• Title/Summary/Keyword: somatic plant

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Effect of Plant Growth Regulators on Direct Shoots Formation and Somatic Embryogenesis from Leaf Tissue Culture of Muscari armeniacum 'Early Giant' (무스카리(Muscari armeniacum 'Early Giant') 엽절편 조직으로부터 신초형성과 체세포 배발생에 미치는 생장조절물질의 영향)

  • Jeon, Su-Min;Chung, Mi-Young;Lee, Hyang-Bun;Han, Jeung-Sul;Park, Jae Suk;Kim, Chang-Kil;Chung, Jae-Dong
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.4
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    • pp.261-265
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    • 2010
  • This experiment was carried out in order to determine proper plant growth regulators (PGR) and their concentrations for direct shoot induction and somatic embryogenesis from leaf tissue cultures of Muscari armeniacum 'Early Giant'. Direct shoot formation from the leaf explant culture was effective only on a half-strength MS medium containing $0.1mg{\cdot}L^{-1}$ 2,4-D, while embryogenesis was occurred on a half-strength MS medium containing $0.1{\sim}1.0mg{\cdot}L^{-1}$ IPA or without PGR. The regenerated bulblets derived from embryos or shoots were harvested and transplanted into a greenhouse. The sprouting percentage of bulblets obtained from different culture media ranged from 80 to 100% and growth of quality bulblets was enhanced when the bulblets were harvested from the medium containing $0.1mg{\cdot}L^{-1}$ NAA and $1.0{\sim}3.0mg{\cdot}L^{-1}$ IPA.

Development of PCR-based markers for selecting plastid genotypes of Solanum hjertingii (Solanum hjertingii 색소체 유전자형 선발을 위한 PCR 기반 분자마커 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.34-44
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    • 2023
  • The tetraploid Solanum hjertingii, a wild tuber-bearing species from Mexico is a relative of potato, S. tuberosum. The species has been identified as a potential source of resistance to blackening for potato breeding. It does not exhibit enzymatic browning nor blackspot which are physiological disorders. However, due to their sexual incompatibility, somatic hybridization between S. hjertingii and S. tuberosum must be used to introduce various traits from this wild species into potato. After somatic hybridization, molecular markers are essential for selecting fusion products. In this study, the chloroplast genome of S. hjertingii was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop specific markers for S. hjertingii. The chloroplast genome has a total sequence length of 155,545 bp, and its size, gene content, order and orientation are similar to those of the other Solanum species. Phylogenic analysis including 15 other Solanaceae species grouped S. hjertingii with S. demissum, S. hougasii, and S. stoloniferum. After detailed comparisons of the chloroplast genome sequence with eight other Solanum species, we identified one InDel and seven SNPs specific to S. hjertingii. Based on these, five PCR-based markers were developed for discriminating S. hjertingii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary aspects of Solanum species and accelerating breeding using S. hjertingii.

Chloroplast genome sequence and PCR-based markers for S. cardiophyllum (감자 근연야생종 Solanum cardiophyllum의 엽록체 전장유전체 구명 및 이를 이용한 S. cardiophyllum 특이적 분자마커의 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.45-55
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    • 2023
  • The diploid Solanum cardiophyllum, a wild tuberbearing species from Mexico is one of the relatives to potato, S. tuberosum. It has been identified as a source of resistance to crucial pathogens and insects such as Phytophthora infestans, Potato virus Y, Colorado potato beetle, etc. and is widely used for potato breeding. However, the sexual hybridization between S. cardiophyllum and S. tuberosum is limited due to their incompatibility. Therefore, somatic hybridization can introduce beneficial traits from this wild species into the potato. After somatic hybridization, selecting fusion products using molecular markers is essential. In the current study, the chloroplast genome of S. cardiophyllum was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop S. cardiophyllum-specific markers. The total length of the S. cardiophyllum chloroplast genome was 155,570 bp and its size, gene content, order and orientation were similar to those of the other Solanum species. Phylogenic analysis with 32 other Solanaceae species revealed that S. cardiophyllum was expectedly grouped with other Solanum species and most closely located with S. bulbocastanum. Through detailed comparisons of the chloroplast genome sequences of eight Solanum species, we identified 13 SNPs specific to S. cardiophyllum. Further, four SNP-specific PCR markers were developed for discriminating S. cardiophyllum from other Solanum species. The results obtained in this study would help to explore the evolutionary aspects of Solanum species and accelerate breeding using S. cardiophyllum.

Cryopreservation of Embryogenic Tissue and Plant Regeneration in Larix leptolepis (낙엽송 (Larix leptolepis) 배발생조직의 초저온보존 및 식물체 재분화)

  • Kim, Yong-Wook;Kim, Joon-Chul;Youn, Yang;Noh, Eu-Rae;Son, Sung-Ho
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.229-233
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    • 1999
  • The possibility for long-term preservation of Larix leptolepis embryogenic tissue was tested in this study. Higher relative increase of the tissue fresh weight was observed when embryogenic tissue was pretreated for 24 hrs in a medium containing 0.4 M sorbitol or 20% polyethyleneglycol with cooling rate of -0.33$^{\circ}C$/min. The fast cooling rate of -0.5$^{\circ}C$ and -1.$0^{\circ}C$/min appeared to be less effective in regrowth of tissues from cryopreservation. No DNA variants have been observed by PCR analysis among the embryogenic tissues recovered after 1-, 7-, and 28-day-cryopreservation. The post-thaw embryogenic tissue gave rise to mature somatic embryos which developed into plants.

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Assessment of the Glycophorin A Mutant Assay as a Biologic Marker for Low Dose Radiation Exposure (저선량 방사선 노출에 대한 생물학적 지표로서 Glycophorin A 변이발현율 측정의 유용성 평가)

  • Ha, Mi-Na;Yoo, Keun-Young;Ha, Sung-Whan;Kim, Dong-Hyun;Cho, Soo-Hun
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.2
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    • pp.165-173
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    • 2000
  • Objectives : To assess the availability of the glycophorin A (GPA) assay to detect the biological effect of ionizing radiation in workers exposed to low-doses of radiation. Methods : Information on confounding factors, such as age and cigarette smoking was obtained on 144 nuclear power plant workers and 32 hospital workers, by a self-administered questionnaire. Information on physical exposure levels was obtained from the registries of radiation exposure monitoring and control at each facility. The GPA mutant assay was performed using the BR6 method with modification by using a FACScan flow cytometer. Results : As confounders, age and cigarette smoking habits showed increasing trends with GPA variants, but these were of no statistical significance. Hospital workers showed a higher frequency of the GPA variant than nuclear power plant workers in terms of the NO variant. Significant dose-response relationships were obtained from in simple and multiple linear regression models. The slope of the regression equation for nuclear power plant workers was much smaller than that of hospital workers. These findings suggest that there may be apparent dose-rate effects. Conclusion : In population exposed to chronic low-dose radiation, the GPA assay has a potential to be used as an effective biologic marker for assessing the bone marrow cumulative exposure dose.

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Somatic Embryogenesis and Plant Regeneration in Immature Flower Bud Cultures of Carnation (카네이션의 미숙화뢰 배양을 통한 체세포배 발생 및 식물체 재분화)

  • 안병준
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.369-374
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    • 1997
  • Immature flower buds of 'Desio' carnation were cultured on MS agar medium supplemented with 1 ㎎/L 2,L-D. Embryogenic calli were formed from 5-10% of the buds less than 20 ㎜ in length, but only non-embryogenic calli were produced from explants of shoot apex leaf, internode, and flowere buds larger than 20 ㎜. The same method was applied to 16 cultivars of cut Sower carnation and embryogenic calli were obtained in 7 cultivars. Several embryogenic callus lines were selected and maintained through subcultures over 120 weeks without loss of embryogenic competence. The embryogenic cultures were also proliferated rapidly in liquid agitation cultures using MS medium supplemented with 1mg/L 2,4-D. Numerous embryos were formed on the periphery of the cell aggregates upon transfer to auxin-free MS agar medium. Plantlets were transplanted in potting soil and grown to bloom in six months.

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Comparative Cytogenetic Characteristics and Physical Mapping of the 17S and 5S Ribosomal DNAs between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

  • Bang, Kyong-Hwan;Koo, Dal-Hoe;Kim, Hong-Sig;Song, Beom-Heon;Cho, Yong-Gu;Cho, Joon-Hyeong;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.4
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    • pp.311-315
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    • 2003
  • This study was carried out to compare chromosomal characteristics between Atractylodes japonica and A macrocephala. Cytogenetic analysis was conducted based on karyotype analysis and physical mapping using fluorescence in situ hybridization. As a result of karyotype analysis by feulgen staining, somatic chromosome numbers of A. japonica and A. macrocephala were 2n=24. The length. of the mitotic metaphase chromosomes of A. japonica ranged from $0.70\;to\;1.60{\mu}m$ with a total length. of $12.11{\mu}m$ and the homologous chromosome complement comprised six metacentrics, five submetacentrics and one subtelocentrics. On the other hand, the length of the mitotic metaphase chromosomes of A. macrocephala ranged from $0.90\;to\;2.35{\mu}m$ with a total length of $16.58{\mu}m$ and the homologous chromosome complement comprised seven metacentrics and five submetacentrics. The total length of A. japonica chromosomes was shorter than that of A. macrocephala, but A. japonica had one subtelocentrics (chromosomes 4) different from A. macrocepha1a. chromosomes. The F1SH technique using 17S and 5S rDNA was applied to metaphase chromosomes. The signals for 17S rDNA were detected on the telomeric regions of chromosomes 4 and 5 in both A japonica and A. macrocephala. The 5S rDNA signal was found in the short arm of chromosome 1.

Protoplast Culture by Electrofusion of Protoplasts from Solanum sisymbriifolium and Other Solanum Species

  • Kim Hag-Hyun;Jung Hye-Jin;Shin Un-Dong
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.422-426
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    • 2006
  • This research was conducted to get the basic materials necessary to obtain the somatic hybrid plant between Solanum sisymbriifolium and other Solanum species (S. integrifolium and S. toxicarium). Regarding the formation of colony from the protoplast in S. sisymbriifolium, S. integrifolium and the fused protoplast mixture; for the S. sisymbriifolium, a colony was observed in F medium(Kao medium containing $5.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ 2,4-D and $1.0mg{\cdot}L^{-1}$ BA); and for the S. integrifolium, in G medium (a half strength MS medium containing 0.03 M sucrose, 0.4 M mannitol, $1.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ kinetin) respectively. In mixed cultured protoplast after electriofusion treatment, the cell division and colony formation were observed in both media F and G. For the shoot and root formation rate, there was no difference between the parent of each breed and mixed protoplast regardless of the medium. In the fused protoplast mixture of S. sisymbriifolium and S. toxicarium, a colony formation was also observed in both media F and H(a half strength MS medium containing 0.03 M sucrose, 0.4 M mannitol, $1.0mg{\cdot}L^{-1}\;NAA,\;1.0mg{\cdot}L^{-1}$ kinetin); and there was no difference in the shoot and root formation rate between the parent and the mixed protoplast.

Somatic Hybrids by Electro-Protoplast Fusion between N. tabacum and N. glutinosa (담배(N. tabacum)와 N. glutinosa 종간 원형질체 융합식물의 생성)

  • 김준철;최성진
    • KSBB Journal
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    • v.5 no.2
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    • pp.175-182
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    • 1990
  • Protoplasts, isolated from leaf of N. tabacum NR-/SR+ and N. glottnosa were electrofused and divided with a plating efficiency of 30∼35% in AAPI 9M medium. Green callus lines were selected in protoplast-derived colonies on MSNO3 selection medium with 1.2mg/ml streptomycin sulfate on the basis of nitrate reductase proficiency and streptomycin resistance. Four putative hybrid plant lines regenerated from the green callus lines had intermediate morphology between that of parents with respect to floral shape, corolla length and ovate leaf blade. Zymograms of leaf peroxidase and esterase from these putative hybrid plant lines showed isozyme profiles derived from both parents and also, they exhibited additional and lost bands. Cytological analysis of two putative hybrid plant lines gave chromosome counts of 2n=66 in L22 and 2n=54 in L44 which were less than the expected number of N. tabacum(2n=48) and N. glutinosa(2n=24).

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Evaluation of Exogenous Promoters for Use in Brachiaria brizantha Transformation

  • Silveira Erica Duarte;Rodrigues Julio Carlyle Macedo;Cabral Glaucia Barbosa;Leite Juliana de Almeida;Costa Sidnei Souza;Carneiro Vera Tavares de Campos
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.87-93
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    • 2003
  • Brachiaria (Poaceae) is the most important forage genus for cattle production in Brazil. The genetic breeding of this genus is limited by the incompatibility among species, differences in ploidy level and the natural cloning of plants by apomixis (Valle and Miles 1992). However, plant regeneration via tissue culture methods and genetic engineering provide an opportunity to introduce new characteristics in plants of this genus. We have developed methods for the 'genetic modification of Brachiaria brizantha cv. Marandu via biolistic transformation. A higher number of shoots was obtained with 4 mg/L 2.4-diclorophenoxyacetic acid and 0.2 mg/L benzylaminopurine in calli induction medium and 0.1 mg/L naphtaleneacetic acid and 4.0 mg/L kinetin in shoot regeneration medium. A selection curve for mannose was determined to use phospho mannose isomerase (PMI) gene of Escherichia coli as a selection marker. Calli formation was inhibited from 5 g/L mannose, even in the presence of sucrose while calli that were formed in the presence of mannose failed to develop embryos showing that PMI gene can be used for selection of transformants of this grass. Different promoters were tested to evaluate the efficiency based on the detection of the GUS gene expression (Jefferson et al. 1987). The monocot promoters, act1-D and ubi-1, resulted in higher expression levels than dicot promoters, ubi-3 and act-2, or the CaMV35S and CVMV promoters.