• Title/Summary/Keyword: Explants

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Plantlet Regeneration Cotyledon and Petiole Cultures of Cyclamen Persicum Mill (시클라멘(Cyclamen Persicum Mill.)의 자엽과 엽병배양에 의한 식물체 재분화)

  • 은종선;고정애;김영선
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.212-216
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    • 1995
  • These experiments were carried out to examine the effect of explant sources and plant growth regulators on callus induction and plantlet differentiation. Cotyledon and petiole explants of Cyclamen persium were cultured on MS medium supplemented with different concentrations of auxins and cytokinins. Cotyledon cultured on medium containing 2, 4-D and kinetin did not form callus or shoots. But when calli induced from petiole explants on medium with 1.0 mg/L 2, 4D and 1.0 mg/L kinetin were subcultured on the same medium the formation of shoots from calli occurred after 150 days of culture. The combination of NAA and BA were more effective than that of 2, 4 D and kinetin in the formation of shoots from calli, cotyledon culture was most effective on medium with 0.2 mg/L NAA and 0.5 mg/L BA. Shoots excised from calli were rooted on medium with 1.0 mg/L NAA. The plantlets were subsequently transplanted to potting soil.

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Expression of Chinese Cabbage Glutathione Reductase Gene in Lettuce (Lactuca sativa L.) (형질전환 상추에서 배추 Glutathione Reductase 유전자의 발현)

  • 정재동;김창길;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.267-271
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    • 1998
  • Cotyledon explants of lettuce were cocultured with Agrobacterium tumefaciens LBA4404::pBKS-GR1 harboring glutathione reductase(GR) gene in MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip for 48 hr. These explants were transferred to MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip, 50 mg/L kanamycin, and 500 mg/L carbenicillin. After 4 weeks of subculture, kanamycin-resistant shoots were obtained on selection medium. Leaves of putative transformants survived on selection medium containing 100 mg/L kanamycin. Incoporation of the GR gene into lettuce was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled GR gene was hybridized to the expected amplified genomic DNA fragment of about 1.8 kb from transgenic lettuce. The presence of mRNA in transgenic lettuce was confirmed by RT-PCR with total RNA of transgenic lettuce. In progeny test of transformants, R$_1$ seeds were resistant to kanamycin (200mg/L) on MS medium.

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Adventitious Root Formation from Cotyledon in Soybean (Glycine max L.) Cultivars (품종별 대두 〔Glycine max L.〕 자엽에서의 부정근 형성)

  • Ha, Keon-Soo;Han, Tae-Jin
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.31-36
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    • 2002
  • The patterns of adventitious root formation from cotyledons for each cultivar of soybeans were compared. The results of adventitious root formation in cultivars are classified as two groups; the first group showed the direct adventitious root formation, and the second group resulted in the callus and adventitious root formation. The cultivars that have much callus formation had less the adventitious root formation. The adventitious root formation in the cotyledonary explants was occured only at the inoculation of adaxial side. When adaxial and abaxial side was inoculated simultaneously, the adventitious roots were formed at the adaxial side. Thus, it suggests that there must be direction to some extent. Starch in the cotyledonary explants were more abundant at the 4 days after induction than at the early stage of the adventitious root formation, but the starch was not observed after 7 days, that the growth stage of adventitious roots.

Distinctive response of maize (Zea mays L.) genotypes in vitro with the acceleration of phytohormones

  • Muppala, Sridevi;Gudlavalleti, Pavan Kumar;Pagidoju, Sreenu;Malireddy, Kodandarami Reddy;Puligandla, Sateesh Kumar;Dasari, Premalatha
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.26-39
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    • 2020
  • In maize, immature embryos (IEs) are highly regenerative explants most suitable for producing high frequencies of plantlet regeneration in vitro. Apart from media, explants, and hormones, genotypic variation also influences in vitro characters to a great extent. In the present study, IEs were used to study the distinctive effect of variation of size/stage and hormones in different genotypes on five in vitro characters viz., frequency of callus induction, growth rate of total callus, frequency of E. callus induction, and volume and number of regenerated plantlets. LS medium with different concentrations of 2,4-D (0.5, 1.5, 2.5, 4.0 and 5.0 mg/L) were used to study the former four in vitro characters, and medium with 6-benzylaminopurine and kinetin (0.5 mg/L, each) was used for plantlet regeneration. IEs of 1.0, 1.5, 2.0, 2.5 and 3.0 mm in size were isolated from four inbred lines viz., NM74C, NM81A, NM5883 and NM5884. Two-way ANOVA revealed that explant size and genotypes, as well as hormonal concentrations showed significant effects on in vitro characters. Two millimeter IEs were found to be suitable for in vitro cultures. LS medium with 1.5 mg/L 2,4-D and LS with BAP and Kn (0.5 mg/L, each) were found to be the best hormonal concentrations for callus induction, maintenance, and regeneration, respectively. Among the four genotypes, NM81A and NM5883 yielded more non-embryogenic and Type I E. calli. In contrast, NM74C and NM5884 yielded more highly regenerative Type II calli. Inbred line NM5884 was found to be the best among these four genotypes.

Multiple shoot induction and plant regeneration from axillary buds of Magnolia 'Vulcan'

  • Kim, Tae-Dong;Kim, Ji-Ah;Lee, Na-Nyum;Choi, Chang-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.40-45
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    • 2020
  • An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia 'Vulcan' was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

Effect of the changes in the Ratio of Dietary Fat to carbohydrate and Energy Restriction on Insulin Sensitivity in Rats (식이중 지방 및 탄수화물의 비율과 열량제한이 흰쥐의 인슐린 민감도에 미치는 영향)

  • 선영실
    • Journal of Nutrition and Health
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    • v.22 no.4
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    • pp.266-274
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    • 1989
  • The current study was undertaken to determine the effects of the ratios of dietary fat to carbohydrate and energy restriction on insulin sensitivity in the growing rats. Male rats weighting 80-90g were fed experimental diets for two weeks. Rats were killed and epiddymal adipose tissue were removed and sliced. Explants of adipose tissues were incubated for 2 hours in KRB(Krebs-Ringer bicarbonate) buffer containing various concentrations of human insulin and [U-14C]glucose. Insulin sensitivity was determined as glucose conversion to total lipids (lipogenesis) during 2 hr incubation. Exp't I : Effects of Ratios of Fat to Carbohydrate on Insulin Sensitivity. Eighteen male rats were fed 3 diets for 2 weeks. Diet 1 was low fat-high carbohydrate (4% soybean oil and 66.5% cornstarch) ; diet 2, medium fat-medium sarbohydrate(12% soybean oil and 58.5% cornstarch) ; diet 3, high fat-low carbohydrate (20% soybean oil and 50.5% cornstarch). Insulin sensitivity was higher in the order of LF-HC, MF-HC and HF-LC diet groups (p<0.05), i.e, lipogenesis was higher at all insuline concentration in the explants from rats fed LF-HC diet. However, thers was no significant difference in body weight gain and epididymal adipose tissue weight among treatments. Exp't II ; Effects of Energy Restriction on Insulin Sensitivity. Twelve rats were grouped into ad libitum feeding and restricted feeding(70% of ad libitum). The experimental diet was medium fat-medium carbohydrat diet as used in the Exp't I. Restricted feeding group tended to show higher insulin sensitivity compared to ad libitum group. However, there was no statistical difference between two groups. As expected, body weight gain and epididymal adipose tissue were higher in the ad libitum group. In summary, the resutls of the current study showed that the epididymal adipose tissue taken from the rats fed low fat-high carbohydrate diet showed higher insulin sensitivity compared to those fed high fat-low carbohydrate, and that resticted feeding tended to elevate insulin sensitivity in these tissues.

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Nano-Encapsulation of Plant Growth-Promoting Rhizobacteria and Their Metabolites Using Alginate-Silica Nanoparticles and Carbon Nanotube Improves UCB1 Pistachio Micropropagation

  • Pour, Mojde Moradi;Saberi-Riseh, Roohallah;Mohammadinejad, Reza;Hosseini, Ahmad
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1096-1103
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    • 2019
  • UCB-1 is the commercial rootstock of pistachio. Reproduction of this rootstock by tissue culture is limited by low levels of proliferation rate. Therefore, any compound that improves the proliferation rate and the quality of the shoots can be used in the process of commercial reproduction of this rootstock. Use of plant growth-promoting bacteria is one of the best ideas. Given the beneficial effects of nanoparticles in enhancement of the growth in plant tissue cultures, the aim of the present study was to investigate the effects of nanoencapsulation of plant growth-promoting rhizobacteria (using silica nanoparticles and carbon nanotubes) and their metabolites in improving UCB1 pistachio micropropagation. The experiment was conducted in a completely randomized design with three replications. Before planting, treatments on the DKW medium were added. The results showed that the use of Pseudomonas fluorescens VUPF5 and Bacillus subtilis VRU1 nanocapsules significantly enhanced the root length and proliferation. The nanoformulation of the VUPF5 metabolite led to the highest root length (6.26 cm) and the largest shoot (3.34 cm). Inoculation of explants with the formulation of the metabolites (both bacterial strains) significantly elevated the average shoot length and the fresh weight of plant compared to the control. The explants were dried completely using both bacterial strains directly and with capsule coating after the three days.

Effect of Medium Composition on in vitro Propagation and Plantlet Regeneration from Nodal Explants of Cassava (Manihot esculenta Crantz)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.16-16
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    • 2019
  • The Cassava (Manihot esculenta Crantz) is a perennial woody shrub cultivated mainly in the tropics for its starchy tuberous roots. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. Thus, a study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, single-node stem segments, approximately 1 cm in length, were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate was 55.6%, the shoot number and its length were 1.0/explant and 2.3 cm in the most favorable 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.

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In vitro micropropagation of two local taro cultivars for large-scale cultivation

  • Alam, Noor Camellia Noor;Kadir, Abdul Muhaimin Abdul
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.124-130
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    • 2022
  • The application of traditional taro propagation methods for large-scale cultivation would be insufficient to meet the high demand for quality planting materials. Therefore, this study aimed to develop an in vitro micro-propagation technique for two local taro cultivars (cv.), Wangi and Putih. Taro cormels were collected from the Malaysian Agricultural Research and Development Institute (MARDI) germplasm (Serdang, Malaysia). Explants were taken from the shoot tip of cormels and initially cultured on Murashige and Skoog (MS) basal media for four weeks. The explants were then transferred to different multiplication media supplemented with different types and concentrations of cytokinins such as 6-benzylaminopurine (BAP ) and Thidiazuron (TDZ). Shoot production was quantified after six weeks of culture. The highest mean number of new shoots was produced by the Wangi cultivar on MS medium supplemented with 2.0 mg/l BAP (2.10 shoots), MS medium supplemented with 0.5 mg/l TDZ (2.18 shoots), and Gamborg B5 medium supplemented with 6.0 mg/l BAP (2.43 shoots). The maximum average number of the Putih cultivar shoots was obtained on MS supplemented with 2.0 mg/l BAP (3.57 shoots). MS basal media was used for root initiation, as it produced an average of 25 roots with an 11-cm length. Various types of substrate mixtures were used during acclimatization. The best acclimatization substrate for the Wangi cultivar was 100% peat soil, whereas the Putih cultivar grew optimally in a combination of peat and perlites at a 1:1 ratio. Taro plantlets require approximately 4 to 6 weeks to acclimatize before they can be transferred to the field.

Cryopreservation of in vitro Grown Shoot Tips of Korean Potato Varieties by Droplet-vitrification

  • Ji-Won Han;Jinjoo Bae;Jae-Young Song;Ho Chul Ko;Sung-Hee Nam;Jung-ro Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.33-33
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    • 2023
  • Potatoes are the world's 4th major food crop after maize, rice, and wheat and also are a staple food for 1.3 billion people. Due to their wide adaptability to various environmental conditions, their yeild capacity, and high commercial value, potatoes have contributed to global food security. Many potato germplasms are commonly preserved as whole plants in fields or in storage to maintain their particular genetic combinations. However, field maintenance is expensive and has the risk of potential losses from diseases, pests, plant ageing and climate change. Over the past four decades, meaningful efforts have been made toward the safe long-term conservation of potatoes through cryopreservation methods such as droplet-vitrification. In this study, we tested 4 Korean potato varieties('Golden Egg', 'Golden Ball', 'Ja-Young' and 'Ha-Ryeong') with the modified potato droplet -vitrification protocol. Potato shoot tips are precultured in a sucrose-enriched medium(0.3 and 0.7M for 7 and 17hrs, respectively) and submitted to a loading step with C4 solution for osmoprotection. The treated explants were dehydrated with Plant Vitrification Solution(PVS)2 which is 80% A3 solution in ice for 30 minutes. Thawing and unloading steps were performed with 0.8M sucrose solution for 30 sec(40℃) followed by 30min(25℃, room temperature). In a potato post-culture medium(MS+0.1 mg·L-1 GA3+0.1 mg·L-1 kinetin), we obtained a survival rates of post-thawed explants ranging 16.1-82.2%. The results suggest that modified and optimized protocols are required dependinig on every cultivar, genetic and ecological types. To achieve higher survival and regeneration rates, each step within the cryoprocedure must be carefully optimized.

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