The seed of this species was purchased from Chris Greenwell from Bulgaria. Flower are light purple with some white and don't possess any spots or stripes. The flowers are loosely arranged on the inflorescence not all crunched together. The species is found in wet meadows on alkaline soils.
Plating Media: R medium + 2 uM MetaTopolin + 4, 1 cm3 cubes Russet potatoes sliced in quarters 25 mins. 0.525% Chlorox bleaching (no charcoal). Seeds were plated 11/24/11 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). Plates were examined on Feb. 26, 2012 and germination was poor but adequate enough to produce protocorms for replating. (Note: An alternate media is 1/2X P668 +2 uM MetaTopolin + 4, 1 cm3 potato cubes sliced in quarters 25 mins. 0.525%. chlorox bleaching + activated charcoal. This media was tried and resulted in excellent germination after 30 days but the plate became contaminated and was discarded).
Number of Days between Plating and Replating: approximately 77 days
Replating Media: F522 medium + 1, 1 cm3 cube Russet potato sliced in quarters + 20 ml/l pineapple juice + 0.5g/l charcoal. Replating was done in wide mouth pint jars containing 200 ml of media on March 11, 2012 and replates were kept in the dark. Replates were examined on 8/23/2012 after 165 days. The plants showed good shoot growth with shoots from 1/2 inch to 1 inches with most shoots about 1/2 inches. The protocorms often developed multiple short shoots. The shoots were stout and not spindly. Good root development was also observed but the roots had difficulty penetrating the media. Probably need to reduce the amount of agar for better root penetration. The roots looked healthy and showed no signs of phenolic compounds (this would be hard to see with the presence of charcoal--- but the roots looked excellent). Jars of seedlings were placed in the frig in darkness on Aug. 22, 2012 to start vernalization.
Number of Days between Replating and Vernalization: 171 days
Number of Days between start and end of Vernalization: ---
Date to Initiate Plating: 11/24
Date to Replate: March 11
Date to start to Vernalize: August. 23 (177 days after replating)
Results after Vernalization:
Results After Outplanting in Media: