Glad to see I'm not the only one using tobacco plants for their landscapingAnd if we make a "bigger" tobacco, then I may need a taller step ladder (with hand rails).
Garden variety allo-tetraploid tobacco (2015).
Bob
Oldfella
Well-Known Member
Why not just lean out of the window? It's safer than the ladder.And if we make a "bigger" tobacco, then I may need a taller step ladder (with hand rails).
Garden variety allo-tetraploid tobacco (2015).
Bob
Oldfella
Tutu
Well-Known Member
It’s been a long time since I started this thread and it kept me busy for a good while. Bob, thanks for opening the thread once again. The idea of making a cross had me very excited for a while but i was too optimistic. At the time i thought that by making a large number of attempts it would sooner or later work out. I guess that i turned from optimist to realist and will now go down a different path.
I have ordered half a laboratory setup. Microscope, still air box, Petri dishes, scalpels, tweezers and a whole bunch of other items.
The plan is as follows. I will make a sexual cross of N. rustica x N. tabacum. Rather than waiting for sheer luck (all previous attempts) i will harvest immature seeds early. I will then grow them in vitro in an MS medium with sugar and agar.
I am sure i will still need to make many attempts before I’ll succeed, but in theory this has a much higher success rate compared to what I’ve been doing before.
After growing various samples in vitro, hopefully i will be able to bring some specimen to adulthood. The next step would be to then get offspring from the plant which will involve some other steps. Earlier in the thread you guys mentioned using cholchicine for doubling chromosomes.
My plan is to get my hands on another herbicide for doing that, Oryzalin.
We’re still far away from those later stages. For now I’m excited for planning the setup of my provisional lab and which crosses i will be attempting. My main goal would be to cross CT Broadleaf as the paternal line and probably go with Aztec, Istela Puebla and Limonka as the maternal lines.
Stay tuned!
I have ordered half a laboratory setup. Microscope, still air box, Petri dishes, scalpels, tweezers and a whole bunch of other items.
The plan is as follows. I will make a sexual cross of N. rustica x N. tabacum. Rather than waiting for sheer luck (all previous attempts) i will harvest immature seeds early. I will then grow them in vitro in an MS medium with sugar and agar.
I am sure i will still need to make many attempts before I’ll succeed, but in theory this has a much higher success rate compared to what I’ve been doing before.
After growing various samples in vitro, hopefully i will be able to bring some specimen to adulthood. The next step would be to then get offspring from the plant which will involve some other steps. Earlier in the thread you guys mentioned using cholchicine for doubling chromosomes.
My plan is to get my hands on another herbicide for doing that, Oryzalin.
We’re still far away from those later stages. For now I’m excited for planning the setup of my provisional lab and which crosses i will be attempting. My main goal would be to cross CT Broadleaf as the paternal line and probably go with Aztec, Istela Puebla and Limonka as the maternal lines.
Stay tuned!
Tutu
Well-Known Member
With some delay on the original schedule, I was finally able to get to it.
Getting the N. rusticas I was growing to flower at the same time as N. tabacum was hard.
Then once they were overlapping, I was away and couldn't get it done.
But I finally had Istela Puebla and Corojo x Sancti Spíritus flowering.
Five days ago after getting back from a trip to Connecticut I managed to open up some young unripe rustica flowers.
I've taken polen from the tabacum and gotten it on there fresh.
This first batch is a bit of a trial. I did cut off all other ripe rustica flowers to avoid cross polination.
With hindsight I should continue to clip flowers maturing days after the fact as well, to avoid chances of cross polination.
So this first batch isn't risk free, but this first batch is mostly set to try out the set-up I have made for early ovule rescuing.
We will get to that in the next post!
* Istela Puebla, it's a stalk grown from a sucker, the original plant I suckered and it had decent size leafs
* Corojo x Sancti Spíritus is also a plant grown from a sucker, the original harvest is still hanging under the stairs and is mostly cured
* Clipped open rustica flowers with stigmas ready to receive some tabacum pollen
* I realize this is tough to make photos off and get into the right perspective
Getting the N. rusticas I was growing to flower at the same time as N. tabacum was hard.
Then once they were overlapping, I was away and couldn't get it done.
But I finally had Istela Puebla and Corojo x Sancti Spíritus flowering.
Five days ago after getting back from a trip to Connecticut I managed to open up some young unripe rustica flowers.
I've taken polen from the tabacum and gotten it on there fresh.
This first batch is a bit of a trial. I did cut off all other ripe rustica flowers to avoid cross polination.
With hindsight I should continue to clip flowers maturing days after the fact as well, to avoid chances of cross polination.
So this first batch isn't risk free, but this first batch is mostly set to try out the set-up I have made for early ovule rescuing.
We will get to that in the next post!
* Istela Puebla, it's a stalk grown from a sucker, the original plant I suckered and it had decent size leafs
* Corojo x Sancti Spíritus is also a plant grown from a sucker, the original harvest is still hanging under the stairs and is mostly cured
* Clipped open rustica flowers with stigmas ready to receive some tabacum pollen
* I realize this is tough to make photos off and get into the right perspective
Tutu
Well-Known Member
So we are five days ahead of time, the ideal day count to rescue unmature ovules from rustica x tabacum cross.
After reading more and more research I have concluded that it is not possible to make this cross naturally.
The cross can be made, and embryos might develop in the ovules, but the plant will abort them.
Therefore, they need to be taken out and grown in a medium outside of the plant.
Most information I've gotten from this paper by N. Shizukuda and T. Nakajima which is called;
Production of lnterspecific Hybrids between Nicotiana rustica L.and N. tabacum L. through Ovule Culture
So over the past weeks I have bought items to get to a somewhat decent setup to take out the ovules and grow it on a medium.
I've used a Murashige & Skoog (MS) Basal Medium called M519, in combination with lab grade agar powder, distilled water and sugar.
* My still air box setup
* After having sterilized all material in a pressure cooker
* First opened the ovaries with a scalpel and then work out the ovules with a pincer
* They weren't hard to find
* It looks a bit slaugheterd but most of the ovules look healthy on all three flowers
* Now the waiting game begins....
After reading more and more research I have concluded that it is not possible to make this cross naturally.
The cross can be made, and embryos might develop in the ovules, but the plant will abort them.
Therefore, they need to be taken out and grown in a medium outside of the plant.
Most information I've gotten from this paper by N. Shizukuda and T. Nakajima which is called;
Production of lnterspecific Hybrids between Nicotiana rustica L.and N. tabacum L. through Ovule Culture
So over the past weeks I have bought items to get to a somewhat decent setup to take out the ovules and grow it on a medium.
I've used a Murashige & Skoog (MS) Basal Medium called M519, in combination with lab grade agar powder, distilled water and sugar.
* My still air box setup
* After having sterilized all material in a pressure cooker
* First opened the ovaries with a scalpel and then work out the ovules with a pincer
* They weren't hard to find
* It looks a bit slaugheterd but most of the ovules look healthy on all three flowers
* Now the waiting game begins....
