• Journal of Bio-Environment Control
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• v.8 no.1
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• pp.56-66
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• 1999

Growth of Dendranthema grandiflorum R. ‘Bongwhang’plantlets, as affected by three levels of photosynthetic photon flux (PPF), 70, 150 and 220 $\mu$mol. $m^{-2}$ . $s^{-1}$ , three levels of C $O_{2}$ concentration, 400-500 (ambient), 1000 and 2000 $\mu$mol.mo $l^{-1}$ , and two levels of number of air exchanges per hour (NAEH), 0.1 $h^{-1}$ and 2.8 $h^{-l}$, was studied. Explants were obtained from photomixotrophically-micropropagated plantlets. Four explants were planted in each 3.7$\times$10$^{-4}$ $m^{3}$ polycarbonate box containing MS medium supplemented with 1.25 meq. $L^{-1}$ $H_{2}$P $O_{4}$$^{[-10]}$ and no added sugar. Explants were cultured under cool-white fluorescent lamps (16 h. $d^{-1}$ ), at 25$\pm$1$^{\circ}C$ temperature, and 70-80% relative humidity. In treatments of 2.8 $h^{-1}$ NAEH, a 10 mm round hole made on the vessel cap was sealed with a microporous filter For higher C $O_{2}$ concentrations in the culture room, C $O_{2}$ gas was provided from a tank of liquefied C $O_{2}$. Fresh and dry weights, height, length of the longest roots, number of leaves, and leaf area significantly increased with increasing PPF and especially, with increasing C $O_{2}$ concentration. Growth was enhanced with increased number of air exchanges per hour (2.8 $h^{-1}$ ). Overall, treatment of 220$\mu$mol. $m^{-2}$ . $s^{-1}$ PPF combined with 2000$\mu$mol.mo $l^{-1}$ C $O_{2}$ and 2.8 $h^{-1}$ NAEH gave the most vigorous growth of Dendranthema grandiflorum R. ‘Bongwhang’ plantlets in vitro.o.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.89-95
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• 2000

This study was to establish an effective dilution technique in a vitrification of bovine blastocysts for the field trial. For vitrification, blastocysts were exposed in glycerol (G) and ethylene glycol (EG) mixture in m-DPBS supplemented with 10% FBS. Blastocysts were first exposed to 10% (v/v) G for 5 min, and subsequently were transferred to 10% G plus 20% EG (v/v) for 5 min. Finally, embryos were transferred to 25% G plus 25% EG (v/v) for 30 see and were placed in nitrogen vapor for 3 min, and then were plunged into L$N_2$. At thawing, the straw containing blastocysts was placed in air for 10 sec, and then plunged into a water bath at $25^{\circ}C$ until all ice had disappeared. They were placed in $25^{\circ}C$ and 36$^{\circ}C$ water according to treatment group for different time. Also, in vitro survival was assessed by the re-expansion and hatched rates at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) In the survival rates of vitrified bovine blastocysts according to different dilution time at thawing, the data of 1 min group (86.6, 56.6%) were higher than those of other treatment groups (2 min; 93.5, 35.4%, 2.5 min; 76.9, 30.7%, 3 min; 88.8, 36.1% and 3.5 min; 83.7, 8.1%). 2) When the in vitro survival of vitrified groups according to different developmental stage was examined at 48 h after thawing using 1 min dilution method, the hatching rates of fast developed embryos (expanded blastocyst: 81.3%: early hatching blastocyst: 86.2%) were higher than that of delayed developed one (early blastocyst: 46.6%). 3) In addition, when the in vitro survival of vitrified groups according to different embryo age was compared, the hatched rates at 48 h after thawing of Day 7 (66.6%) and Day 8 embryos (60.0%) were significantly higher than that of Day 9 embryos (22.7%) (P＜0.05). These results demonstrate that vitrified bovine IVM/IVF/IVC blastocysts can be successfully survived in vitro using one-step dilution (1 min) method.