FlytrapCare Carnivorous Plant Forums

Doomsday wrote:I'm just using 1/3MS, and testing out different hormones and combinations of them

As I understand it, hormones shouldn't be used on seedlings.

Matt wrote:

Doomsday wrote:I am growing some dionaea seeds on callus media, and instead of growing like yours, they are turning in to little blobs:

I've seen a few people do this over the years and in all cases I can remember, the seedlings never start growing correctly after germinating on hormones.

Matt wrote:

Doomsday wrote:I'm just using 1/3MS, and testing out different hormones and combinations of them

As I understand it, hormones shouldn't be used on seedlings.

That's interesting. I figured if I could get them to callus straight away. it would be easier than growing them out for a bit, and then taking explants from them, and inducing callus on those. If that can't be done, that sucks, but I don't see why it couldn't... A callus should be the same whether it's made from seedling cells or mature plant cells, no? Either way, the callus cells should contain all of the plant's dna, and be "controllable" via hormone application.

I was planning on letting these get to green pea/kidney bean size, and then cutting them in half, and putting half back on callus media, and half on hormone-free media, to try to get them to grow into bunches of plants via embryogenesis. I got the idea from seeing a few people's seeds turn straight into callus after germination. I'm not sure if those calli were hormone induced, or just on plain media, though.

I did start my Drosophyllum lusitanicum seeds and my D. nitidula gemmae on hormone-free media, to see how it is multiplying them after letting them put on some size.

Doomsday wrote:That's interesting. I figured if I could get them to callus straight away. it would be easier than growing them out for a bit, and then taking explants from them, and inducing callus on those.

I think you're misunderstanding how propagation in tissue culture works. Flytraps propagate just fine from seeds without any hormones. Just sow them on hormone free media, let them grow and replate as they grow. In a year or so you will have hundreds of them.

Doomsday wrote:A callus should be the same whether it's made from seedling cells or mature plant cells, no? Either way, the callus cells should contain all of the plant's dna, and be "controllable" via hormone application.

I don't know the details, but when a plant is emerging from a seed, it seems that giving it hormones at that point disrupts its growth. From what I've gathered in reading books and online, it's fairly common knowledge in the TC world to not use hormones on seedlings. And all my personal experiments and others I've seen have always resulted in plants that don't grow well. One person replated them for several months, perhaps a year or so, after starting Dionaea seeds on hormones and the plants never showed normal growth so he just threw them out.

It looks like you have a great germination rate, and barely any contamination.
How do you treat the seeds against contamination before placing them in the media?

Matt wrote:

Doomsday wrote:That's interesting. I figured if I could get them to callus straight away. it would be easier than growing them out for a bit, and then taking explants from them, and inducing callus on those.

I think you're misunderstanding how propagation in tissue culture works. Flytraps propagate just fine from seeds without any hormones. Just sow them on hormone free media, let them grow and replate as they grow. In a year or so you will have hundreds of them.

Doomsday wrote:A callus should be the same whether it's made from seedling cells or mature plant cells, no? Either way, the callus cells should contain all of the plant's dna, and be "controllable" via hormone application.

I don't know the details, but when a plant is emerging from a seed, it seems that giving it hormones at that point disrupts its growth. From what I've gathered in reading books and online, it's fairly common knowledge in the TC world to not use hormones on seedlings. And all my personal experiments and others I've seen have always resulted in plants that don't grow well. One person replated them for several months, perhaps a year or so, after starting Dionaea seeds on hormones and the plants never showed normal growth so he just threw them out.

I'm aware that certain species like Dionaea divide pretty rapidly in-vitro, but I want to get good at callus culture as well, so I can use calli for both long-term storage, and propagation of plants that don't divide as well by themselves. I also want to get good at callus induction so I can induce them on explants instead of only starting from seed.

I'm gonna try replating some of these on hormone-free media soon, to see what happens. It would be pretty annoying for them not to eventually grow normally, but at the very least, I learned that my media recipe for callus induction works pretty well. I'll be sowing some of my next batch of dionaea seeds on hormone-free media, to compare.
Thanks for your input, @Matt. You've got me searching for an explanation for such phenomenon, now.

YaronR wrote:It looks like you have a great germination rate, and barely any contamination.
How do you treat the seeds against contamination before placing them in the media?

Thanks! I've been experimenting with NaDCC, instead of using the traditional bleach treatment. I've had some contamination, likely bacterial, but the solution I've been using is much gentler on seeds/tissue than regular bleach treatments, and doesn't require any rinsing, either. I've also been experimenting with adding in a little Polysorbate20 (tween20) as a surfactant.

Drosophyllum #1 Update:


Drosophyllum #2 Update:

Doomsday wrote:Thanks for your input, @Matt. You've got me searching for an explanation for such phenomenon, now.

If you find some explanation, pleasepost here so Iknow too!

Matt wrote:

Doomsday wrote:Thanks for your input, @Matt. You've got me searching for an explanation for such phenomenon, now.

If you find some explanation, pleasepost here so Iknow too!

Will do! These plants never cease to amaze me. There's always something new to be learned about them. It still blows me away how such a ridiculously small dose of hormones can cause such a drastic change in how they grow.

Droso seedling doing really well after being given proper lighting:

Here are some pieces of D. regia callus I cut in half and then replated.... I couldn't tell how soft they were, until they were all plated up already. That's what I saw that they had broken apart, somewhat. I moved them from callus inducing media to hormone-free media. I wonder if anything will come of them. Hopefully they aren't contaminated, so I can find out.

D. sp. "Floating"

Before:


After:

Dude. What the heck is that turning into? I swear I see a paw growing now.