Lilium dauricum is a rare and endangered species belonging to the family Liliaceae. The species contains several bioactive compounds used as functional foods and medicinal agents in Northeast Asia. This study aimed (1) to establish an in vitro bulblet culture using an air-lift bioreactor and callus culture for the conservation of L. dauricum and obtaining its bioactive compounds; (2) investigate the plant phenolic compounds from both cultures system. The highest bulblet production with 12.5-fold increase in growth rate was obtained using MS medium supplemented with 0.5 g L-1 BA and 3% sucrose. Addition of 7% sucrose facilitated bulblet enlargement, with approximate 2.5- and 7-fold increases in diameter and fresh weight, respectively. The highest rate of callus (100%) was obtained using a combination of 1.0 mg L-1 picloram and 0.5 mg L-1 Kinetin. The callus proliferation occurred on MS medium supplemented with 1.0 mg L-1 picloram, 0.25 mg L-1 kinetin, and 0.25 g L-1 casein hydrolysate. There was a significant difference in the total phenolic compound content of callus, which was 1.5-fold higher than that in the bulblets. These findings indicate a suitable system for optimizing both bulblet and callus culture of L. dauricum, therefore, providing useful bio-materials for industrial purposes and contributing to the conservation of this species.
This experiment was to investigate whether the mitochondria function assessment can be used for the prediction of sperm fertility through examining the correlation between mitochondria fluoromicroscopic frequency of frozen-thawed eight Hanwoo bull semen using rhodamine123 (R123) and in vitro embryo development following fertilization. Individual sperm were stained in 5ug/mL R123-added calcium-free Sp-TALP for 30 min at 0 h, 6 h, 12 h and 24 h after thawing and examined their mid-piece under an epifluorescence microscope using 495 nm excitation filter (x1,000). Three replications were taken, and at least 300 sperm per individual were examined. When semen samples were separated into two groups (good and poor) by sperm motility and fluorescent frequencies at just after thawing, average fluorescent frequencies were remarkably reduced as time going (0 h; 53.29~72.94%, 6 h; 21.40~58.90%, 12 h; 8.26~25.93%, 24 h; 1.00~13.78%, irrespective of selected group, and there were no differences at 6 h or 12 h after thawing between selected groups but indicated significant difference at 24 h after thawing (p<0.05). In vitro fertilization rates in good and poor groups ranging 70.8~77.8% and 52.1~84.5%, respectively, were not significantly different. However, in vitro development rates of the same groups ranging 25.7~40.0% and 12.9~1.8%, respectively, were significant different (p<0.05). These results demonstrate that mitochondria fluoromicroscopic assessment of frozen-thawed bovine sperm may be used as a criterion to select more fertile sperm.