Great job sokkos! Your pictures are inspiring me to try tissue culture, but I'll have to do a lot more research before I actually do it. I'd also be interested in some Bletilla striata as well if you have any left.
Got my first Pinguicula to take in tissue culture!
I sterilized leaves from Pinguicula 'Pirouette' and P. gigantea, as well as a single shoot tip from P. 'Pirouette'. All the leaf explants had bacterial contamination (presumably endophytes)after a few days but I managed to get a clean enough explant from the shoot tip. Pinguicula leaves seem to accumulate more endophytes towards the base (pers comm L. Legendre) so I will try to take explants from the tip of the leaf next time instead of using a whole leaf. One week later, once leaves started unfurling from the shoot tip and grew large enough to handle, I separated the leaves and used each intact leaf as an explant for multiplication. I was able to get multiple adventitious buds to form on the leaves by using a protocol from an older paper (Adam et al. (1979) HortScience 14(6):701-702).
I think establishing Pings from explants (and not seed) was probably the largest hurdle for me to overcome; but now that I have clean tissue it looks like smooth sailing from here on. If all goes well, I should have lots of plants to share come fall. Dr. Legendre very graciously sent me a reprint from his article (Legendre (2012) Carniv. Pl. Newslett. 41(1): 16-19) which has an improved medium formulation that I plan to try when I have a bit more time.
One month update: first Pings taken out of tissue culture. Nickel for size reference. The plants in this tray are approximately only a fraction of the total plantlets that regenerated from a single leaf. The speed and volume advantage of tissue culture can really be seen here despite not having optimized the method.
Same tray of plantlets on the left being hardened off enclosed in a plastic baggie. The box on the right shows four clumps of shoots that formed from a single leaf.
Initially, each leaf was covered in adventitious buds three weeks after being put on medium with hormones. To give the buds more space to develop into shoots, each leaf was cut into quarters, hence the four clumps of shoots in the box, and plated back onto the same medium for another three weeks. Once the shoots were big enough to handle, individual shoots were separated and put onto medium without hormones for two weeks before they were big enough to move to soil. All the plantlets had multiple well formed roots 2-3 cm long. In theory it would be possible to produce hundred of plants within a two month time frame.
Very cool thread. It was a pleasure to read it from the beginning.
Are you finished at the UofG? Are you still moving to Manitoba? Is that for work or more education? Will you still be able to play with TC once you move? I hope so, and that you continue to share your accomplishments.
Yes finished at the UofG. I've been in Manitoba for about a year. I moved here for school but definitely intend to move back to Ontario after I'm done. A lot of the plants that I moved over didn't adjust too well and I lost many, but luckily everything in TC survived. Yes I'm still able to play with TC on the side and I've been doing more experimenting (mostly trying out different containers so far, but I'm going to start playing with non-lab grade, supermarket, ingredients to see how cheap I can make TC without compromising efficiency).
dvg: Sarra's safe inside
Mar 31, 2019 20:40:58 GMT -5
bonfield: I've decided to finally reveal my best-kept secret for growing healthy Neps: Just spit on them every few days, the enzymes in saliva help them to better absorb the fertilizer I spray them with!
Apr 1, 2019 14:24:31 GMT -5
dvg: Salivating up your drools?
Apr 1, 2019 16:30:07 GMT -5
dvg: Duped again on April Fool's!
Apr 1, 2019 16:30:45 GMT -5