• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.23-23
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• 2021

Apple (Malus×domestica Borkh.; Rosaceae) is an important fruit crop grown mainly in temperate regions of the world. Tissue culture in vitro is a biotechnological technique that has been used to genetically improve cultivars (scions) and rootstocks. This could be important in the production of genetically uniform scions and rootstocks for commercial apple production. In nurseries, apple plants are produced by grafting scions onto rootstocks. The Cornell-Geneva (Geneva® series) breeding program has bred several dwarf rootstocks that are resistant to diseases and pests and are also cold hardy. This study was conducted to determine the optimal medium strength to improve sprouting shoot rate of apical meristem of the apple rootstocks of fire blight resistance. The apple rootstocks apical meristem at size (0.2 mm to 0.3 mm) with axillary buds were cultured on the MS(Murashige & Skoog) medium supplemented with plant growth regulators. The sprouting ratio and growth characteristics was evaluated after eight weeks in vitro culture. The highest rate of bud differentiation and shoot formation were 23.8% and 55.6%, respectively. After 6 weeks, shoots were regenerated from apical meristem, and their growth characteristics was significantly varied on the respective basal medium with different plant growth regulators. Our studies showed that the apple rootstocks the apple rootstocks of fire blight resistance plantlets could be successfully produced from apical meristem differentiated out of young twigs via organogenic regeneration.

• Korean Journal of Plant Resources
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• v.28 no.6
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• pp.690-696
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• 2015

We micropropagated pear (Pyrus species) using shoot tips and nodal explants from three pear genotypes. The ability to establish shoot tip cultures, proliferate shoots, induce rooting, and acclimatize the resulting plantlets are all elements of in vitro micropropagation. Shoots were induced from shoot tips on Murashige and Skoog medium (MS) with five different plant growth regulator combinations. The highest shoot formation rates were achieved for the three genotypes using MS supplemented with 1.0 mg/L N⁶-benzyladenine (BA) and 0.1 mg/L gibberellic acid (GA₃). The maximum shoot number and shoot length for the three cultivars were recorded with 2.0 mg/L BA and 0.2 mg/L indole-3-butyric acid (IBA) in multiplication medium using nodal explants produced from microshoots. Nodal explants with one or two axillary buds cultured for three weeks initiated roots on medium supplemented with various concentrations of 1-naphthaleneacetic acid (NAA) or/and IBA in half-strength MS medium for adventitious rooting. The highest rooting response was with the combination of 0.2 mg/L NAA and 0.2 mg/L IBA. A combination of NAA and IBA resulted in a significant increase in the rooting ratio over NAA or IBA alone. In this medium, the root formation rate according to ranged from 68.9% for the BaeYun No. 3 genotype to 51.8% for the Hwanggeum genotype. We also investigated the influence of the concentration the polyamine phloroglucinol in rooting medium. For all three genotypes, the highest rooting ratio, longest root length, and greatest root number were observed in the treatments with 75-150 mg/L phloroglucinol. Most rooted plants were acclimatized successfully.

• Journal of Plant Biotechnology
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• v.2 no.1
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• pp.21-24
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• 2000

An efficient method of shoot regeneration of peanut is described. In vitro shoot organogenesis from the callus of cotyledon explants of Arachis hypogaea L. was stimulated by addition of Adenine sulphate (Ads) along with 6 - benzylaminopurine (BAP) and - napthalene acetic acid (NAA). Ads (13 ${\mu}{\textrm}{m}$) had a stimulatory effect on shoot bud differentiation when combined with BAP (13 ${\mu}{\textrm}{m}$) and NAA (2 ${\mu}{\textrm}{m}$). Shoot organogenesis was markedly higher (92%) from callus induced on Ads, BAP and NAA combined media than from those formed by the individual supplementation of Ads or BAP with NAA. The shoots elongated on the media with GA$_3$ (1 ${\mu}{\textrm}{m}$). Elongated plantlets rooted with MS media containing IBA (9 ${\mu}{\textrm}{m}$).

• Korean Journal of Plant Resources
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• v.11 no.1
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• pp.1-8
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• 1998

This study was conducted to establish mass propagation system from the axillary bud culture of chrysanthemum zawadskii H. which was used as material of medicinal plants. Shoot egeneration was better on MS medium with NAA and BA. The optimum concentraions of growth regulator for shoot regeneration differed depending on accessionsof C. Zawadskii. Shoot regeneration in Keungucheolcho was better on MS Medium with NAA 0.01mg/1 and BA 0.1mg/1 while Hyangrobonggucheocho was better with NAA 0.1mg/1and BA 0.3mg/1. Addition of NAA into medium was effective for induction of root from shoots regenerated. Shoot multiplcation was more effective when 10mg/1 spermine was added into medium than when other polyamines were treated ino medium . Randomly and specifically amplified polymorphic DAC banding patterns based on polymerase chain reaction (PCR) analysis were used to assess the genetic variation of plants regenerated from in vitro culture.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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• pp.50-50
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• 2020

본 연구는 천궁에 감염된 오이모자이크바이러스(CMV)와 천궁엽맥황화바이러스(CnVYV)의 제거를 위해 항바이러스제인 리바비린 처리 효과를 조사하기 위해 수행되었다. 천궁 기내식물체의 생장점을 절취 후 항바이러스제 농도를 달리한 배지에서 생육을 유도한 후 RT-PCR 방법을 통해 바이러스 검출율을 조사하였다. 리바비린을 40 mg l^-1 농도로 처리하였을 때 바이러스 검출 종 수는 1.1종으로 감소하였다. 또한 동일조건에서 CMV와 CnVYV-2의 검출율은 무처리구 대비 각각 1/5과 1/2 수준으로 감소하였다. 하지만 리바비린 처리에 따른 CnVYV-1의 검출율은 감소하지 않았다. 리바바린 처리 농도에 따른 생존율은 처리구별로 유의적인 차이가 나타나지 않았으나, 40 mg l^-1 리바비린 처리구에서 신초 발생 수와 신초 길이는 무처리구 대비 절반 수준으로 감소하였다. 리바비린 처리를 통해 CMV와 CnVYV-2가 미검출된 23개체를 선발하였으며, 추가적인 바이러스 검정 단계를 거쳐 무병묘 생산에 활용될 것이다.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.145-150
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• 1994

This study was conducted to determine the effects of 3 cytokinins (BA,2iP and kinetin) and their concentrations (0, 0.1, 0.5, 1.0, and 2.0 mg/L) on multiple shoot production of Citrus spp. 'Sambokam' and 'Byungkyool' by nodal culture. Nodal explants were obtained from in vitro germinated seedlings of both cultivars. 'Sambokam' produced more multiple shoots than did 'Byungkyool' by nodal culture. Among the 3 cytokinins tested in this study BA supplemented in semi-solid MS basal medium was the most effective stimulator for multiple shoot production, and an optimal concentration was determined to be 1.0 mg/L. Shoot elongation and root formation were inhibited by increasing cytokinin concentration, regardless of cytokinin types. BA at 1.0 mg/L produced the most multiple shoots and the highest number of leaves in 'Sambokam', whereas any cytokinin and concentration studied in this experiment did not affect any scored variables such as shoot and leaf numbers, etc. in 'Byungkyool'.

• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.145-149
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• 2004

We investigated the anatomical aspects of vitrification in peace poplar. Comparisons were made with regard to characteristics occurring between hyperhydric and normal shoots in shoot proliferation cultures on MS medium containing 0.2 mg/L BA. Compared with normal plants, hyperhydric plants had thick, curled, and dark green leaves. Hyperhydric stems were thicker and shorter than those of normal stems. When examined under the microscopes, the mesophyll palisade cells of hyperhydric leaves were vacuolated, whereas those of normal leaves contained normal and enriched vacuole with cytoplasm. Generally, the hyperhydric leaves showed poorly developed palisade parenchyma, and revealed irregular and bigger sized intercellular structures in both palisade and spongy parenchyma as well as epidermis cells compare to those of normal leaves. In addition, the hyperhydric leaves had lower stomatal density and bigger sized cell. Vascular tissues of hyperhydic stems were less differentiated because of poorly lignified xylem tissue. The greatly expanded cortical cells and pith appeared to be the main cause of thick stems as compared with normal stems.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.131-135
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• 1994

This experiment was designed to improve the in vitro production system of apple rootstock M.26 as being influenced by the growth regulators, TDZ, BA, IAA, IBA, zeatin, and GA$_3$. Different levels of potassium humate (KH), known as cytokinin and auxin-like substance, were also supplemented to the MS basal medium along with IBA 0.6 mg/L to find out it effect on root formation in apple rootstock M.26. ID initiate and establish the in vitro multiplication of shoots byway of meristem culture, MS medium added with zeatin 1.0 mg/L was found to be the most suitable, showing the 100% of survival rate of shoot tips. A combination of thidiazuron (TDZ) 0.2 mg/L and NAA 0.5 mg/L promoted the shoot proliferation when shoot tips were used as explants. MS basal medium plus IBA 0.6 mg/L was very effective for root induction, but an addition of potassium humate (250 mg/L) to the medium containing IBA 0.6 mg/L stimulated the induction and proliferation of the rook by far the better.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.24-24
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• 2021

The Cassava (Manihot esculenta Crantz) is one of the major food crops in the tropical or subtropical regions. Recently, clean planting materials of improved cassava cultivars are in high demand. Problems in the propagation of cassava are virus vulnerable and low rates of seed germination. Thus, the study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. So we tried to optimize protocols for mass production from axillary buds of Cassava. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with axillary buds, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, axillary buds approximately 1 cm in length were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate showed 55.6%. The shoot number and its length was 1.0/explant and 2.3 cm in the most favorable medium composition. The auxin β-indolebutyric acid(IBA) 0~2.0 mg/L was proved to be effective on root development. Plantlets with fibrous roots easily generated tuberous roots in vitro. The tuberous roots were induced only when both kinetin and IBA were used in combination. after 8 weeks, the root generation rate showed 100%. The root number and its length was 17.2/explant and 2.2 cm in the most promising medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

• Plant Biotechnology Reports
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• v.5 no.2
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• pp.147-156
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• 2011

Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.