• Title/Summary/Keyword: plant tissue culture

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Analysis of Tissue Plasminogen Activator Expression using Pollen Culture in vitro (기내 화분배양을 이용한 Tissue Plasminogen Activator 발현분석)

  • 박인혜;박희성
    • KSBB Journal
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    • v.17 no.6
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    • pp.582-585
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    • 2002
  • In an effort to use plant biotechnology for the production of biopharmaceuticals, pollens collected from lily (Lilium longiflorum) were grown in vitro and transformed with a PCR-amplified 1.7 kb cDNA encoding human tissue plasminogen activator (tPA) using Agrobacterium via a vacuum infiltration process. Western blotting showed that transgenic lily pollen tubes selected on kanamycin for 16 hrs expressed a tPA protein with a size similar to the human standard, suggesting their possible use as a disposable host for rapid foreign protein production.

Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Effects of Inorganic Nitrogen released from Roots on the Nitrogen Metabolism (뿌리 방출물중 무기태질소가 체내성분 변이에 미치는 영향)

  • 소상섭
    • Journal of Plant Biology
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    • v.22 no.1_2
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    • pp.5-14
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    • 1979
  • In several leguminous plants such as acasia, arrowroot and bushclover, growth rate and contents of nitrogen, phosphorus and potassium in the tissues and the variation in the culture media were determined. In water cultrue which was free of added nutrients, nitrogen was found to be largely in the form of nitrate(NO3-N). This NO3-N is believed to be the result of nitrification from NH4-N which was apparently released form the plants. From the studies of organ culture with root segments, the amount of nitrogen released and absorbed was found to be proportional to the amount added to the mediuim. Especially, in the N-plot, the amount of nitrogen absorbed by the tissue reached more than 90% of the amount supplied to the medium already in early stage. On the contrary, in the amount free plot, the amount of nitrogen released from the tissue was lower than the minimum level in the N-plot. The amount of total N and P in the cultured tissue was found to be influenced by the amount of nitrogen addedin the medium. However, the amount of K in the tissue was not related to the nitrogen level in the medium, but rather it was influenced by the amount of added potassium. These findings present little difference in the metabolic pattern among the three species plants studied, and suggest that the woody leguminous plants have some common features in tehir metabolic pattern.

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Induction of in vitro root tubers in Holostemma annulare (Roxb.) K. Schum. for the production of bioactive metabolites

  • Smitha Devi, Padmavathi Amma Somasekharan Nair;Hemanthakumar, Achuthan Sudarsanan;Preetha, Thankappan Suvarna
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.230-239
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    • 2022
  • Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

Effects of plant preservative mixtureTM on in vitro germination of Dendrobium thyrsiflorum Rchb.f. and its application in orchid conservation

  • Tran Trung Chanh;Nguyen Tan Huy;Nguyen Thu Ha;Khanh Le;Nguyen Huu Hoang
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.108-114
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    • 2023
  • In vitro conservation is one of the most effective strategies for rare plant protection, especially for orchid species. To maximize the success rates of in vitro explant establishment (stage I) in conservation programs, the application of tissue culture additives such as Plant Preservative MixtureTM (PPMTM) should be emphasized. In this study, we used Dendrobium thyrsiflorum Rchb.f. (1875) seeds and seedlings as a model for the evaluation of PPMTM's phytotoxicity in the meristematic tissues of epiphytic orchids. PPMTM had no observable inhibitory effect on protocorm, shoot, or root development when it was supplemented at 0.1%. PPMTM supplementation caused adverse effects on D. thyrsiflorum explants at concentrations > 0.2%. At high concentrations, young in vitro seedlings showed damage, especially at the root tissue level. Based on this model, supplementation of 0.1-0.2% PPMTM to culture media was successfully implemented to establish in vitro cultures of other rare orchid species in our conservation program.

Review on the development of virus resistant plants in Alstroemeria

  • Park, Tae-Ho;Han, In-Song;Kim, Jong-Bo
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.370-378
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    • 2010
  • This review describes the stratagies of development of virus-resistant Alstroemeria plants using the genetic modification system. Despite of increasing of its importance in cut flower market, improvements of some horticultuirally important traits such as fragrance, long vase-life, virus resistance and tolerance against abiotic stresses are lack of the breeding program in Alstroemeria. Of these traits, virus-resistance is quite difficult to develop in Alstroemeria plants due to the limitations of genetic variation in the existed germplasm. To extend the genetic variation, plant biotechnological techniques such as genetic transformation and tissue culture should be combined to develop virus-resistant line in Alstroemeria. In this review, several strategies for the generation of virus-resistance by using natural resistance genes, pathogen-derived genes and other sources including pathogen-derived proteins, virus-specific antibodies and ribosome-inactivating proteins are presented. Also, brief histories of breeding, tissue culture, and transformation system in Alstroemeria plants are described to inderstand of the application of transgenic approach for the development of virus-resistance in Alstroemeria species.

Enhancing in vitro Growth of Bulbs for Mass Propagation of Lily Germplasm

  • Song, Jae-young;Lee, Young-yi;Yi, Jung-yoon;Lee, Jung-ro;Yoon, Mun-sup
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.17-22
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    • 2021
  • Plants regenerated from in vitro cultures carry chromosomal variations, especially in long-term culture. Reducing the duration of plant tissue culture is one of the ways to reduce genetic and epigenetic changes. In this study, we reduced the duration of long-term culture and repeat subculture using small bulblets derived from bulb scales in two lily cultivars. The adventitious bulblets derived from bulb-scale tissue were cultured on three different media containing Murashige and Skoog (MS) basal medium supplemented with 1 g/L Charcoal, MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone with or without Charcoal, respectively. About seven weeks later, the number of newly propagated multiple shoots in the two media, A and B media, showed little differentiation. Compared to both media, the number of propagated multiple shoots increased 5-fold in MS medium containing 0.3 mg/L IAA and 0.4 mg/L BA hormone without Charcoal (C medium). The number of propagated multiple shoots ranged from 5 to 6 and 4 to 6 with an average of 5 in TropicalPink and GreenStar cultivars, respectively. The flow cytometric measurements indicated no variation in the ploidy level between control and in vitro propagated plants.

Effect of tissue proliferation and somatic embryo induction in Larix kaempferi following treatment with organic nitrogen sources and plant growth regulators (일본잎갈나무(Larix kaempferi) 유기질소원 및 식물생장조절물질 처리에 따른 조직증식 및 체세포배 유도 효과)

  • Kim, Yong Wook;Kim, Ji Ah;Moon, Heung Kyu;Jeong, Su Jin
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.376-379
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    • 2015
  • This study was conducted to evaluate the effects of different types and concentrations of organic nitrogen sources (${\small{L}}$-Glutamine and casein hydrolysate, CH) and plant growth regulators (auxins and cytokinins) on embryogenic tissue proliferation and somatic embryo production in L. kaempferi. Overall, the highest tissue fresh weight was obtained at either 2 or 4 weeks in culture when 1,000 mg/L ${\small{L}}$-Glutamine was added to the culture medium, which showed similar results with other treatments. In experiments with different types and concentrations of plant growth regulators on somatic embryo production, the highest production (426.3/90 mg tissue) was found when 0.2 mg/L IBA was added; however, no somatic embryos were induced following treatment with 0.2 mg/L BA or Kinetin. The effect of various concentrations of IBA on somatic embryo production was also tested. The best result (303/90 mg tissue) was obtained when plants were treated with 0.2 mg/L IBA; 1.0 mg/L IBA was also effective (281/90 mg tissue). The lowest result (109.3/90 mg tissue) was obtained with 5.0 mg/L IBA.

Purification and Partial Characterization of a Peroxidase from Perilla Callus

  • Hur, Yeon-Jae;Lee, Han-Gil;Hu, Gaosheng;Chung, Won-Bok;Jeong, Soon-Jae;Yi, Young-Byong;Nam, Jae-Sung;Chung, Young-Soo;Lee, Jai-Heon;Kim, Doh-Hoon
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.355-361
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    • 2007
  • Cotyledons of perilla6 were cultured on MS medium containing 0.5 mg/l NAA and 0.5 mg/l BA for 7 weeks. The activity of perilla peroxidase was observed to increase following culture stages as assessed by peroxidase assay. A peroxidase (POD) was purified from perilla tissue cultured on MS medium for 7 weeks. The peroxidase was purified using ion exchange and gel nitration chromatography. The perilla peroxidase had a molecular mass of 30 kDa by SDS-PAGE. We showed that the N-terminal amino acid sequence of this protein shared 67% identity with the tea peroxidase. As indicated by SDS-PAGE, the banding pattern of the 30 kDa polypeptide present in total soluble protein from perilla tissue was increased following culture stages. Immunoblot analysis indicated that perilla peroxidase protein appeared after 3 weeks of perilla tissue culture, and continued to increase with extended duration of tissue culture for at least 7 weeks.

Current status on applications of conventional breeding techniques and biotechnological system in ornamentals

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.107-117
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    • 2020
  • Flower industry is now growing due to the development of economy in many countries. Simultaneously, needs from consumers in flower market are varied widely. To satisfy the needs from consumers and deal with a variety of diseases from a lots of pathogens as well as climate change, new elite flower cultivars should be released in flower market. For this purpose, conventional and biotechnological techniques can be employed to make good cultivar. Therefore, this review describes the general overview of flower breeding techniques including cross-hybridization, mutation breeding and genetic transformation systems. Also, breeding systems for ornamentals derived from plant tissue culture such as embryo culture, in vitro fertilization, ovary/ovule culture and haploid production were reviewed. Furthermore, in this study recent development of the generation of new flower cultivars using marker-assisted breeding, plant transformation including particle bombardment and Agrobacterium tumefaciens as well as genome-editing technology were described. This review will be contributed to the development and releasement of new flower cultivars with horticulturally useful traits in the future.