Thuja koraiensis Nak. is a short and creeping evergreen shrub which reaches about 3 m in height and only occurs in the northeast China and in high mountains over the Korea. It's designated as a rare and endangered tree species in Korea and DD (data deficient) in Red List Category & Criteria of IUCN. This study was carried out to develop the propagation technique by cutting for conservation of genetic resources of T. koraiensis. The rooting responses of branch cuttings, obtained from hard (May) and semi-hard wood shots (August) to three plant growth regulators (PGRs), namely, IAA, IBA, and NAA applied at various concentrations (0, 100, 500, 1000, 2000, and 3000 mg/l) were examined in sand and mixed soil media. Percentage of rooting showed significant difference between cutting time, among kinds and among concentration of PGRs. The optimum cutting time was April to May in hardwood cutting. The application of IAA 1000 mg/l and NAA 500mg/l were effective in callus formation and rooting of cutting. Relatively, rooting of cutting of the control taken in May was above 93%.
Germination percentage of Korean native lily seeds was high at $20-25^{\circ}C$. It was almost 100% in L. cernuum, L. callosum, L. amabile, and L. concolor, 88.0% in L. lancifolium, and 73.0% in L. maximowitzii, respectively. Meanwhile, it was low rate of 34.0%-54.0% in L. distichum, L. hansonii, and L. tsingtauense. Germination was mostly delayed of $15^{\circ}C$ and days to germination were more shortened in species with higher germination percentage. Even though the effect of daylength was not considerable in germination rate, it was promoted in L. maximowitzii but it was delyed in L. hansonii under long day. The effect of soaking in hot PGRs solution in L. callosum, L. cernuum, L. amabile, L. lancifolium, and L. concolor did not show any difference in comparison with non-treatment. However, it was improved by BA in L. maximowitzii. Longer period of cold wet storage resulted in improved germination percentage in L. maximowitzii and L. lancifolium, while it affected decreased percentage in L. distichum and L. hansonii. Days to germination were shortened by longer period of cold wet storage regardless of species. Germination percentage in dry storage was higher under cold temperature than room temperature and under desiccator storage than outside desiccator, it was highest under desiccator storage at $4^{\circ}C$. It was drastically reduced by the non-use desiccator storage at room temperature L. concolor, however it was improved only by the use of desiccator L. maximowitzii for a long time.
This experiment was conducted to evaluate the effect of plant growth regulators, dichlorprop and MCPB on the reduced effect of fruit drop and fruit quality before and after storage in apples. Dichlorprop was tested with dilution of 1000 at 30, 40, 50 days before harvesting, and MCPB with dilution of 4000 at 15, 25, 35 days before harvesting. The results are summarized as follows : Percentage of fruit drop was appeared to the notable reduction as compared with the untreated control when regulators was applied with dilution of 1000 at 30 days before harvesting by dichlorprop and with dilution of 4000 at 35 days before harvesting by MCPB. Degree of fruit colour showed to the remarkable promotion at all the treatment of 30, 40, 50 days before harvesting by dichlorprop as compared with the untreated control. Sugar contents in flesh was increased a little at the treatment of 30 days before harvesting by dichlorprop, but acid contents in flesh was reduced at all the treatment of 30, 40, 50 days before harvesting by dichloroprop and at 15, 25, 35 days before harvesting by MCPB. Passed firmness of fruit after storage was maintained at the treatment with dilution of 4000 at 35 days before harvesting. Therefore, it was repressed a softening of fruit, but by dichlorprop treatment at 30, 40, 50 days before harvesting, fruit firmmess was appeared to reduce according to the passage of storage period. Amount of ethylene evolution after storage was showed to reduce at all the treatment by early treated time of dichoroprop and MCPB, but carbon dioxide increased at treatment conditions such as the front. Accordingly, these relationship showed to be contrary each other.
The objective of this study was carried out to investigate the proper plant growth regulator for increasing the number of flower, fruit set, and to enlarge the size of the berries in Ardisia pusilla. Flower bud formation was used rooted cutting, and fruit set, enlargement, and coloration of fruit were used with two years-old. $GA_3$ concentrations were treated with 0, 100, 200, or $400mg{\cdot}L^{-1}$. Flower bud formation was effective in $400mg{\cdot}L^{-1}$$GA_3$ and it was 1.8 times greater than control. Plant growth regulators were applied by foliar spray at full bloom stage to increase the fruit set. As a result, $GA_3$ was the most effective for increasing fruit set. Also, auxins of 4-CPA (Tomatotone, Donbu hitech Co., Korea) and dichloprop triethanol amine (Antifall, Bayer Crop Science Co., Ltd., Korea) were effective. When $GA_3$ concentrations of 0.5 and $1.0mg{\cdot}L^{-1}$ were used, fruit set (%) reached to 70% and 77%, respectively. Effectiveness of $GA_3$ was 1.8 times greater than control. Also, auxins, dichloprop triethanol amine increased to about 7-12% during fruit setting, but cytokinin and anti-gibberellin were ineffective. To investigate the fruit enlargement and coloration, $GA_3$ was treated with 0.3, 0.6, and $1.2mg{\cdot}L^{-1}$. Fruit enlargement was achieved to about 15% by $GA_3$$0.6mg{\cdot}L^{-1}$ when $GA_3$ was treated 3 times at the interval of 1 month per treatment when fruit size was about 2-3mm (after full-blooming two months). But anthocyanin contents for coloration of fruit skin were not significant according to $GA_3$ concentration. The results showed that $GA_3$ enhanced bud formation, fruit set and enlargement of fruit size in Ardisia pusilla.
Nerine from south africa and its sparkling flower shape make us estimate it as a hopeful kind of cut follow. There was a few studies on Nerine in korea. We started this study to set bulb propagation methods. The propagation by tissue culture was changeable according to the growth regulators The best growth regulator combination which makes a lot of Bulblet was NAA $0{\sim}0.5$ + BA $0.5{\sim}2.0mg\;{\cdot}\;L^{-1}$ in Nerine bowdenii ‘Favourite’ and Nerine sarniens ‘Red’ respectively. The adjust culture media source for tissue culture were glucose 9% as a carbon source and ($NH_4+NO_3$) 40mM as a nitrogen source. When glucose was used as a carbon source, Bulblet were harvested a little bit low then sucrose but comparative emergence rate was so high that it is good for carbon source in nerine tissue culture. When we consist culture media as MS+BA $1.0mg\;{\cdot}\;L^{-1}$+sucose 7% + ($NH_4+NO_3$) 40mM, the produced Bulblet were reached up to 1.7 each per bulb and emergence rate was up to 100% irrespective of acclimatization period. The suitable culture explant for nerine tissue culture was scale. When scale was cultured with MS+BA $1.0mg\;{\cdot}\;L^{-1}$+sucose 7%, its propagation efficiency was 54 times greater than using growing point. A proper culture part of the scaly leaf was middle part (8 scaly leaf from outer 8th scaly leaf) when middle part was cultured the number of Bulblet were up to 1.8 each per explant.
Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.
Recently successful induction of haploid plant by means of anther culture method has become a big topic among geneticists and plant breeders. The haploid plant can be used as a precious material for such basic researches as mutation or genetics. Once the haploid is obtained, production of homozygous plant is not a difficult problem. The method of producing homozygous plant can, also, be applied to the practical breeding works. When applied to the hybridization of self-fertilizing breeding period would be greatly shortened and in cross-fertilizing vegetables production of uniform hybrid seed would be very easily obtained. Last few years many scientists attempted anther cultures using various plant species, but it was successful only in several species. Unlike the other tissue cultures which use somatic organs or tissues as explants, anther culture seems to be very difficult because the plants or calli have to be induced from the haploid microspores or pollen grains. In the present experiment anther culture of fruit trees and ornamental shrubs of four genera and seven species was attemped. Anthers of Various stages ranging from tetrad and late microspore were cultured on the modified Murashige and Skoog's medium supplemented with various concentrations of auxins and kinetin as growth regulators. Handling of materials, sterilization, and other operations of culture were done by routine methods. The results were summarized as follows: 1. Calli were induced in the anthers of Forsythia Koreana Nak., Rhododendron mucronuratum Turcz., R. yedoense Max. var. Poukhanense Nak., and Prunus armeniaca L. var. ansu Max. No signs of callus were observed in Prunus persica Sieb. et Zucc. var. vurgaris Max., Pyrus ussuriensis var. macrostipes (Nak.), and Prunus salcina Lindley. 2. Calli were easily formed in any of the media with differing concentrations of auxins and kinetin. 3. In F. Koreana calli developed from anther surface and connective. Callus emerging out of anther locule was not observed. 4. Somatic calli arose from filament, connective, and inside of anther wall in R. mucronulatum. Many of the microspores accumulated starch grains. 5. The anther lobes located opposite the filament of R. yedoense turned easily to calli. This phenomenon was not observed in R. mucronulatum. Microspore embedded for a period in the medium became starch pollen. No callus was observed arising from microspore. 6. In P. armeniaca calli were not induced from somatic anther tissues. Instead, callus emerged out of anther locule rupturing the anther slit. Starch was not formed in the microspore. 7. In P. persica, Pyrus ussuriensis, and P. salcina, calli were not observed in the anthers examined more than 60 days after culture. Microspores of these species, however, were free of starch grains even after long period of subculture. 8. It was learned that somatic calli of the species examined arose usually from endothelium of anther wall, septum of two neighboring anther locules, parenchyma tissues of connectives, or anther lobes. 9. In the anther locule of P. armeniaca cultured long in medium, swollen microspores, polynucleate microspores, multicellular pollen grains, or callus mass were frequently observed, this indicating that the callus of this species was microspore-origin. 10. It was clarified that in P. armeniaca production of haploid plant by anther culture might be possible.
Seo, Mi-Suk;Sohn, Seong-Han;Park, Beom-Seok;Ko, Ho-Cheol;Jin, Mina
Journal of Plant Biotechnology
/
v.41
no.3
/
pp.116-122
/
2014
Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.
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.
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.
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