• Title/Summary/Keyword: in vitro rooting

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Effect of growth regulators on In Vitro mass propagation of Haworthia maughanii (생장조절제가 하월시아 만상(Haworthia maughanii)의 기내 대량증식에 미치는 영향)

  • Kim, Youn Hee;Kim, Hye Hyeong;Lee, Gee Young;Lee, Jae Hong;Jung, Jae Hong;Delgado-Sanchez, Pablo;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.369-374
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    • 2018
  • The purpose of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shoot and rooting for the in vitro mass production of Haworthia maughanii. To determine suitable parts of the plant for callus induction, the leaves, flower bloom and flower stalks were cultured in MS medium at different concentrations of $0{\sim}2mgL^{-1}$ NAA and $0{\sim}2mgL^{-1}$ TDZ, respectively. All of the parts showed 100% callus formation rate at $NAA\;1mgL^{-1}$ and $TDZ\;1mgL^{-1}$ treatment, $NAA\;2mgL^{-1}$ and $TDZ\;2mgL^{-1}$ treatment and NAA 1 to $2mgL^{-1}$, respectively. While the rate of callus formation was high in all parts, the leaves were the most efficient to obtain most culture parts. $NAA\;0.1mg\;L^{-1}$ and $BA\;0.1mg\;L^{-1}$ treatments were the most effective in shoot formation with 22.0 shoots. In addition, multiple shoot propagation showed 16.3 shoots, the highest, with $NAA\;0.1mg\;L^{-1}$ and $BA\;0.1mg\;L^{-1}$ treatments. These results led us to speculate that the optimization of culture conditions was responsible for the mass propagation for in vitro cultures of Haworthia maughanii.

Effect of kinds of medium and plant growth regulators for plantlets regeneration by bud culture in Disporum sessile (윤판나물(Disporum sessile) 아배양에 의한 식물체 재분화에 영향하는 배지 종류 및 생장조절물질 효과)

  • Lee, Na-Nyum;Kim, Ji-Ah;Kim, Tae Dong;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.42-48
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    • 2017
  • This study was conducted to establish the optimal condition for in vitro propagation of Disporum sessile. In the experiments with different kinds of media (MS, B5 and WPM) and explant types for shoot induction and elongation, the highest number of shoot inductions (2.5/explant) was shown when the axillary bud explants were cultured on MS medium without plant growth regulators (PGRs). The best shoot elongation (7.2 cm) was also obtained when the apical buds were cultured on MS medium. The effect of BA pulse treatments with in vitro shoots was also examined. The highest in shoot induction (2.29/explant) and elongation (7.28 cm) was observed when the shoots from axillary buds were cultured on the media without PGRs. However, it decreased with increasing duration of BA pulse-treatment. The highest rooting rate (100%) and number of root inductions (21.3/explant) were achieved with 1.0 mg/L indole-3-butyric acid (IBA) treatment, whereas no differences were observed by different shoot types. The regenerated in vitro plantlets were hardened and successfully established in soil.

Micropropagation of Cucurbita foetidissima and Asclepias syriaca through Shoot Tip Culture of Seeding (Cucurbita foetidissima 와 Asclepias syriaca의 정단배양을 통한 기내 대량증식)

  • 염미란;이선교;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.63-69
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    • 2000
  • In order to establish a micropropagation system for buffalo gourd (Cucurbita foetidissima ) and common milkweed (Asclepias syriaca), the effects of several plant growth regulators and culture temperature on shoot multiplication and rooting were investigated. In buffalo gourd, the greatest number of shoot from shoot tip culture and well growth of formed shoot were obtained on the MIS medium supplemented with 1.0 mg/L BA and 0.3 or 0.6 mg/L IAA. Whereas kinetin and 2iP were not effective for shoot multiplication in vitro. It was found that 22$^{\circ}C$ and $25^{\circ}C$ were suitable for shoot multiplication. Roots were easily formed by the addition of auxins, especially 1.0 or 2.0 mg/L IBA and 2.0 mg/L IAA. Over 90% of plants survived successfully after being transferred into the field. In common milkweed, BA was more effective than kinetin or 2iP for its micropropagation in vitro. The increased shoot weight and number of nodes per shoot were obtained on the medium containing 3.0 mg/L BA and 0.3 or 0.6 mg/L IAA. But 2iP promoted the shoot elongation. In addition. common milkweed was sensitive to culture temperature in vitro. Temperature around 22$^{\circ}C$ was favorable for shoot multiplication and growth, whereas temperature higher than $25^{\circ}C$ usually reduced the rate of shoot survival rate.

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In Vitro Micropropagation of Polygonatum odoratum cv. Gungangbeaksea (둥굴레 '건강백세'의 기내 대량번식)

  • Kim, Man Bae;Park, Chun Geun;Yi, Sookyi;Han, Bong Hee
    • Korean Journal of Plant Resources
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    • v.27 no.5
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    • pp.540-545
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    • 2014
  • The Polygonatum odoratum cv. Gungangbeaksea, bred in Gyeongsangnam-Do Agricutural Research & Extension Service, was cultured in vitro for micropropagate rapidly through the culture of rhizome explants ($5{\times}5mm$). The $7{\times}7mm$ explants of adventitious multi-bud clusters (AMC), obtained through the culture of rhizome explants (MS + 3.0 mg/L BA) were cultured on MS media with BA and TDZ. The shoot multiplication was favorable on the MS medium containing 3.0 mg/L TDZ with 2.8 in shoot number. But the formation of AMC was low in all media tested. The explants of AMC were cultured on MS media containing 1.0~5.0 mg/L TDZ and NAA to multiplicate AMC more. The formation of AMC was a little more stimulated on combined MS media of TDZ and NAA, than that with TDZ alone. The multiplication of shoots and AMC was favorable on MS media with 3.0 mg/L TDZ and 5.0 mg/L NAA, and 5.0 mg/L TDZ and 3.0 mg/L NAA. As the concentration of MS salts increased, the formation of AMC was decreased. But the formation of AMC was more stimulated, as the concentration of sucrose increased to 7%. Therefore, the multiplication of shoots and AMC was suitable on media containing 3.0~5.0 mg/L TDZ and NAA, and 7% of sucrose. The explants of AMC were rooted on media with 3.0 mg/L IBA, or 2.0 mg/L NAA with more than 80% in rooting ratio. The plantlets were treated at $5^{\circ}C$ for 8 weeks, and cultured ex vitro for 8 weeks. The survival ratio of plantlets were 100% in vermiculite, and the mixed soil with perlite 1 volumn and vermiculite 1 volumn.

In vitro Regeneration and Genetic Stability Analysis of the Regenerated Green Plants in Japanese Blood Grass (Imperata cylindrica 'Rubra') (홍띠 기내 재생과 재생 녹색식물체의 유전적 안정성)

  • Kang, In-Jin;Lee, Ye-Jin;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.34 no.2
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    • pp.156-165
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    • 2021
  • The in vitro regeneration was established, and the genetic stability among the mother plants (control) and the micropropagated green plants was evaluated using ISSR markers in Imperata cylindrica 'Rubra', Poaceae which containing important bioenergy plants. Green shoots were multiply induced from growing point culture via callus on MS medium supplemented with 0.01 mg/L NAA and 2 mg/L BA, and the shoots were proliferated on the MS medium with rooting. Rooted plantlets were transplanted to the pot with 100% survival rate. Using ISSR markers, somaclonal variation was analyzed in eight mother plants (control), ten green-regenerant cultivated at culture room (ReR) and ten green-regenerant cultivated at field condition (ReF). All ISSRs produced a total of 97 bands, and the scorable bands varied from one to seven with an average of 4.4 bands per primer. The polymorphism rate of ReRs and ReFs was 4.1% and 3.1% respectively, showing higher rate than that of control (0%). The genetic similarity matrix (GSM) among all accessions ranged from 0.919 to 1.0 with a mean of 0.972. According to the clustering analysis, ReFs and mother plants were divided into two independent groups. The results indicate that no clear genetic diversity was detected among regenerated plants, and ISSR markers were useful tool for identification of somaclonal variation of regenerants.

High-frequency regeneration by stem disc culture in selected clones of Populus euramericana

  • Cui, Hae-Yeon;Lee, Hyo-Shin;Oh, Chang-Young;Han, Shim-Hee;Lee, Kyung-Ju;Lee, Hyun-Jeong;Kang, Kyu-Seok;Park, So-Young
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.236-241
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    • 2014
  • An efficient regeneration protocol for stem disc culture of Populus euramericana, which is important species for bioenergy resource in agroforestry, was established. The number of explants that were obtained and the number of explants that regenerated varied with the genotypes. However, in all the genotypes, stem disc culture produced more regenerated shoots than did in axillary bud culture. A comparison of the effects of cytokinin type and concentration on shoot regeneration in different explants (i.e., petiole, leaf, and root segments of P. euramericana) revealed that a concentration of $0.002mg\;l^{-1}$ thidiazuron (TDZ) used on petiole segments resulted in the greatest shoot regeneration (95.83%). The hormonal requirements for the greatest shoot regeneration in the three explant types varied. Different concentrations of $AgNO_3$ and $CoCl_2$ were added separately to the medium to stop the yellowing and subsequent necrosis of the regenerated shoots. Lower concentrations (3 and $5mg\;l^{-1}$) of these compounds improved shoot regeneration and elongation, compared with the control. The in vitro-regenerated shoots were transferred to rooting medium and subsequently acclimatized. The highly efficient regeneration system of P. euramericana reported here can be used for mass propagation of this recalcitrant for regeneration, economically important tree species.

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.

Micropropagation of the hybrids of Actinidia deliciosa$\times$A. arguta by tissue culture (참다래$\times$다래 교잡종의 액아배양 및 캘러스 배양에 의한 기내번식)

  • 문흥규;권영진;이병실
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.227-230
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    • 2001
  • Kiwi (Actinidia deliciosa) is exotic plant and thus susceptible to cold climate in the middle part of Korean peninsular. Several hybrids have recently been developed to enhance cold tolerance by crossing them with domestic species (A. arguta), We have developed an efficient micropropagation technique for the hybrids using both axillary bud and callus culture systems. Shoot proliferation from axillary buds was possible on St medium supplemented with 0.2 mg/L Bh and 3.0 mg/L GA$_3$. In vivo cuttings of the proliferated shoots were more effective for root induction and subsequent survival than in vitro rooting. More than 95% of the plantlets were successfully transferred to field. Effective callus induction was achieved on MS or B$_{5}$ medium with 2,4-D or NAA. Although callus induction could be made from any combinations of media and auxins, shoot regeneration was observed only from the callus induced on medium containing NAA.A.

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The change of somatic cell embryogenesis in Kalanchoe pinnata because of agar concentration in stimulating root stress (뿌리 스트레스를 유발하는 agar농도에 따른 Kalanchoe pinnata의 체세포 배 형성 변화)

  • Park, Jongbum;Kim, Jin-Seok;Kim, Donggiun
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.320-324
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    • 2017
  • Development of modern agricultural machinery and accompanying agricultural development cause soil compaction and reduce growth by stressing roots. Kalanchoe pinnata was used to investigate the impact of stress on rooting and changes in plant growth and reproduction. K. pinnata forms somatic embryos capable of asexual reproduction at the edge of leaves. Impact of root pressurization of K. pinnata on somatic embryogenesis and organ differentiation according to external stress factors was investigated by using a high concentration of agar and this phenomenon was studied histologically. Agar concentration in culture media ranged from 0.5%-1.5% to induce a compression effect on roots. The stem and leaf of K. pinnata were subjected to a microtechnique process to study changes in tissue. In vivo, K. pinnata produced 2nd and 3rd plantlets at edges of leaves from lack of water and excessive lighting conditions. In in vitro culture studies, the lower the concentration of agar, the higher the population and the higher the biomass, but plantlet did not occur in leaf bends. Conversely, as concentration of agar increased, increase in the number of individuals was low. Plantlet development occurred only in agar 1.5% medium. The difference in agar concentration was a stressor in the root of K. pinnata, and thus the pattern of asexual reproduction changed from the division method in root to a plantlet generation in leaf. This suggests root pressurization may act as stress and change in the plant reproduction pattern.

The Protein and Isozyme Patterns During in vitro Plant Regeneration of Yooja (Citrus junos Sieb.) and Trifoliate Orange (Poncirus trifoliata Rafin.)

  • Park, Min-Hee;Jang, Hyun-Kyu;Cha, Young-Ju;Kim, Ho-Bun;Lee, Sook-Young
    • Plant Resources
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    • v.5 no.1
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    • pp.29-44
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    • 2002
  • In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

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