• Title/Summary/Keyword: Plant tissue

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Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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Isolation and Characterization of Plant Pathogen that Cause Soft Rot Disease in Napa Cabbage (배추무름병 원인균 분리 및 특성 연구)

  • Kwon, Young-Hee;Yoo, Ah-Young;Yu, Jong-Earn;Kang, Ho-Young
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1177-1182
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    • 2009
  • In order to establish in vitro infection model for research of plant pathogen based on tissue softening disease in napa cabbage, eighty independent bacterial strains were isolated from the softened napa cabbage tissues. Eight bacterial isolates were primarily screened with the generation of reproducible tissue softening disease to fresh napa cabbages within 24${\sim}$48 hours after inoculation. Through various microbiological biochemical and morphological examinations, three Gram (-) isolates which harbor independent biological properties were finally chosen, and named as RBI, RB2 and RB6. Collective results obtained from API 20E test and analyses of VITEK 2 COMPACT and nucleotide sequences of 165 rRNA of each isolate proposed that isolates RBI and RB2 are close to the Erwinia carotovora subsp. odorifera, and RB6 is close to the Erwinia carotovora subsp. carotovora. These isolates grew optimally at $30^{\circ}C$ with neutral pH culture condition. The isolates caused softening tissue disease with dose-dependent manner regardless of pre-surface damages of napa cabbage. Minimum dose to cause soft rot disease for RBI, RB2 or RB6 were $8.0{\times}10^8$ CFU/mt $10^9$ CFU/ml or $4.7{\times}10^6$ CFU/ml respectively. These isolates caused tissue softening disease to eggplant, paprika and napa cabbage out of 14 different tested vegetables, indicating that these isolates damages specific plant tissues. The bacterial isolates obtained in this research and in vitro plant infection model will be adapted in the understanding of the mechanism of pathogenesis by plant pathogen.

Screening of Tissue Papers for Nematode Extraction for the Baermann Funnel Method (Baermann Funnel법의 선충 분리 여과지 선발)

  • Kang, Heonil;Eun, Geun;Ha, Jihye;Lee, Jaehyun;Kim, Donggeun;Kim, Yongchul;Choi, Insoo
    • Korean journal of applied entomology
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    • v.55 no.4
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    • pp.377-381
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    • 2016
  • The Baermann funnel method requires approximately four Kimwipe tissues for research a nematode count under a stereo microscope. To select more efficient and economical nematode extraction paper for nematode extraction, 15 different kinds of tissue papers were tested and compared with Kimwipe tissues. Nematode species used in the extraction efficiency tests include juvenile (J2) of Heterodera sp., J2 of Meloidogyne sp., Pratylenchus sp., Rhabditis sp., Acrobeloides sp., Panagrolaimus sp., Poikilolaimus sp. and Diplogasterida. The extraction efficiency varied between 42.0 to 88.8%. Considering costs, extraction efficacy, and clarity, the Pulling Kitchen Towel (Monalisa Co., Korea) is the best tissue, with clarity A, isolation efficiency of 69.4% (not significantly different compared to Kimwipe 1 ply 88.8%), and 1/2 costs per isolation of Kimwipe 2 ply.

In Vitro Tissue Culture Frequency and Transformation of Various Cultivars of Soybean (Glycine max (L.) Merr.) (다양한 콩 자원들의 기내 조직배양 효율 및 형질전환)

  • Seo, Mi-Suk;Cho, Chuloh;Jeong, Namhee;Sung, Soon-Kee;Choi, Man-Soo;Jin, Mina;Kim, Dool-Yi
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.278-286
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    • 2021
  • Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.

New Antisense RNA Systems Targeted Against Plant Pathogens

  • Matousek, J.;Vrba, L.;Kuchar, M.;Pavingerova, D.;Orctova, L.;Ptacek, J.;Schubert, J.;Steger, G.;Beier, H.;Riesner, D.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.379-385
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    • 2000
  • tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

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Genome-wide Identification, Classification, and Expression Analysis of the Receptor-Like Protein Family in Tomato

  • Kang, Won-Hee;Yeom, Seon-In
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.435-444
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    • 2018
  • Receptor-like proteins (RLPs) are involved in plant development and disease resistance. Only some of the RLPs in tomato (Solanum lycopersicum L.) have been functionally characterized though 176 genes encoding RLPs, which have been identified in the tomato genome. To further understand the role of RLPs in tomato, we performed genome-guided classification and transcriptome analysis of these genes. Phylogenic comparisons revealed that the tomato RLP members could be divided into eight subgroups and that the genes evolved independently compared to similar genes in Arabidopsis. Based on location and physical clustering analyses, we conclude that tomato RLPs likely expanded primarily through tandem duplication events. According to tissue specific RNA-seq data, 71 RLPs were expressed in at least one of the following tissues: root, leaf, bud, flower, or fruit. Several genes had expression patterns that were tissue specific. In addition, tomato RLP expression profiles after infection with different pathogens showed distinguish gene regulations according to disease induction and resistance response as well as infection by bacteria and virus. Notably, Some RLPs were highly and/or unique expressed in susceptible tomato to pathogen, suggesting that the RLP could be involved in disease response, possibly as a host-susceptibility factor. Our study could provide an important clues for further investigations into the function of tomato RLPs involved in developmental and response to pathogens.

Production of Corydalis Alkaloids by Plant Cell Culture(I) (식물세포배양에 의한 Corydalis Alkaloid의 생산(I))

  • Chang, Jung-In;Shin, Seung-Won;Chi, Hyung-Joon
    • Korean Journal of Pharmacognosy
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    • v.26 no.4
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    • pp.419-425
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    • 1995
  • Corydalis remota Fish. ex Max. (Papaveraceae) is a well known medicinal plant being used as analgesics or anticonvulsive in oriental medicine. As the alkaloid content is known to vary depending on the environmental factors, the technology of plant tissue culture can be adopted as source of Corydalis-alkaloids. The present study describes an establishment of tissue cultures of Corydalis which produce alkaloids consistently. Callus were induced from immature seeds of Corydalis remota by placing the seeds on MS static media containing NAA(0.25, 1.0 and 4.0 mg/l, respectively). The combined treatment of NAA(1.0 mg/l) with cytokinin(BAP 0.5 mg/l) improved the induction of callus. TLC scanning data followed by sequential extraction and purification revealed that the induced callus contains a significant amount of alkaloids. Cell suspension cultures were established by transferring the induced callus into the liquid media with the same condition of plant growth regulators as the callus culture.

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Effect of Plant Growth Regulators on Calls Initiation and Organogenesis from Tissue Culture of Arabidopsis thaliana Stem (애기장대 줄기 조직배양에 있어서 식물생장조절제가 캘러스 형성과 기관분화에 미치는 영향)

  • Park, Jung-An;Park, Jong-Bum
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.257-261
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    • 2003
  • This experiment was carried out to investigate the effects of plant growth regulators on the organogenesis from the tissue culture of Arabidopsis thaliana stem, and the origin of the callus development. When the stem segments were cultured on medium with 2mg/L IAA or NAA, adventitious roots and trichomes were differentiated after 11 days of culture. Callus vigorously formed on medium with 2/L2,4 after 7 days of culture, but adventitious roots and trichomes were not differentiated from callus after 10 days of culture. This results suggesting that picloram is very effective auxin for the callus formation and organogenesis. Callus weakly formed on 0.05mg/L kinetin, and formed on combination of auxins(2mg/L) with 0.05mg/L kinetin. But the effect of combination of auxins and kinetin the callus formation was less than 2,4-D or picloram alone. A histological examination of callus formed on picloram showed that phloram showed that phloem parenchyma cells were divided and enlarged after 2 days of culture. Cortex parenchyma cells were divided and meristematic nodules were developed from these cells after 4 days of culture. Finally, callus formed on outside of cortex and epidermis by division of meristematic nodules after 7 days of culture.

Cyclized Induction of Phenylalanine Ammonia-Lyase Gene Expression in Rhizoctonia solani-Infected Stems of Tomato

  • Yeo, Yun-Soo;Kim, Soo-Jin;Koo, Bon-Sung;Lee, Churl-Ho;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.151-156
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    • 2004
  • Soil-borne fungal pathogens such as Verticillium and Rhizoctonia can colonize in the stem tissue of plant through root and lead to wilting symptoms of plant by blocking. water transportation. During the colonization of Rhizoctonia solani in the vascular tissue of tomato stems, particularly, phenylalanine ammonia-lyase (PAL) gene induction pattern was cyclized showing peak induction at two different time points (10 and 80 h) after fungal spores inoculation in vivo. In leaves or roots, however, no such cycling pattern was observed. The first induction peak may be due to an initial sporulation events leading to a second induction peak by a proliferation of fungal spores to the upper stems or other tissues from an initial spore trapping sites. Tomato PAL gene was also dramatically induced by wounding, light illumination and mercury chloride treatment but was not cyclized. Mercury chloride showed the earliest induction with all tissues even at half an hour after treatment.

GUS Expression by CaMV 35S and Rice Act1 Promoters in Transgenic Rice

  • Kwang-Woong Lee
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.371-380
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    • 1994
  • To determine the patterns and the levels of expression of the cauliflower mosaic virus (CaMV 35S) promoter and the rice actin 1 (Act1) promoter in rice, transgenic rice plants containing CaMV 35S-$\beta$-glucuronidase (GUS) and Act1-GUS constructs were generated and examined by fluorometric and histochemical analyses. The fluorometric analysis of stably transformed calluses showed that the activity of the rice Act1 promoter was stronger than that of the CaMV 35S promoter in rice cells. In a histochemcial study of the transgenic rices, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in parenchymal cells of vascular tissues of leaves and roots and mesophyll cells of leaves. These results are similar to those of potato, a dicot plant. In contrast, rice plant transformed with Act1-GUS fusion construct revealed strong GUS activity in parenchymal cells of vascular tissue, mesophyll cells, epidermal cells, bulliform cells, guard subsidiary cells of leaves and most cells of the root, suggesting that the rice Act1 promoter is more constitutive than the CaMV 35S promoter. It was also confirmed that in both types of transgenic rice little or no staining was localized in metaxylen tracheary elements of vascular tissue from leaves or roots. These results indicate that the rice Act1 promoter can be utilized more successfully for expression of a variety of foreign gene in rice than the CaMV 35S promoter.

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