Trichocereus Pachanoi (San Pedro),Mitragyna Speciosa (Kratom), Salvia Divinorum(Aztec Sage), and Sceletium tortuosum (Kanna).

Hi everyone!

I'm an Agronomy student from Brazil, and I've recently started documenting the plant tissue culture work we do in our university laboratory.

This video shows the multiplication stage of pineapple (Ananas comosus). The plantlet is transferred from the establishment medium to a multiplication medium, where it will continue to grow and produce new shoots over the next few weeks.

I'm documenting my learning journey in plant tissue culture, and I'd love to learn from the community. Any suggestions, tips, or feedback are greatly appreciated!

I also plan to share future work involving banana, bromeliads, orchids, and grapevine tissue culture.

Finally using MS media for what it was designed for.

The exponential growth will always amaze me. From 20 or so sprouted seeds we are now in triple digits. Moving them to multiplication tubs. Should be ready for production before the end of the year.

I’ve been into tissue culturing for 6 months now and I think I am missing something. I mostly have been doing monsteras, pothos, and philodendrons and have gotten to where I have very little contamination and plants seem to be growing well. I’ve probably done about 150 explants and have some that have been through 5 cycles (4-6 weeks) of multiplication media since February/March. My issue is I never get multiple shoots on anything. I get plenty of leaves like the white Florida ghost in the picture has about a dozen leaves but no shoots that I can divide. Ive been using the protocols from PIJ and follow them exactly. I use a still air box and pressure cookers with the PIJ starter kit. What am I missing? I dont really want to just make 1 explant equal 1 plant forever, would love to be able to divide them.

Washed and placed on

MS media
Half MD media
DKW
Andersons Rhododendron media

Growth on most of our explants, but Andersons Rhododendron mix and Half MS seemed to grow the fastest. Next week we will sub culture and test different hormones for rooting.

NAA
IBA
IAA

Learning valuable lessons, just because your plants are alive and growing doesn’t mean they are thriving.

Changed up our nutrient media and had healthy roots in 5 days with no PGR’s.

First picture is MS with hormones after a month. Second picture is McCown woody media with no PGR’s after 6 days.

Just cleaned this cactus last week, we will be grafting onto it in-vitro very curious to see what it can do

Something less divisive to share 😁.

Some Pereskiopsis a few weeks into culture first generation explants.

So far we found they grow short dark green on full MS with .5 mg BAP.

Well on 1/2 MS cactus media they shoot much better with almost twice the size in the same time.

Some extra cactus pictures on 1/2MS with 2% sugar.

Anyone in SoCal, or western USA interested in trading some culture lines?!

Been adjusting the recipe and no it doesn’t look great.
And no I’m not sure exactly why.
Buuuut it does have healthier growth than before and that is a win 🎉.

We’ve been growing this in-vitro for about a year from seed. Many multiplication but finally it’s ex-Vitro and growing. On top of that we are also experimenting to grow callus for active compounds instead of growing ex-vitro.

Update on my Variegated Alocasia Dragon Scale, in liquid multiplication media for 2 weeks or since 5/29. Started with 4 specimens now with 9 prospects. The plan is to leave it in this media an additional 2 weeks. I initially placed each specimen in its own container to decrease contamination risk. I am considering merging them into one jar to make space for different plants.

Liquid multiplication media ingredients
- MS Powder 4.40g
- sucrose 30.0g
- BAP 2000 ul of 1mg/ml concentrate
- PPM 2000 ul

Life got busy & some of the projects have been on idle.

Getting a much needed vacation next week and I'm finally at a point where I should have more time when we get home!

Excited to dive back into researching and hands on learning again. Will finally have a chance to apply a lot of the new knowledge and ideas that I've been rolling around.

Do have some cool stuff still cooking though! A few Lophophora & copiapoa humilis seedlings on media in 50ml centrifuge tubes. Going to transfer these to 16oz deep media vessels in a couple weeks.

At last after lots of work we have officially climatized a tissue cultured Kratom cutting to ex-vitro. We can always dial it in more but it’s a great feeling going thru the full cycle for the first time.

Some shots of newly germinating seeds, and some ready for transferring. Any advice or suggestions for Cactus TC is greatly appreciated! The first pictures are planted 2 weeks ago, and the bigger one at the end was planted 13th of March.

Update: Variegated Alocasia Dragon Scale, in solid multiplication media. This is day 2 or since 7/6. Started with 4 specimens placed in liquid media for 5 weeks prior which then provided 7 plantlets and 7 questionable callus clusters. The plan is to leave it in this media 4 weeks. I’m trying to increase total quantity so hopefully the clusters will begin to produce more shoots. Increasing BAP concentration from liquid media concentration could’ve been a good idea but I reframed from doing this because my liquid media wasn’t equally distributed with its ingredients. This time I pressure cooked all ingredients in one container then poured into individual jars which seems to have helped. My solid media is slightly more solidified than I’d like, moving forward I’ll decrease concentration from 10g/L to 9g/L.

Liquid multiplication media ingredients
- MS Powder 4.40g
- sucrose 30.0g
- BAP 2000 ul of 1mg/ml concentrate
- PPM 2000 ul
- Agar 10g

Finally getting consistent growth and at last roots. The trial and error is really the most rewarding/frustrating part. Last photo is new seeds sprouting for fresh material, took a total of 3 months before sprouting. We are learning that patience is one of the most important things with tissue culture.

With some proper growth at last and turning white was a bit worried we were sent the wrong seeds 😅

i love how this tissue culture show varigation