• Title/Summary/Keyword: Explants

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In vitro Propagation and Ex vitro Rooting of Tectona grandis (L.f ), APNBV-1 Clone

  • Ramesh, Kommalapati;Chandra, Mouli Kalla;Vijaya, Tartte
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.119-126
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    • 2009
  • An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.

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In vitro multiple shoot proliferation and plant regeneration in rose(Rosa hybrida L.)

  • Lee, Su-Young;Jung, Ji-Hye;Kim, Jeong-Hee;Han, Bong-Hee
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.223-228
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    • 2008
  • This study was conducted to investigate an optimal condition for shoot proliferation and regenerate shoots from in vitro leaflet and embryogenic calli from in vitro roots in rose. The effect of BAP on shoot proliferation was somewhat different depending upon genotypes or gelling agents. Leaflets with petiole cut from donor shoots which had been cultured in MS medium supplemented with 0.1 $mg{\cdot}L^{-1}$ NAA for six weeks was effective for regeneration of adventitious buds(ABs) as well as shoot elongation of Rosa hybrida cv. Sweet Pink. Culturing seven leaflet explants per petri plate($100mm{\times}15mm$) was effective for regeneration of ABs. Embryogenesis was shown in the calli induced from roots of Rosa hybrida cv. Sweet Pink cultured in the SH medium supplemented with 11 $mg{\cdot}L^{-1}$ 2, 4-D for four weeks. Color of calli induced from roots was yellow although their color was a little different as type of basal medium.

Shoot regeneration from internode sections of Ardisia pusilla DC.

  • Lee, Su-Young;Kim, Young-Soon;Han, Bong-Hee
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.209-213
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    • 2008
  • This study was conducted to regenerate shoots from internode sections(about 1mm in thickness) of Ardicia pusilla de Candolle. Internode sections were cultured on MS medium supplemented with TDZ or both TDZ and IBA. At one month after culture, primodium, which looks like protocorm like body(PLB) of orchid, appeared around swollen internodes. And then it grew and changed into the shape similar to granule of orange at two or three months after culture. At four to five months after culture, explants covered with them became a cluster, and then multiple shoots were regenerated from them. Primodia formation was the best when internode was cultured on MS medium supplemented with 0.25 $mg{\cdot}L^{-1}$ thidiazuron(TDZ) and 0.5 $mg{\cdot}L^{-1}$ indole-3-butyric acid(IBA). That internodes were cultured on MS medium supplemented with either higher concentration of TDZ than that of IBA, or equal concentration of TDZ and IBA, or TDZ only was little effective for primodia formation.

Analysis and Identification of Expressed Sequence Tags in Hairy Root Induced from Korean Ginseng (Panax ginseng C. A. Meyer)

  • Yang, Deok-Chun;In, Jun-Gyo
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.154-162
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    • 2004
  • Hairy roots were induced from Korean ginseng (Panax ginseng C. A. Meyer) root explants and studied for their gene expression. A total of 3,000 ESTs (expressed sequence tags) from ginseng hairy root were determined and about 2,700 ESTs have a length of readable sequence, which result in 1,352 unique ESTs sequences. The 879 ESTs showed significant similarities to known nucleotide or amino acid sequences in other plant species, which were divided into eleven categories depending upon gene function. The remaining 473 sequences showed no significant matches, which are likely to be transcripts or to be matched to other organisms. The results indicated that the analysis of the ginseng hairy root ESTs by partial sequencing of random cDNA clones may be an efficient approach to isolate genes that are functional in ginseng root in a large scale. Our extensive EST analysis of genes expressed in ginseng hairy root not only contributes to the understanding of the dynamics of genome expression patterns in root organ but also adds data to the repertoire of all genomic genes.

Factors Affecting on Shoot Formation in Bioreactor Culture of Rehmannia glutinosa Lib. (생물반응기(生物反應器)에서 지황(地黃)의 신초(新梢) 형성에 관여하는 요인(要因))

  • Park, Ju-Hyun;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.2
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    • pp.123-128
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    • 2000
  • This study was carried out to determine factors affecting on the mass propagation of Rehmannia glutinosa seedlings in bioreactor culture. Air-lift type bioreactor was more compatible to shoot formation than stirrer type. Fifty grams(90 stem explants) of inoculum in 1.5L medium was placed into 2.5L bioreactor with aeration rate of 0.5 v.v.m., which was proper for effective shoot formation. Adding MES as pH buffer to culture medium increased the numbers of shoot formation. Adding 5g/l of anti-vitrifying agent into culture medium was highly effective for diminishing the rate of vitrification in shoots formed.

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Transgenic Lettuce Expressing Chalcone Isomerase Gene of Chinese Cabbage Increased Levels of Flavonoids and Polyphenols

  • Han, Eun-Hyang;Lee, Ji-Sun;Lee, Jae-Woong;Chung, In-Sik;Lee, Youn-Hyung
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.467-473
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    • 2011
  • Flavonoid are large group of the polyphenolic compounds which are distinguished by an aromatic or phenolic ring structure and the phenolic compounds are induced by microbial infection, ultraviolet radiation, temperature and chemical stress. They are known for their antioxidant activity, anti-allergic, anti-inflammatory, anti-microbial and anti-cancer activities. In this study, changes in flavonoid content were investigated using heterologous chalcone isomerase (CHI) expression system. Also, phenolic compounds level was measured to examine the relation between flavonoids and phenols contents. Explants of lettuce (Lactuca sativa L.) were transformed with Agrobacterium tumefaciens LBA 4404 strain containing pFLH-CHI (derived from pPZP2Ha3) vector constructed with CHI gene from Brassica rapa. The putative transgenic plants were confirmed by genomic DNA PCR analysis. Also the transcription levels of the gene were analyzed by semi-quantitative RT-PCR with gene specific primers. The total flavonoid contents were increased at $T_0$ and $T_1$ generations over 1.4 and 4.0 fold, respectively. Total phenol contents also increased at $T_1$ generation. These results indicate that CHI gene plays an important role to regulate the accumulation of flavonoids and its component changes.

In Vitro Shoot Tip Culture of Haemaria discolor (Haemaria discolor 경정의 기내배양)

  • 왕영조;정재동;최수옥;지선옥
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.227-231
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    • 1994
  • The experiments were conducted to obtain culture conditions on optimal cultural conditions for propagation of a topical orchid, Haemaria discolor, through shoot tip cultrue in vitro. Survival percentage of shoot tip explant, in initial culture media was higher in H$_3$P$_4$ medium than in Murashige and Skoog's medium. Explants, cultured in H$_3$P$_4$ medium containing 1.0 mg/L kinetin, showed more shoot elongation when compared with those on other media. When the distal part of stem (pseudobulb) was longitudinally sectioned into half and cultured in H3P4 medium containing 0.1 mg/L 2ip, 0.1, or 1.0 mg/L kinetin, shoots from axillary bud were much more elongated under light condition.

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Advances in in vitro culture of the Brassicaceae crop plants

  • Park, Jong-In;Ahmed, Nasar Uddin;Kim, Hye-Ran;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.13-22
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    • 2012
  • Plant regeneration has been optimized increasingly by organogenesis and somatic embryogenesis using a range of explants with tissue culture improvements focusing on factors, such as the age of the explant, genotype, media supplements and $Agrobacterium$ co-cultivation. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in Brassicaceae crop plants. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in sexually incompatible species of $Brassica$. Crop improvement using somaclonal variation has also been achieved. Transformation technologies are being exploited routinely to elucidate the gene function and contribute to the development of novel enhanced crops. The $Agrobacterium$-mediated transformation is the most widely used approach for the introduction of transgenes into Brassicaceae, and $in$ $vitro$ regeneration is a key factor in developing an efficient transformation method in plants. Although many other Brassicaceae are used as model species for improving plant regeneration and transformation systems, this paper focuses on the recent technologies used to regenerate the most important Brassicaceae crop plants.

Isolation and identification of Agrobacterium spp. using carrot disc and transformation of Nicotiana gluca by selected strains (당근 절편을 이용한 Agrobacterium spp. 분리, 동정과 선발균주에 의한 연초의 형질전환)

  • Yang, Deok-Chun;Yang, D.C.;Choi, K.T.
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.1
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    • pp.43-52
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    • 1991
  • These studies were carred out to obtain the transformant from tobacco cells by Agrobacterium spp. from crown gall and soil at the natural field in Korea, and identified their virulence. Kodo's and Clark's selective media were used for isolation of Agrobacterium spp. In these media, total of 99 strains were characterized based on the morphological characteristics of colonies. Among them 34 strains were able to induce on carrot discs. And hypervirulent strains C23-1 and K29-1 were identified as Agrobacterium tumefaciens biotype 1 and biotype 2, respectively. These strains formed fast growing, larger gall as compared to those induced by other strains on the carrot discs. Transformed tobacco callus was initiated on the phytohormone free MS medium with 250$\mu\textrm{g}$/ml carbenicillin after co-cultivation of tobacco stem explants and Agrobacteria. On the phytohormone free media, shoot was rarely formed from transformed callus. However, these shoot were teratoma shoots which were not grown as normal shoot, and teratoma shoot from transformant by C23-1 was smaller than that of K29-1.

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Micropropagation of Hypericum erectum Thunberg by using Thidiazuron

  • Kim, Ok-Tae;Bang, Kyong-Hwan;In, Dong-Soo;Kim, Tae-Soo;Seong, Nak-Sul;Cha, Seon-Woo;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.5
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    • pp.278-281
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    • 2006
  • The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with $4.5\;{\mu}M$ TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was $10\;{\mu}M$. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.