• Title/Summary/Keyword: 육종체계

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Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne) (딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발)

  • Je, Hee-Jeong;Ahn, Jae-Wook;Yoon, Hae-Suk;Kim, Min-Keun;Ryu, Jae-San;Hong, Kwang-Pyo;Lee, Sang-Dae;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.722-729
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    • 2015
  • Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

Introduction of CAX1 into 'Hongro' Apple via Agrobacterium tumefaciens (CAX1 유전자가 도입된 사과 '홍로' 형질전환체)

  • Kim, Jeong-Hee;Shin, Il Sheob;Cho, Kang-Hee;Kim, Se Hee;Kim, Dae-Hyun;Hwang, Jeong Hwan
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.534-539
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    • 2010
  • 'Hongro' is early-mid maturing cultivar with good quality like 'Tsugaru' and it has not preharvest drop. The CAX1 gene was introduced into Korean apple cultivar 'Hongro' by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain transgenic apple with enhanced Ca level. The CAX1 gene playing the role of $H^+/Ca^{2+}$ transporter from Arabidopsis thaliana increases Ca concentration in several plants. Regenerated transgenic lines were confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of CAX1 gene. Southern blot analysis of 'Hongro' transformants showed that two putative transgenic lines were integrated with CAX1 gene in genomic DNA. The CAX1 comparative expression levels of two transgenic lines were higher than that of non-transformant evaluated by comparative quantification analysis using a real-time PCR. These two lines were multiplied in vitro, and micro-grafted on apple rootstocks 'M.9' in the isolated greenhouse. Since two years after micro-grafting, the fruits came into bearing. Compared to Ca level of the non-transgenic 'Hongro', that of the CAX1 transgenic 'Hongro' in the flesh and leaves was higher.

Effective Screening Methods for Lipoxygenase Isozymes in Soybean Seeds (콩 lipoxygenase 효소의 효율적인 검정법)

  • Kim, Young Jin;Park, Tae Il;Cho, Sang Kyun;Oh, Young Jin;Kim, Tae Soo;Kim, Jung Gon
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.26-30
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    • 2008
  • Normal soybean seed contains three lipoxygenase isozymes called L-1, L-2, and L-3, respectively, which are responsible for the generation of undesirable grassy-beany flavors. Simple and effective methods for the detection of lipoxygenase isozymes were developed in soybean seeds. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) has been tried in separating these isozymes. It was done effectively on 7.5% separating gel and 4.5% stacking gel. However, no reliable method has been developed specifically for separating L-3, L-13 and L-23. Visual judging methods were based on the bleaching activities of lipoxygenase in contact with methylene blue and ${\beta}$-carotene. Sodium linoleate bleaching method was adopted to determine L-1 and L-2. Carotene bleaching and spectrophotometric methods were used to determine L-3. These systems were very rapid within one minute, furthermore only required a small piece of cotyledon (below 10 mg) and the other part could be used for generation advance after analysis. It was demonstrated that 200 seed samples could be analyzed per day by one laboratory assistant. The combination of visual judging methods and electrophoresis is suitable for breeding programs. It took 6.5 hours for analysis of 100 seed samples by one person.

Optimization of a protocol for the production of transgenic lily plants via particle bombardment (유전자총 실험조건 최적화를 통한 형질전환 백합 식물체 생산)

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.82-88
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    • 2017
  • Transgenic lily plants have been obtained after particle bombardment, using PDS-1000/He system and scale explants of lilies, followed by PPT (D-L-phosphinothricin) selection. In this study, scales of the lily plants cv. 'red flame' were bombarded with a plasmid containing the bar gene as a selectable marker, and the AtSIZ gene as a gene of interest, showing salt tolerance and drought tolerance respectively, and both being driven by the CaMV 35S promoter. For optimization of a protocol, factors which optimized and showed a high transformation efficiency under following conditions, were considered: a bombardment pressure of 1100 psi, a target distance of 6 cm and $1.0{\mu}m$ of gold particle, and 24-h pre-culture and post-culture on MS medium containing 0.2 M sorbitol and 0.2 M mannitol as osmoticum agents. After bombardment, all the bombarded scales of lily were transferred to MS medium without selective agents, for a week. Subsequently, these bombarded scales were transferred to a selection MS medium containing 10 mg/l PPT, and incubated for a month for further selection, after which they were cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. After transferring into hormone-free MS medium, the PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets. PCR analysis revealed that the surviving putatively transformed plantlets indicated the presence of both the bar gene and the AtSIZ gene. In conclusion, when 100 scales of lily cv. Red flame are bombarded, this study produced approximately 17-18 transgenic plantlets with an optimized bombardment protocol. The protocol described here can contribute to the breeding program of lilies.

Plant Regeneration from Floral Stem Cultures of Nymphoides indica (L.) O. Kuntze. via Somatic Embryogenesis (어리연꽃 (Nymphoides indica (L.) O. Kuntze) 화경 배양으로부터 체세포배발생을 통한 식물체 재생)

  • Oh, Myung-Jin;Min, Sung-Ran;Liu, Jang-Ryol;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.7-10
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    • 2007
  • Plant regeneration system from floral stem of Mymphoides indica via somatic embryogenesis was established. After four weeks of culture onto 1/2MS medium containing 2,4-D, pale-yellow globular structures and calluses were formed on the cut surface of floral stem explants. Upon transfer to 1/2MS basal medium, pale-yellow globular structures were developed into somatic embryos and normal plantlets. These results indicated that pale-yellow globular structures and calluses from floral stem were globular embryos and embryogenic calluses, respectively. The frequency of embryogenic callus formation from floral stem was reached to nearly 100% when floral stem was cultured onto 1/2Ms medium supplemented with low concentration of 2,4-D (0.1 to 0.3 mg/L). However, the higher concentration of 2,4-D resulted in decrease of the frequency of embryogenic callus formation. In this study, low concentration of 2,4-D had a stimulative role in embryogenic callus formation, whereas BA showed inhibitory role in callus formation. In comparison to floral stem, leaf explants showed low frequency of embryogenic callus formation. The highest frequency of embryogenic callus formation from leaf explants was 9.5% when leaf explants were cultured onto 1/2MS medium supplemented with 0.3 mg/L of 2,4-D. The plant regeneration system of Nymphoides indica established in this study, might be applied to mass proliferation, conservation of genetic resources and genetic transformation for molecular breeding.

Meteorological Constraints and Countermeasures in Rice Breeding -Breeding for cold tolerance- (기상재해와 수도육종상의 대책 - 내냉성품종육성방안-)

  • Mun-Hue Heu;Young-Soo Han
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.27 no.4
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    • pp.371-384
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    • 1982
  • Highly cold tolerant varieties are requested not only at high latitute cool area but also tropical high elevated areas, and the required tolerance is different from location to location. IRRI identified 6 different types of cold tolerance required in the world for breeding purpose; a) Hokkaido type, b) Suweon type, c) Taipei 1st season type, d) Taipei 2nd season type, e) Tropical alpine type and, f) Bangladesh type. The cold tolerance requested in Korea is more eargent in Tongil group cultivars and their required tolerance is the one such as the physiological activities at low temperature are as active as in Japonica group cultivars at least during young seedling stage and reproduction stage. With conventional Japonica cultivars, such cold tolerant characters are requested as short growth duration but stable basic vegetative growth, less sensitive to high temperature and less prolonged growth duration at low temperature. The methods screening for cold tolerance were developed rapidly after the Tongil cultivar was reliesed. The facilities of screening for cold tolerance, such as, low temperature incubator, cold water tank, growth cabinet, phytotron, cold water nursery in Chuncheon, breeding nursery located in Jinbu, Unbong and Youngduk, are well established. Foreign facilities such as, cold water tank with the rapid generation advancement facilities, cold nurseries located in Banaue, Kathmandu and Kashimir may be available for the screening of some limitted breeding materials. For the reference, screening methods applied at different growth stages in Japan are introduced. The component characters of cold tolerance are not well identified, but the varietal differences in a) germinability, b) young seedling growth, c) rooting, d) tillering, e) discolation, f) nutrition uptake, g) photosynthesis rate, h) delay in heading, i) pollen sterility, and j) grain fertility at low temperature are reported to be distinguishable. Relationships among those traits are not consistent. Reported studies on the inheritance of cold tolerance are summarized. Four or more genes are controlling low temperature germinability, one or several genes are controlling seedling tolerance, and four or more genes are responsible for the pollen fertility of the rice treated with cold air or grown in the cold water nursery. But most of those data indicate that the results may come out in different way if those were tested at different temperature. Many cold tolerant parents among Japonicas, Indicas and Javanicas were identified as the results of the improvement of cold tolerance screening techniques and IRTP efforts and they are ready to be utilized. Considering a) diversification of germ plasm, b) integration of resistances to diseases and insects, c) identification of adaptability of recommending cultivars and, d) systematic control of recommending cultivars, breeding strategies for short term and long term are suggested. For short term, efforts will be concentrated mainly to the conventional cultivar group. Domestic cultivars will be used as foundation stock and ecologically different foreign introductions such as from Hokkaido, China or from Taiwan, will be used as cross parents for the adjustment of growth durations and synthsize the prototype of tolerances. While at the other side, extreme early waxy Japonicas will be crossed with the Indica parents which are identified for their resistances to the diseases and insects. Through the back corsses to waxy Japonicas, those Indica resistances will be transfered to the Japonicas and these will be utilized to the crosses for the improvement of resistances of prototype. For the long term, efforts will be payed to synthsize all the available tolerances identified any from Japonicas, Indicas and Javanicas to diversify the germ plasm. The tolerant cultivars newly synthsized, should be stable and affected minimum. to the low temperature at all the growing stages. The resistances to the diseases and insects should be integrated also. The rapid generation advancement, pollen culture and international cooperations were emphasized to maximize the breeding efficiency.

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Praziquantel($Distocide^{\circledR}$) in Treatment of Clonorchis sinensis Infection (국산 Praziquantel($디스토시드^{\circledR}$)의 간흡충증에 대한 효과)

  • 서병고;이순형금종일홍성태
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.241-245
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    • 1983
  • PraziquantEI ($Distocide^{\circledR}$), the KcrEan product, was tEstEd for its safety and Efficacy in treatmEnt of Clonorchis sinensiJ infccticn during the period from April to SeptembEr, 1983 in Korea. A total of 55 egg positive cases were selected and treated with the regimen of 25 mg/kg t.i.d. for 1 day (total 75 mg/kg). The follow-up stool examination was done in 47 cases by cellophane thir;k smear and Stoll's egg counting techniques. The 8 uncured cases were treated again with the same regimen. The laboratory tests for blood picture and liver function were done in 27 cases and compared before and after the treatment. The results obtainEd are as follows: 1. After single course treatment, the cure and egg reduction rates were 83.0 and 99.1% respectively. With the second treatment, excellent results of 100% in both rates were obtained. 2. Several kinds of side effects such as dizziness, headache, etc. were complained by 29 cases (61.7 %), however, those were so mild and transient that no special treatment was necessary. 3. No significant change in laboratory findings was recognizable before and after the treatment. From the above results, it is concluded that $Distocide^{\circledR}$ is as effective and safe as $Biltricide^{\circledR}$ and highly recommendable in treatment of C. sinensis infection.

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In Vitro Mitotic Chromosome Doubling by Chemical Treatments in Lilium longiflorum (철포백합의 기내 체세포 염색체 배수화를 위만 화학처리의 효과)

  • Chung, Mi-Young;Chung, Jae-Dong;J. M. VanTuyl;Lim, Ki-Byung
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.73-78
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    • 2004
  • This study was carried out to develop an in vitro chromosome doubling systems for the breeding of tetraploid lily cultivar. Different concentration (ppm) and treatment time (hour) of colchicine, oryzalin and caffeine were compared for the efficiency of bulb regeneration and tetraploidization. The occurrence of tetraploid plants by colchicine was the highest in the concentration of 1,000 ppm for three hours. In oryzalin treatment, the best combination was at 30 ppm for three hours. However, the effects of oryzalin treatments were similar between 10 ppm and 30 ppm concentrations. The survival rate was dramatically reduced in a high concentration of caffeine although some treatments had a higher tetraploid induction. As a consequence, reliable results for the tetraploid production were obtained in the treatments of 1,000 ppm colchicine, 3,000 ppm oryzalin both for three hours, and 9,000 ppm caffeine for nine hours.

Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration of Perennial Ryegrass (Lolium perenne L.) (식물생장조절물질이 페레니얼 라이그래스 (Lolium perenne L.)의 캘러스 유도와 식물체 재분화에 미치는 영향)

  • Lee, Ki-Won;Lee, Dong-Gi;Ahsan, Nagib;Won, Sung-Hye;Lee, Sang-Hoon;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.4
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    • pp.235-240
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    • 2007
  • Optimum tissue culture conditions for an efficient induction of embryogenic callus from mature seeds of perennial ryegrass (Lolium perenne L.) and regeneration of plants from callus tissues were investigated. MS medium containing 3 mg/L 2,4-D and 0.1 mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency (58.3%) was observed when the embryogenic callus tissues were cultured on N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be helpful for molecular breeding of perennial ryegrass through Agrobacterium-mediated genetic transformation.

Molecular cloning, sequences analysis and in vitro expression of the dihydroflavonol 4-reductase gene from Gypsophila paniculata L. (안개초(Gypsophila paniculata L.)로부터 dihydroflavonol 4-reductase 유전자의 분리 및 분석)

  • Min, Byung-Whan;Cheong, Dong-Chun
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.89-95
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    • 2010
  • Dihydroflavonol 4-reductase (DFR) is a key enzyme of the flavonoid biosynthesis pathway which catalyses the NADPH-dependent reduction of 2R,3R-trans-dihydroflavonols to leucoanthocyanidins. In this study we describe cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme DFR in Gypsophila paniculata L. Inspection of the 1279 bp long sequence revealed an open reading frame 1063 bp, including a 36 bp 5' leader region and 181 bp 3' untranslated region. Comparison of the coding region of this DFR cDNA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals an identity higher than 69% at the nucleotide level. The function of this nucleotide sequences was verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants, by in vitro expression yielding and enzymatically active reductase, as indicated by the small leucopelargonidin peak. Genomic southern blot analysis showed the presence of only one gene for DFR in Gypsophila paniculata.