• Title/Summary/Keyword: Explants

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Transfer and Expression of a Tomato Inhibitor II and $\beta$-Glucuronidae Fusion Gene in Flowering Cabbage, Brassica oleracea var. acephala DC. (꽃양배추로의 프로타제 저해제 II와 $\beta$-Glucuronidase 융합 유전자의 도입 및 발현)

  • 김창길;정재동;안진흥;김경민
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.37-43
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    • 1998
  • The highest transformation frequency was observed when cotyledonary and hypocotyl explants of flowering cabbage (Brassica oleracea var. acephala DC) 'Eunbae' were cultured on shoot induction medium without kanamycin for 1 day, then cocultured with Agrobacterium tumefaciens LBA4404;;pGA1036 harboring tomato inhibitor II promoter and $\beta$-glucuronidae (GUS) fusion gene for 3 days. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. Incorporation of the GUS gene into flowering cabbage was confirmed by PCR analysis of DNA. Southern blot analysis showed that ECL-labeled GUS gene was hybridized to the expected amplified genomic DNA fragment of about 366 bp from transgenic flowering cabbage. Histochemical analysis based on the enzymatic activity of the GUS protein indicated that PI-II promoter activity was sysmatically associated with vascular tissue in wonded as well as in non-wounded leaves, petioles and stems, but not in roots. Partial wounding with razor blade showed not systemic induction but partial induction.

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Studies on the Induction of Transformation in Cereal Plane V. Transformation of Wheat by Electroporation (곡물류의 형질전환 유도에 관한 연구 V. Electroporation에 의한 밀의 형질전환)

  • Song, Jung-Won;Jung, Byung-Kyun;Bae, Dong-Kyu;Im, Hyung-Tak;Nam, Back-Hee;Jung, Hyun-Sook;Hwang, Baek
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.187-192
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    • 1994
  • Wheat (Triticum aestium L.cv Cho-Kwang) explants were transformed by electrporation. Excised root segments were elechoporated with plasmid DNA of pBI121 and transferred to medium containing 300 mg/L kanamycin. Transformed calli formed within 5-7 days of culture and were selected from electroporated tissue on medium containing kanamycin after 4 weeks. The highest transformation frequency was obtained after electroporation with a pulse of 200 V/800 uF and calli formed at frequencies up to 2.5%. GUS ($\beta$-glucuronidase) assay and dot blot analysis showed that the foreign gene was capable of expressing in root explants subjected to electroporation and calli derived from the explants..

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Effect of Genotype and Explant on Somatic Embryogenesis and Acclimatization of Acanthopanax senticosus (가시오갈피의 수집종과 배양조직에 따른 체세포배발생 및 재분화 식물체의 순화)

  • Lee, Cheng-Hao;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.3
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    • pp.217-221
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    • 2002
  • Callus induction and embryogenesis were studied in three different genotypes of Acanthopanax senticosus, to develop a protocol for somatic embryogenesis and acclimatization. Young leaf, stem, node, petiole, peduncle, flower and root explants were collected from 3-year old trees of A. senticosus accessions (Korea, Russia and Japan). Callus was obtained from all cultured explants but showed the higher rate of callus formation in flower cultured. For the three A. senticosus accessions, callus was well formd on MS media containing 2mg/ l of 2,4-D and 2mg/ l of TDZ, 4mg/ l of 2,4-D and 1mg/ l of TDZ than other treatments. For three A. senticosus accessions, when callus transferred to MS medium with 2,4-D, embryogenic cell formed. For A. senticosus accessions Korea, embryogenic cells were obtained on callus induced from petiole, stem, node and root explants, and induction rate was lower than 3%. 200mg of embryogenic callus was transferred to MS free liquid medium and somatic embryos of heart stage were obtained after 45days of culture. When somatic embryo of germination stage were transferred to solid medium, most of the embryos were regenerated into plantlets on 1/4 MS medium. Normal plants with both shoots and roots were transferred to greenhouse soil and were successfully acclimatized.

High Frequency Shoot Formation and Plant Regeneration from Cotyledonary Hypocotyl Explants of Boxthorn (Lycium chinense Mill.) Seedlings (구기자나무(Lycium chinense Mill.) 유식물체의 자엽하배축으로부터 고효율의 부정아 형성과 식물체 재생)

  • Jo, Man-Hyun;Ham, In-Ki;Lee, Bong-Chun;Kim, Jae-Whune;Lee, Won-Seok;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.203-207
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    • 2004
  • To establish high frequency shoot formation from two cultivars (Cheongyangjaerae and Myungan) of boxthorn (Lycium chinense Mill.), hypocotyl segments with cotyledons from seedlings were used as explants. High frequency adventitious shoot formation (more than 80%) were obtained from hypocotyl segments with cotyledon on MS medium supplemented with 1.0 mg/L zeatin, when precultured for 3 weeks under dark conditions followed by transfer to light conditions. But there was no shoot induction in the explants cultured without preculture under dark conditions. Roots were induced from the shoots when transferred to rooting medium supplemented with 1.0 mg/L IAA for 4 weeks. Regenerated plantlets were grown to normal mature plants in soil.

Interaction of Bovine Growth Hormone with Buffalo Adipose Tissue and Identification of Signaling Molecules in Its Action

  • Sodhi, R.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1030-1038
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    • 2007
  • Results on localization of growth hormone receptor (GHR), interaction of growth hormone (GH) with receptor in buffalo adipose tissue and identification of activated signaling molecules in the action of GH are presented. Bovine GH (bGH) was labeled with fluorescein or biotin. Fluorescein-labelled bGH was used for localization of GHRs in buffalo adipocytes. The receptors were present on the cell surface. The affinity of binding of GH to its receptor was determined by designing an experiment in which buffalo adipose tissue explants, biotinylated GH and streptavidin-peroxidase conjugate were employed. The affinity constant was calculated to be $2{\times}10^8M^{-1}$. The receptor density on adipose tissue was found to be 1 femto mole per mg of tissue. Signalling molecules generated in the action of GH were tentatively identified by employing Western blot and enhanced chemiluminescence techniques using anti-phosphotyrosine antibody. Based on molecular weights of proteins reactive to anti-phosphotyrosine antibody, three signaling molecules viz. insulin receptor substrate, Janus activated kinase (Jak) and mitogen activated protein were tentatively identified. These signaling molecules appeared in a time (incubation time of explants with growth hormone) dependent way. The activation of Jak2 was confirmed by employing anti-Jak2 antibody in a Western blot. The activation of Jak2 occurred during 5 min incubation of buffalo adipose tissue explants with GH and incubation for an additional period, viz. 30 min. or 60 min., resulted in a drastic reduction in activation. The results suggest that Jak2 activation is an early event in the action of GH in buffalo adipose tissue.

In vitro shoot initiation of Artocarpus heterophyllus Lam. (Jak Fruit) Effect of the explant type and the season of explant collection

  • Kahk, Kasturiarachchi;Wtpsk, Senarath;Lee, Kui-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10b
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    • pp.2-3
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    • 2003
  • A method for rapid propagation of mature Jack fruit was developed. Four types of explants (mature embryos, apical meristems of young seedlings, apices from mature plants and nodal segments) were used. It has been found 88% of young apical meristems produced shoots in Campbell and Durzan (CD) medium compared to 60% in Murashige and Skoog (MS) medium. Only 1/3 of them produced multiple shoots. Shoot initiation from nodal segments was very rare. Mature apices produced callus. Although removal of the sheathing cover around mature buds enhanced the shoot initiation but success rate was low in growth regulator free medium. Embryos respond to the CD medium but not to the MS medium. Embryos from seeds soaked in water for 24 hours produced shoots after 8 weeks of incubation and the success rate was 70% while embryos from dry seeds only produced roots. There was no significant effect of cold storage (refrigeration) for 7 days on shoot initiation from mature embryos (65%) but the ability for shoot induction declines with storage time (55% after 21 days of cold storage). Mature axillary buds were established in Modified Campbell and Durzan (CD) medium supplemented with 0.5mg/1 and IBA. There was a significant difference in the growth performance of shoots according to the period of the year in which explants were collected. Highest (60%) was observed in November-January period. It was only 30% when the explants were collected in February-April or May-July and decreased to 20% in August-October. The shoots produced in November-January showed a higher vigor than those produced in other months. Since Jak fruit show seasonal changes in fruit bearing and shedding of leaves, it can be suggested that the difference in growth performances of tissues cultured in artificial culture media would have been affected by endogenous rhythms.

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Measurement of Determination Time of In-Vitro Flowering in Ginseng (Panax ginseng) (인삼의 기내 개화 결정시기의 측정)

  • 이행순;이광웅;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.6
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    • pp.347-351
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    • 1994
  • To measure the time required for ginseng explants to become determined to form flower buds, we cultured zygotic embryos, seedlings, and cotyledonary nodes on MS medium supplemented with BA and GA$_3$of 5 ${\mu}$M each (flower inducing medium, FIM) for various periods and transferred to the basal medium. The explants required a minimum of 10 days on FIM to be determined. Histological observations revealed that the axillary meristem to be fated to develop into flower bud remained in a state of shoot meristem during the first 10 days of culture and differentiated into flower bud after 15 days of culture. We suggest that the in-vitro flowering system described in this study is useful in investigating (a) regulatory element(s) to cause the phase change from the vegetative to reproductive state by comparing predetermined explants with determined ones at the molecular level.

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Plantlet Formation and Somatic Embryogenesis from the penduncle explants of wild Corydalis remota for. peatinata (야생 현호색 (Corydalis remota for. peatinata)의 꽃자루로부터 체세포배 발생과 소식물체형성)

  • 천우재;이대원
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.15-19
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    • 1999
  • For the induction of somatic embryogenic callus, the penduncle explants of Corydalis remota for peatinata were cultured on MS basal media supplemented with 2,4-D, kinetin and zeatin. The highest embryogenic callus formation was observed on the media containing 2.0 mg/L of 2,4-D and 2.0 mg/L of zeatin. The somatic embryogenesis on the media with 0.5 mg/L of cytokinin (zeatin or kinetin) were excellent under light condition, however somatic embryos abnormally developed into plantlets. Normal dicotyledonary plantlets were found on MS medium supplemented with 1.0 mg/L of zeatin. When MS medium with 2,4-D plus cytokinin and with BAP were used, the secondary somatic embryogenesis took place in root explants of the regenerants derived from in vitro somatic embryogenic callus.

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High Frequency Adventitious Shoot Formation and Plant Regeneration in Leaf Explant Cultures of Ixeris sonchifolia Hance, a Newly Proposed Model Plant for Organogenesis

  • Min Sung-Ran;Kim Young-Hoe;Jeong Won-Joong;Han Su-Kyung;In Don-Su;Liu Jang R.
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.221-224
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    • 2003
  • Leaf explants of Ixeris sonchifolia produced adventitious shoots at a frequency of 100% when cultured on MS medium supplemented with combinations of various concentrations of 6-benzyladenine (BA) (0.44, 4.44, or 8.87 ${\mu}M$) and 0.54 ${\mu}M$ NAA, or MS medium supplemented with 22.19 ${\mu}M$ BA and 2.69 ${\mu}M\;\alpha$-naphthaleneacetic acid (NAA) after four weeks of culture. Each explants (approximately $3{\times}6mm$) produced greater than 70 shoots at a combination of 0.44 ${\mu}M$ BA and 0.54 ${\mu}M$ NAA. Leaf explants produced shoots at a frequency of greater than 80% even at as low as 0.13 ${\mu}M$ BA as the sole growth regulator. Upon transfer to one-third strength MS with 0.54 ${\mu}M$ NAA, excised adventitious shoots were rooted at a frequency of 100%. Regenerated plantlets were transplanted to potting soil and grown to maturity in a greenhouse. The competence of I. sonchifolia for plant regeneration via organogenesis appears to be greater than the competence of tobacco, currently the best model plant for organogenesis.

In vitro introduction adventitious shoots and plant regeneration of sengon (Paraserianthes falcataria (L.) Nielsen) (셍온(Paraserianthes falcataria (L.) Nielsen)의 기내 부정줄기 유도 및 식물체 재분화)

  • Kim, Ji Ah;Moon, Heung Kyu;Kim, Yong Wook;Bae, Eun Kyung
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.235-238
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    • 2015
  • Adventitious buds were obtained from isolated cotyledons cultured on MS medium with various concentrations of 6-benzylamino purine (BA) and thidiazuron (TDZ). The highest numbers of adventitious buds were obtained on MS medium supplemented with 0.2 mg/L BA. Experimental culturing with half the petiole portion and half with the terminal segments were grown on MS medium contained with 0.2 mg/L BA. Frequency of the adventitious bud induction was variable accordingly to the type of cultured explants. Explants with the half petiole showed the highest adventitious bud induction rate (80%) compared to explants of half with terminal segment (20%). An elongated shoot from the buds and growth of advent roots were both possible on the 1/2 MS medium without a plant growth regulator. These results offer an effective way in which clonal propagation can be accomplished.