General Information
The seed of this species was obtained from plants maintained in my bogs. The original plants were purchased from Shikoku Gardens Nursery in Japan. This plant is found in sphagnum bogs in Japan and Russia. It grows from corms-- no sign of any rhizomes. Roots grow from base of corm. Probably reproduces vegitatively by offset corms. Each stem produces a single leaf and if mature enough a single purple flower. It is easy to cross pollinate and sets seed in a large capsule.
Plating Media: R medium + 2 uM metatoplolin + 4, 1 cm3 cubes Russet potato sliced in quarters + 20 ml/l pineapple 30 mins. 0.525% Chlorox bleaching (no charcoal). Seeds were plated 3/5/2012 on 150 mm petri dishes containing 100 ml of media. Plates were sealed with parafilm and incubated in the dark at room temperature ( approx. 23-26 o C). (Note: The seeds will germinate in the light and produces protocorms with a single shoot).
Number of Days between Plating and Replating: approximately 50 days
Replating Media: R medium + 1, 1 cm3 cube Russet potato sliced in quarters + 20 ml/l pineapple juice (no charcoal) Replating was done in wide mouth pint jars containing 200 ml of media on April 24, 2012. Replates were kept in the dark. Replates were examined on 8/24/2012 after 123 days. The plants consisted of a 1/2 inch stalk (shoot?) terminated in a small bulb like structure at the tip. No signs of true roots were observed.
Number of Days between Replating and Vernalization: 123 days
Number of Days between start and end of Vernalization: ---
Date to Initiate Plating: 10/9
Date to Replate: Nov. 30
Date to start to Vernalize:
Results after Vernalization:
Results After Outplanting in Media:
Notes:
In 2011 success was observed with an alternate protocol which did produce numerous corms.
1. Plated with R medium + 2 uM metatopolin + 4, 1 cm3 cubes Russet potato sliced in quarters, 30 minutes 0.525% chlorox bleaching in darkness.
2. Replating was done after about 26 days on various media. Unfortunately, the media composition was not on the replate jars and the data sheets for the media were missplaced. I tried various concentrations of glucose and NAA and IAA hormones and pineapple juice. In the jars with pineapple juice (low pH) the agar didn't setup and was quite liquid. But there was good shoot and root growth in this media (from memory). So, that is why my current protocol uses pineapple juice.
3. The replates were kept in the dark for 8 months and then kept in the dark in the frig for 3.5 months for vernalization.
4. Then the replates were kept under fluorescent lights at room temperature for 6 months where they produced nice corns, leaves, and long roots.
5. Then the replates were placed back in the frig for further vernalization. Interestingly, the corms at this point did undergo vegetative reproduction to produce more corms. So, I ended up with about 300 small corms. The corms are presently in the frig (Aug. 24, 2012) for further vernalization.