• Journal of Life Science
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• v.11 no.4
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• pp.340-345
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• 2001

Effects of growth regulators(2,4-D, IAA, NAA, BA and kinetin) and chilling treatment on callus induction. embryogenesis and regeneration through anther cultures of B. falcatum L. were examined. Frequency of callus induction and embryogenesis was effectively increased by the treatment of chilling at 5$^{\circ}C$ for 10 days before anther inoculation. Optimal level of growth regulator for callus induction and embryogenesis from anther was 2,4-D 1.0 mg/L in Murashige and Skoog(MS) basal medium supplemented with 30 g/L sucrose, 8 g/L agar. Frequency of embryogenesis from anther derived callus was increased up to 48% or 45% by addition of IAA 0.1+ kinetin 1.0 mg/L of IAA 0.1+ BA 1.0 mg/L in MS medium, respectively, Optimal medium for obtaining green callus was MS basal supplemented with BA 1.0mg/L. Addition of auxins(IAA or NAA) inhibited the formation of green callus from anther derived callus.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.189-195
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• 1999

This study was carried out to clarify several factors affecting embryogenesis from anther culture of nine cultivars in Brassica oleracea L. var. italica and to investigate the characteristics of plants derived from anther culture. Androgenesis from anther culture was elevated on the B5 medium supplemented with 0.1mg/L NAA, 0.1mg/L 2.4-D and 10% sucrose. Embryo production in liquid medium was five-fold higher than solid medium. High temperature treatment at 35$^{\circ}C$ for one day before transfer to culture room maintained at $25^{\circ}C$ had effective to induce embryogenesis of cultured anthers but extended treatment at 35$^{\circ}C$ decreased significantly the percent of embryogenesis. Frequency of embryogenesis from cultured anthers exhibited significant difference from 2.8% in 'Green Valiant' to 21% in 'Haisi' as affected by genotypes. Percent of spontaneously dihaploid among regenerated plants from anther culture was ranged from 62 to 74% as affected by the genotypes. Characteristic in relation to plant height, number of leaves and branches, and size of head from anther-derived plants showed differential variation in 'Rokguray' and 'Haisi'. Among these charaters obtained from two cultivars, five lines were selected for early maturity, long plant height and large head. Selected lines were used as breeding meterials for F$_1$ hybrid.

• Journal of Plant Biotechnology
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• v.50
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• pp.19-26
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• 2023

Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.5
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• pp.335-340
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• 2002

To evaluate the efficiency of anther floating culture according to the maturing group, the varietal difference and classification of fifty varieties was conducted in N6 liquid medium containing 1mg $l^{-1}$ NAA, 0.25 mg $l^{-1}$ kinetin. The efficiency of callus induction was widely ranged from 0 to 113.4%, but the mean callus induction was not significantly different among maturing groups. The efficiency of anther floating culture showed the highest variation in early-maturing group among three maturing groups. The varieties with the best callus induction were Sambaegbyeo and Jinbuolbyeo, while the recalcitrant variety was Obongbyeo in early-maturing group. The efficiency of plant regeneration showed the highest trends in late-maturing group among three maturing groups. The fifty varieties were classified into three groups (distance=0.78) by cluster analysis based on the callus formation and plant regeneration. Group including only two varieties, Shinunbongbyeo and Sambaegbyeo had the excellent androgenic efficiency, and the medium efficiency of Group was included thirty-six varieties. Whereas twelve varieties, including three Tongil varieties were fell into the bad efficiency of Group. Especially, Tongil varieties containing Japonica rice, Obongbyeo were the recalcitrant genotypes for the anther floating culture.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.33 no.3
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• pp.248-253
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• 1988

The longer pollen stage grew to flowering stage, the higher anther respiratory rate in vivo became. and it was rapidly increased just before flowering. The anther respiratory rate in vitro showed the first and second peak points after 3-7 days and 9-1l days in culture, respectively, and fastest and highest in Daecheongbyeo with high sporophytic potentiality. It was lower in cold-pretreatment than non-treatment at the early days, but higher from 15 days after culture. The frequency of browning anthers was promoted by cold-pretreatment. The respiratory rate was not different between uncolored and browned anthers at 12 days, but it was higher in browned anthers after 24 days in culture.

• Journal of Korean Society of Forest Science
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• v.13 no.1
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• pp.25-39
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• 1971

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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.22 no.3
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• pp.209-212
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• 2002

We obtained regenerated Italian ryegrass (Lolium multiflorum Lam.) plants by anther culture. When Italian ryegrass anther was incubated for 20 days on callus induction medium, MS medium containing 30 g/$\ell$ of sucrose, 2 mg/$\ell$ of NAA and 1 mg/$\ell$ of kinetin, its callus was induced. The ratio of callus induction was 9.2 %, the mean of callus weight was 8.6 mg/callus/anther. When Italian ryegrass callus was incubated for 50 days on plant regeneration medium, MS medium containing 30 g/$\ell$ of sucrose, 1 mg/$\ell$of NAA and 2 mg/$\ell$of kinetin, Italian ryegrass plant was regenerated. The ratio of plant regeneration was 26%.

• Current Research on Agriculture and Life Sciences
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• v.27
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• pp.21-28
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• 2009

The purpose of this study was to improve the culturability of 'IR 36', a indica type rice cultivar using DNA marker associated with the ability of plant regeneration in anther and seed culture. The culturability of 6 rice cultivars and 2 indica/japonica lines ('MGRI 036', 'MGRI 079') were investigated in anther and seed culture. The culturability of 3 japonica rice cultivars were much higher than tongil and indica rice cultivars, and 'MGRI 036' and 'MGRI 079' has high culturability with 20% regenerability, also. 34 $BC_2F_4$ 4 lines were selected by marker screening using RZ400 among 90 $BC_2F_4$ lines derived from a cross between 'MGRI 079' and 'IR 36'. The frequency of callus formation of 10 $BC_2F_4$ lines were higher than 'IR 36' in anther culture among selected 34 $BC_2F_4$ lines. The ability of plant regeneration of 10 lines were higher than 'IR 36' in the seed culture among selected 34 $BC_2F_4$ lines. A promising line, $BC_2F_4$-28, was selected to have better culturability in the anther and seed culture among selected 34 $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4$-28 was similar to 'IR 36'. Using the RZ400 DNA marker associated with the culturability will be useful method for improving of indica rice culticvar's culturability in rice breeding program.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.319-322
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• 1998

Culture conditions for high frequency embryogenesis and plant regeneration in anther cultures of various F$_1$ hybrid and homozygous lines of pepper (Capsicum annuum L.) are described. Anthers pigmented less than halfway from the distal end were dissected from the flower bud in which petals elongated 2 mm higher than the receptacle. They were placed on Dumas medium supplemented with 0.1 mg/L 2,4-D and 0.1 mg/L kinetin. After four weeks of culture, embryos began to appear on anthers. After eight weeks of culture, frequencies of embryo formation reached up to 58.3%. Upon transfer to MS basal medium, greater than 95% of embryos developed into plantlets.

• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.