,, so solly sir,, daves not here.. no man its me dave kind of thing......

So i hope you are doing all this for the love of the plant, if that is true then i do applaud this,, and yeah,, just bury a container, besides,, how cool is that, right? Right.

Click to expand...

I grew up amongst corn fields and refineries having never grown a plant.

Because of Marijuana I am free In so many ways, she has connected me to the soil. I love what I'm doing and I am 100% open with anyone on it. I have nothing to hide.

Thank you again my friend, you've probably loved the plant for how long I have lived. Thanks for sharing.