• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.137-157
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• 1993

This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.215-220
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• 1994

Pollen-derived embryos cultured on the hormone-free medium showed a low germination frequency (12.5%) and poor growth response after germination. The greatest frequency of germination (81.3%) was obtained from the embryos cultured on medium with 0.3mg/L GA$_3$.The greatest frequency of generation (81.3) was obtained from embryos cultured on medium with 0.3mg/L GA$_3$. The embryos precultured for 20 days on medium with 0.3mg/L GA$_3$were transferred to the medium with various combination of hormones such as IAA, kinetin, zeatin, 6-benzylaminopurin (BA) and Gh$_3$. The germination frequency of cotyledonary stage embryos showed above 72% on media with all of the hormonal combinations, but the embryos germinated on medium with 2mg/L BA or 0.1mg/L kinetin and 0.3mg/L GA$_3$ developed more vigorously into plantlets than those of other hormonal combinations. Torpedo-stage embryos cultured on medium with 0.3 mg/L Gh$_3$ were pretreated for 8 weeks at 2-week intervals at 4$^{\circ}C$, The germination frequency of the cold-preheated embryos increased with the increment of pretreatment period from 2 to 8 weeks. The greatest frequency of germination (73.3%) was obtained from the embryos pretreated for 8 weeks at 4$^{\circ}C$. The chromosomes of the root-tip cells of W plane grown for 40 days after germination were observed. Most of the regenerated plants were haploid (55.8%) or diploid (315%), but triploid (1.3%), tetraploid (5.2%), or aneuploid (6.5%) were also detected among them.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.4
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• pp.413-419
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• 1995

This study was conducted to find out the suitable NaCl concentration and temperature for investigating the varietal difference of salinity tolerance in Italian ryegrass. Seeds of 20 cultivars including 9 diploid and 11 tetraploid were exposed to eight levels of NaCl concentration ranging from 0 to 350mM under three temperature levels of 15, 20 and $25^{\circ}C$. The NaCl concentration(C50%) inhibited germination of 50% of the viable seeds for the cultivars at 15$^{\circ}C$ was 354mM, and those at 2$0^{\circ}C$ and $25^{\circ}C$ were 342mM and 325mM, respectively. There were significant correlations among C50% at three levels of temperature, and correlation coefficient for C50% between 2$0^{\circ}C$ and 15$^{\circ}C$ was higher than that between $25^{\circ}C$ and 15$^{\circ}C$ or 2$0^{\circ}C$. C50% was positively correlated only with germination speed at 10 days after sowing when the plants were exposed to 350mM NaCl at 2$0^{\circ}C$. For the germination speed at 10 days after sowing in 350mM NaCl at 2$0^{\circ}C$, the cultivars used could be classified into three groups. Wase Aoba, Bettina, E. K -11, Tetrone, Lira Sand, Tetra Florum and Billiken belonged to the tolerant group, and Tuchi Was, Wase Yutaka, Sakura Wase, Magnolia, Limella and Delita were as the semi-susceptible group, and Atalja, Barmultra, Ajax, Liberta, LM-16, Elving and Wilo were as the susceptible group.

• Journal of Korean Society of Forest Science
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• v.23 no.1
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• pp.9-16
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• 1974

The anthers of late uninucleate microspore or early binucleate microspore stage of Paeonia suffruticosa Andr. (economic tree) were cultured on the modified Murashige and Skoog's medium suppliment with Keinetin, 2,4-D, and NAA for inducing haploid plants. The results are as follows; 1. Callus were induced from both anther locule and anther wall, where that from anther locule was identified as haploid. 2. Among 2,000 anthers cultured, fourteen haploid callus were developed. These haploid callus were clearly identified to be originated from the microspore in anther locule. 3. Diploid callus were induced only 0.5 percent from the callus of endothelium of anther wall, septium of two neighboring anther locule, parenchyma tissues of connectives and or anther locules. 4. In the anther locule of Paeonia suffruticosa Andr. cultured in medium, swollen microspores, polynucleate microspores, multicellurar pollen grains, or callus mass was frequently observed. And the haploids were seemed to be caused by the callus originated from the reduced microspore.

• Journal of Korean Society of Forest Science
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• v.43 no.1
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• pp.6-13
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• 1979

In order to substract the time and cost of propagation for inducing the haploid plants per each species. 500 anthers of late uninucleate microspore on early binucleate microspore stage of Robinia pseudoacacia (Fuel tree) Punius granatum (Ornamental tree). Aleurites fordii (Faty tree) and Styrax japonica (Silvicultural tree) were cultured on the modified Murashige and Skoog's medium supplemented with Kinetin, 2.4-D and NAA as growth regulators. And I observed the samples of cultured anthers under the microscope which were made by Microtoming method and Paraffin method. The results were summarized as follows: 1) Among 500 cultured anthers per each species, anther numbers inducing the diploid callus were as follow: Styrax japonica 20 (4% for the species total); Aleurites fordii 10 (2% for the species, total) and Punica granatum 45 (9% for the species total) were showed. 2) 2n Callus were induced from anther wall. but haploid callus were induced from anther locule. 3) Haploid callus were induced only in 25 anthers (5% for the species total) of Robinia pseudoacacia. 4) These haploid callus were not originated from body cell of anther wall tissue, but from reduced microspores, 5) Since already reported many herbaceous haploid plants were induced from the callus which were originated from reduced microspores, I conclude that the anther of woody plant which induced the haploid callus also will be cultured haploid plant.

• Korean Journal of Plant Resources
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• v.32 no.5
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• pp.509-518
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• 2019

This study was conducted to investigate single crossing condition of M. sacchariflrous and M. sinensis for breeding of M. ${\times}$ giganteus cultivar. Compared with natural day length condition, cultivation in short day length condition shorten days to heading to 18~27 days in both species. Pollen germination ratio of were 75.8% at 6 o'clock in M. sacchariflorus and 51.9% at 7 o'clock in M. sinensis but decreased to below 10% at 8 o'clock in both species. When cut ears immerged in 150 mL of cut-flowers conservation solution and isolated with covering of white non-woven fabric, flowering and pollen dispersal were persisted for 7 days, and the ratio of pollen germination were above 40% for 4 days. The ratio of self-fertilization of both species were below 2.5%, but open pollenation ratio were above 50%. We obtained 437 seeds with experimental single cross of 14 combinations between tetraploid M. sacchariflorus and diploid M. siensis by application of developed single crossing methods. In the single cross, numbers of seed set were different by mother plants. Thus, the newly investigated single crossing condition will be used to breed M. ${\times}$ giganteous cultivar which is sterile and has superior characteristics of biomass yield.