How To Make Stock Solutions For Plant Growth Regulators

Plant growth regulators are an essential component of the culture media because of their role in organ development of the cultured explant. In this video, we are going to explain how to prepare the stock solution of plant growth regulators from their powder forms!
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[@WaltreeNursery]

Thank you for the video. Keep them coming!

[@PlantCellTechnology]

Thanks!

[@MrJvartanian]

Keep up the great videos. They are very helpful.

[@PlantCellTechnology]

Thanks, will do!

[@ceciliavaldovino6110]

Muchas gracias, su video es de gran utilidad.

[@kaminiveer7942]

such a useful video. great job. I would like to suggest moss sterilization.

[@Cheronyx]

Thanks for the video! Can I autoclave TDZ together with medium or not ?

[@laurelnixon183]

thanks for this! not a chem student but I have to do this for my bio experiment. very helpful

[@PlantCellTechnology]

Glad to know that our video was useful to you!

[@RusticRaver]

Hi francisco! quick question; If I wanted to use ethanol as a solvent, how much would you put in a 50ml container? does it evaporate after? thx

[@stevencrowick3803]

i have been a collector and preservationists of strains for 20yrs and have 300 named strains and then the 200 unnamed ones give or take a couple. i am working on RKS from Hermie cheeba world famous IBL heirloom line. One of the most sautés after strain on the market currently, he just releases a few packs after 30yrs of never giving it out to anyone and then died in a motorcycle accident a couple months latter RIP. i want to continue his work of 30yrs. want to start 50k run.

[@rafaelpulido6946]

Thanks for the video! Could you try to propagate begonia from leaf explant?

[@PlantCellTechnology]

Yes, I'll add it to my list!

[@dragonsimon1]

How long can plant growth regulators in powder form last if stored correctly? For example one commonly used 6-Benzylaminopurine (BA), is there a general shelf life? I generally keep the stock solutions for a couple of months before making a new one, but I was wondering what about the powder itself? Appreciate any guidance.

[@PlantCellTechnology]

Powder form hormone can be used until the expiry date of the product (check the bottle to look for the expiry date), provided the opened bottle is stored in a suitable environment.

[@nikkilynick9606]

I would like to see you try the PGR’s with Vanilla orchids. I have watched your starter kit series and videos on how to sterilize in the microwave but still my samples end up contaminated. Not sure what I am doing wrong, I do not have a Laminar Flow Hood so I wear a shower cap, mask and jacket and gloves to try and eliminate particles as well as using 90% alcohol to clean my work area

[@PlantCellTechnology]

tissue culture is a multistep process. contamination can occur because of several reasons, including using unsterilized water, culture vessels, equipment, and tools, improper explant sterilization, stuffed culture room, through the air when the process is not performed under complete sterilized and closed conditions, etc.

[@yungnguyen2311]

Please use on cannabis plants I would love to see what works best. I did not know you had to dissolve it in sodium hydroxide. Thank you I learned something new. Love you videos keep up the great work.

[@PlantCellTechnology]

Thanks for your feedback!!

[@briansmith1544]

You really don’t want to grow cannabis with these hormones unless you really know what you’re doing. They’re great for houseplants, not so much for ingesting…

[@rehab-bi6be]

Great video! I have one question, do you know what solvent is appropriate for meta-topolin (C₁₂H₁₁N₅O) cytokinin?

[@PlantCellTechnology]

Dimethyl Sulfoxide 60 mg/ml

[@rehab-bi6be]

@@PlantCellTechnology Muchas gracias !! 😊

[@joeygiordani8987]

How do you store the powder PGR and liquid PGR? Fridge or room temperature? Thanks!

[@PlantCellTechnology]

Powder can be stored at room temperature and the stock solutions in the fridge.

[@efrenhaw]

I would love to see anthuriums being propagated

[@PlantCellTechnology]

I'll add it to my list!

[@jessesaindon7105]

Thanks for the video! Process question for you. When you use MS Media, do you add the MS vitamins before or after the autoclave/pressure cooker? I've seen it stated that the vitamins should not be autoclaved, but also see that MS Media is often sold with both the basal salts and vitamins combined, which makes me wonder how they are sterilizing everything when it's all all together like that.

[@PlantCellTechnology]

The vitamins included in the traditional MS media can be autoclaved. Some more specialized media are completely made out of scratch, where some of the vitamins are added after autoclave. Some labs also like to add the vitamins, PGRs, and antibiotics after sterilizing the media in the autoclave. Some of those they have to, and the others ones do it because it is their protocol. They use a syringe filter.

[@jessesaindon7105]

@@PlantCellTechnology Thank you!

[@funny11744]

@@PlantCellTechnology What is the size of syringe filter ? 0.25microns ? I understand that Quercetin filtered through 0,45 microns become just half from unfiltered Quercetin. I do not know about other substances.

[@PlantCellTechnology]

@@funny11744 yeah generally 0.22 microns is favored for plant hormones. I didn't get what about quercetin you are asking for. Can you reframe your question?

[@funny11744]

@@PlantCellTechnology I want to mean that after filtration through 0,22 microns It is a big chance that the hormone or any substance to remain lower quantity : the filter itself could retain part of that substance ! I read about Quercetin filtered through 0,45 microns - will remain only about 50% from quercetin .

[@lowatsl6387]

What about macrandra? Which media you could prefer? Could You show sterilization of aquatic mosses?

[@PlantCellTechnology]

1/2 MS media with 1.0 ml/l BA, 0.1 ml/l NAA, 1 ml/l PPM, 25 g of sucrose, and 3 g of gallan gum or 8 g of agar. I'll add aquatic mosses to my list.

[@lowatsl6387]

Mosses are very diffucult. İ Tried 3 4 another ways and no result. Always dead or contamined

[@joeygiordani8987]

Did you sterilize the PGR after you made them? If so how? How long is the PGR solution good for? And how do you store it?

[@PlantCellTechnology]

The PGRs are sterilized with the media in the autoclave/pressure cooker. If they are heat sensitive, with a syringe and a filter after the media has cooled down.

[@imtheheroicdoselive]

Please show how to propagate Homalomena plants.

[@PlantCellTechnology]

Sure. We'll update you if get our hands on Homalomena lants.

[@daniinolte5486]

Good day sir! I have a question: 1. can you sterilize PGR Stock solution in microwave instead of in autoclave? 2. How does syringe sterilization of PGR stock solution work?do you have a video of that? Kind regards

[@theflaca]

Millipore filters that attach to a syringe work fantastic. That's their brand name and they come pre sterilised. just rip open and used in hood.

[@PlantCellTechnology]

Great questions Danii! 1. of course you can sterilize in the microwave. However, you must know the working parameters of your microwave and the knowledge of a suitable temperature and time for sterilization. Also, be on the lookout for how much media you add to each vessel. we have a video on our channel for media sterilization using microwaves. You may find it useful. 2. syringe filter is suitable for plant hormones that heat labile and can not be sterilized. There are easy to use. You have to use a suitable filter with an appropriate filter size for the hormones. directly place them inside the needle, take the hormone and add it to sterilized media. Everything should be performed strictly under sterile conditions, mainly laminar flow hood. Unfortunately currently, we don't have any videos on it. But, we will try to make them in near future soon. Thanks!

[@funny11744]

Regarding 2,4 D =auxin : what is the common concentration in mM or micromoli for every liter of plant media ?

[@PlantCellTechnology]

The concentration depends on what plant you are working on. It's always best to follow a specific protocol for each plant.

[@davidegnesutta3910]

I didn't understand one thing: should we sterilize the pgr stock solution as final step? or only when preparing the media? thank you

[@PlantCellTechnology]

You can sterilize synthetic PGRs with the media. However, filter sterilization is generally required for natural PGRs. A stock solution of hormones is used to directly take the amount of hormone required while preparing tissue culture media. It saves time.

[@farqhankelana220]

how do i use the activated charcoal?

[@PlantCellTechnology]

You weigh the activated charcoal in the required concentration ( gram or milligram per liter, as per your need) and add it to the media. It's similar to the addition of other elements in the media.

[@farqhankelana220]

@@PlantCellTechnology Thank You sir. Do you have any tips on how to sterilize a very small seeds of plant?

[@MrWhiltetail]

You want a challenge? Try Chinese chestnut trees! I would be eternally grateful :-)

[@PlantCellTechnology]

If I can get some seeds.

[@MrWhiltetail]

@@PlantCellTechnology You need seeds to clone a tree? I thought you would need a leaf or stem to clone a tree.

[@MrWhiltetail]

@@PlantCellTechnology Getting seed would not work because then you would be cloning a seedling instead of the mother tree. This would result in loosing the characteristics that I would be trying to replicate. If a seedling was cloned then the protocol may be different for the original mother tree correct?

[@theflaca]

@@MrWhiltetail Try FSAC: fragmented shoot apex culture. I did this in 1989 on grape vines.

[@nabeelz4977]

please propagate Strawberry plants

[@PlantCellTechnology]

Sure! Meanwhile, you can check our blog on the same!

[@carlosdominguez2360]

How do I know if my growth regulators are autoclavable?

[@PlantCellTechnology]

Google search by name. Some companies send documents in which the right way to use the chemical is mentioned.

[@nikkilynick9606]

What can you use if you don’t have any of these plant growth regulators?

[@PlantCellTechnology]

you need PGRs for tissue culture. They are required for plants' growth.

[@nikkilynick9606]

@@PlantCellTechnology where can I buy PGRs?

[@PlantCellTechnology]

@@nikkilynick9606 You can google the websites that sell PGRs. There are many.

[@EdwardGil-gd5pf]

Monstera please, gracias

[@PlantCellTechnology]

Here are some videos that you may find useful: www.youtube.com/@PlantCellTechnology/search?query=monstera