• Title/Summary/Keyword: in vitro cultured petiole

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High Frequency Plant Regeneration from Leaf Explant Cultures of Domestic Cultivated Strawberry (Fragaria x ananassa Duch) (국내 딸기 재배품종의 잎절편 배양으로부터 고빈도 식물체 재생)

  • Cho Mi-Ae;Choi Kyu-Myeong;Ko Suck-Min;Min Sung-Ran;Chung Hwa-Ji;Liu Jang-Ryol;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.175-179
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    • 2005
  • To develop a high efficiency plant regeneration system from in vitro cultures of strawberry, cv. Yeobong, petiole and leaf explants were cultured on MS basal medium containing a combination of 0.5 mg/L IBA and 3.2 mg/L kinetin or zeatin or benzyl amino purine (BAP) for 6 weeks, and leaf explants with dark pretreatment for a week ($T_1$), 2 weeks ($T_2$), and 4 weeks ($T_3$) were cultured on medium supplemented with 0.5 mg/L IBA and 3.2 mg/L zeatin under 16 hr photoperiod for 6 weeks. Shoot organogenesis was observed from the greenish calli containing minimal anthocyanin formed at proximal cutting edges of the leaf explant (57%) when cultured adaxial side on the medium, whereas was directly formed from a cutting edges of petiole explant (6.3%). Frequency of callus formation and shoot organogenesis at large size of leaf explant ($1.0{\sim}1.5\;cm^2$) was higher than small size ($0.5{\sim}1.0\;cm^2$), and dark pretreatment significantly improved the frequency of leaf explants that produced calli and shoots. The maximum frequency (87%) for shoot organogenesis was obtained from the leaf explants that transferred to a 16 hr photoperiod condition after the initial 4 weeks dark period. The improved frequency (87%) in comparision with control without dark pretreatment (27%). When the shoots were transferred to 1/2 MS basal medium, formed roots with 20 d of culture. The rooted plants were subsequently transferred to the pots and to the field.

High Frequency Plant Regeneration from Leaf, Petiole and Internode Explants of Codonopsis lanceolata Benth.

  • Ghimire, Bimal Kumar;Shin, Chul-Min;Li, Cheng Hao;Kim, Na-Young;Chung, III-Min;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.73-81
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    • 2007
  • An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 $mg/{\ell}$ 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 $mg/{\ell}$ NAA and 1 $mg/{\ell}$ BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

Influence of plant growth regulators on adventitious shoot formation of Glehnia littoralis Fr. Schmid (갯방풍의 기내부정아 형성에 미치는 식물생정조절물질의 영향)

  • Choo, Byung-Kil;Ji, Yun-Ui;Moon, Byeong-Cheol;Choi, Go-Ya;Lee, Hye-Won;Lee, A-Yeong;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.113-116
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    • 2008
  • Petiole explant of Glehnia littoralis Fr. Schmidt was in vitro cultured MS plant medium(DUCHEFA co.) supplemented with various plant growth regulators and examined to find out their optimum combination and concentration for plantlet regeneration. We investigated optimal condition for efficient plant regeneration through adventitious shoot formation on MS plant medium with various kinds of plant growth regulators. Embryogenic calli and adventitious shoot formation were greatly influenced by plant growth regulators. Embryogenic calli induction showed a good response on MS medium supplemented with NAA and BA than others. Especially, combination of 1.0 mg/L NAA and 0.5 mg/L BA on MS medium led to the greatest frequency in adventitious shoot. The results suggest that plant regulator selection be important factor to achieve an efficient regeneration.

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Callus induction and high-efficiency plant regeneration via somatic embryogenesis in Papaver nudicaule L., an ornamental medicinal plant

  • Yang, Jing Li;Zhao, Bo;Seong, Eun-Soo;Kim, Myong-Jo;Kang, Won-Hee;Kim, Na-Young;Yu, Chang-Yeon;Li, Cheng Hao
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.261-267
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    • 2010
  • We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

Studies on the Mass Propagation of Pinellia ternata (Thunb.) Breit in Vitro (조직배양에 의한 반하〔Pinellia ternata(Thunb.) Briet〕의 대량번식에 관한 연구)

  • 최정식;나의식
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.31 no.1
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    • pp.30-42
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    • 1986
  • In order to find out the best media, explants and environmental conditions for induction of calluses and organogeneses of Pinellia ternata (Thunb.) Breit in vitro, various parts of adult have been cultured on Murashige & Skoog's medium containing various levels of 2,4-dichlorophenoxy acetic acid(2,4-D) and kinetin. The results obtained were as follows: Calluses were induced from the surface of apical meristem and leaf tissue. Formation and growth of calluses in petiole ex plants were best on the MS medium complemented with 2,4-D 2.0 mg/l and kinetin 0.2mg/l. But callus formation in stem ex plants of the nearest tuber was not induced at all kinds of media. Plantlets occured at all treatment except absence of growth regulator. Their numbers, size, leaf and fresh weight were promoted by 2,4-D 2.0mg/l and kinetin 0.2mg/l. Root growth was increased on the medium containing higher 2,4-D concentrations. Size and fresh weight of callus were increased at 25$^{\circ}C$ compared with 10, 20 and 30$^{\circ}C$, respectively. Optimal pH value was at 6.0 for growth of callus. Morphological aberrations were observed in plantlets, especially in regenerated leaves. The separation of the broad leaved plantlets and albino were observed in some cultures. Growth of plantlets after transplantation was best in pots with the sterilized vermiculte. But abnormal variants withered up.

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Callus and Micro-Crown Bud Formation in Vitro from Leaf Explant of Yacon (Polymnia sonchifolia Poeppig & Endlicher) (야콘 (Polymnia sonchifolia Poeppig & Endlicher) 잎의 절편체로부터 캘러스 및 기내 소관아 형성)

  • 두홍수;권태호;박철형;류점호
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.101-107
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    • 2000
  • The explants of yacon (Polymnia sonchifolia Poeppig & Endlicher) were cultured to invest th8e dedifferentiation condition, and formative callus from leaf was cultured to find the regeneration and micro-crown bud formation. Basal MS medium was more effective to form callus than 1/2 MS and B$_{5}$ medium. Calli formations from leaf, petiole and lateral bud were more effective on MS medium supplemented with 1.0, 2.0 mg/L 2,4-D and 0.2, 0.4 mg/L kinetin or BA than 1.0, 2.0 mg/L NAA and 0.2, 0.4 mg/L kinetin or BA. Formative callus from leaf was proliferated about 70% on medium supplemented with 1.0 mg/L BA. When callus was proliferated, 63% regeneration rate was shown on medium supplemented with 1.0, 2.0 mg/L BA in case of subculture for 3~4 months but was not shown on medium supplemented with 1.0, 2.0 mg/L kinetin. Micro-crown bud formed as addition of BA at 3~4 months after callus culture and then was obtained many at 5~6 months, it was most formed about 82% on medium supplemented with 5 mg/L BA. Rate of micro-crown bud formation was increased as more over 5 mg/L BA concentration, when this time, however, shoot had thick leaves and short internodes, and then withered before long, Micro-crown bud was formed about 88.0% on medium supplemented with 5% sucrose, that was more increased 28% than with 3% sucrose. The buds of crown bud between harvested in field and formed in vitro were difference only in size, but both were similar in shape according to histological view.

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Effects of Several Culture Conditions on in vivo Growth and Development in Gerbera hybrida (기내 배양환경이 거베라 유묘의 기외이식 후 생육에 미치는 영향)

  • 이현숙;임기병;정재동;김창길
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.91-95
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    • 2001
  • Propagules grown at different in vitro culture conditions such as heterotrophic, mixotrophic and photoautotrophic conditions were investigated for growth, total photosynthesis ratio and flowering. Survival rate of propagules after transplanting was higher in photoautotrophic propagules than in the heterotrophic and mixotrophic ones. Total photosynthesis was higher plantlets growth in photoautotrophic (154 mg$CO_2$.mgDW$^{-1}$ h$^{-2}$ ) those grown than in mixotrphpic (148 mg$CO_2$.mgDW$^{-1}$ h$^{-2}$ ) and heterotrophic (102 mg$CO_2$.mgDW$^{-1}$ h$^{-2}$ ) 30 days after transplanting into fields. Day to flowering of the plant cultured in photoautotrophic condition was shortened by 7~10 days than those of heterotrophic and mixotrophic ones. Length of the petiole, number of leaves, leaf area and chlorophyll content were also increased.

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Establishment of high frequency plant regeneration system from leaf explants of Pinellia koreana via bulblets formation

  • Oh, Myung-Jin;Park, Jong-Mi;Lee, Bu-Youn;Choi, Pil-Son;Tae, Kyoung-Hwan;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.193-196
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    • 2009
  • Pinellia koreana K-H Tae & J-H Kim is a recently discovered Korea endemic medicinal plant species whose natural habitat is rapidly destroyed by industrial development. Described in this paper are culture conditions for high frequency plant regeneration via bulblet formation from leaf explant cultures of P. koreana. Leaf explants formed white nodular structures and off-white calluses at a frequency of 91.2% when cultured on MS medium supplemented with 2 mg/L BA and 0.5 mg/L NAA. However, the frequency of white nodular structures and off-white calluses formation was slightly decreased with an increasing concentration of NAA up to 4 mg/L, where the frequency reached 31.7%. Most petiole explants did not form white nodular structures and off-white calluses except the combination treatment of 2 mg/L BA and 2 mg/L NAA. Upon transfer onto MS basal medium, over 90% of nodular structures gave rise to numerous bulblets and developed into plantlets. Plantlets regenerated from bulblets were transplanted to potting soil and grown to maturity at a survival rate of over 95% in a growth chamber. Therefore, the in vitro plant regeneration system of P. koreana obtained in this study will be useful for mass propagation and long-term preservation of genetic resources of P. koreana.

Characterization of In Vitro Totipotency by Armoracia rusticana (서양고추냉이의 기내 전형성능에 관한 특성)

  • BAE, Chang-Hyu;MIN, Kyung-Soo;AHN, Chang-Soon;LIM, Yong Pyo;KAMEYA, Tosiaki;Lee, Hyo-Yeon
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.119-124
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    • 1997
  • Plantlets were regenerated from various explants (shoot tip, leaf blade, petiole and root segments) via organogenesis and/or somatic embryogenesis from Armoracia rusticana(Lam) Gaerth., Mey, et Scherb.. Shoot regeneration rate from callus was highest on the MS mediums supplemented with 0.5 ㎎/L IAA, 5.0㎎/L BA and 10.0㎎/L spermine. A Low frequency of regeneration occurred on hormone-free MS medium. Multiple shooks were regenerated at a pH of 4.0 to 8.0 on MS medium supplemented with 1.0 ㎎/L BA and 0.1 ㎎/L NAA. Polyamines promoted shoot- and root-formation by 2 to 4 times normal, Specific proteins associated with organogenesis were identified. Somatic embryogenesis occurred directly from the leaf blade, petiole and root segments cultured on MS medium with 2.0 ㎎/L BA and 2.0 ㎎/L BA and 2.0 ㎎/L NAA. Three types of regeneration in A, rusticana were clearly established, which could be applied to the study of morphogenesis and genetics at cell, tissue and organ levels.

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Medium Composition Affecting In Vitro Regeneration of Matteuccia struthiopteris (청나래고사리의 기내 포자체 재생에 미치는 배지 구성물질의 영향)

  • Shin, So Lim;Lee, Cheol Hee
    • FLOWER RESEARCH JOURNAL
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    • v.17 no.2
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    • pp.93-100
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    • 2009
  • This study was carried out to investigate the efficient in vitro mass propagation methods for juvenile sporophytes of Matteuccia struthiopteris. Chopped segments of pinnae, petiole and rhizome were cultured on 1/2MS with 0.1% activated charcoal. Among these explant sources only rhizome segments produced young sporophytes, regenerating vigorously on 1/2 MS medium. Adjusting sucrose concentration to 2% and supplement to $50mgL^{-1}$ $NaH_2PO_4$ in 1/2MS medium proved to be more efficient for plant regeneration. Various combinations of growth regulators such as kinetin, BA, NAA, and IBA were added to the growing media, and the best sporophyte regeneration was obtained by $1{\mu}M$ kinetin. The BA addition resulted in vigorous proliferation of meristematic tissues, but without differentiation to sporophytes. Three types of culture methods, solid using agar, liquid stationary, and liquid shaking culture, were employed with or without activated charcoal. The addition of 0.1% activated charcoal to modified 1/2MS media (2% sucrose, $50mgL^{-1}$ $NaH_2PO_4$, $1{\mu}M$ kinetin, pH 5.8 and 0.8% agar) yielded highest sporophyte regeneration in liquid shaking culture.