• Horticultural Science & Technology
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• v.33 no.1
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• pp.124-132
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• 2015

Echinacea purpurea (L.) Moench (purple cone flower) is an important medicinal plant; it can enhance immunity, relieve pain, and reduce inflammation, and also has hormonal, antiviral, and antioxidant effects. Adventitious root biomass of Echinacea purpurea was produced in commercial-scale bioreactors for use as a dietary supplement in the food industry and in traditional medicine. Biosafety and toxicological evaluations of tissue-cultured Echinacea purpurea adventitious roots (TCEPARs) were performed. Reverse mutation and chromosomal aberration tests showed no significant mutagenicity. Furthermore, repeated four-week oral dose tests performed in Sprague-Dawley rats did not show any notable changes in the general behavior of the rats, in the gross appearance of their internal organs, or in their mortality rate. There were no differences between the control group and the treatment group in parameters such as absolute body weight, hematology, blood chemistry, and absolute and relative organ weights. These findings indicate that TCEPARs are safe and nontoxic when consumed at an average dietary level and can be used as raw material for traditional medicine and the food industry.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.10a
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• pp.50-58
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• 2003

Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase + 0.4% macerozyme, 1% cellulase + 0.2% macerozyme and 0.5% cellulase + 0.1 % macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5 ${\times}$ 108protoplasts/ml/ g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

• Journal of Forest and Environmental Science
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• v.35 no.1
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• pp.54-60
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• 2019

Plant tissue culture techniques offer quick methods of regeneration of plants of medicinal importance but the survival chances of such plants are always questionable when shifted to the in vivo conditions. The present study enumerates the micromorphological developments in the leaves of in vitro regenerated and field transferred plantlets of Oldenlandia umbellata. The leaves developed in vitro after $4^{th}$ subcultures of multiplication phase and after 6 weeks of field transferred plants were used. Statistically significant differences in the number of stomata, veins, raphides, crystals and trichome density per square mm were observed. The improvements in stomatal apparatus and density (decreased from 41.85 to 32.20), developments in leaf architectural parameters and emergence of defense mechanism through increased numbers of raphides (8 to 15), crystals and trichomes (13.5 to 18.2) proved acclimation of tissue culture raised plantlets from in vitro to the in vivo environments lead to 100 % success in field establishment of the plantlets. The in vitro induced foliar abnormalities (changes in stomata, venation pattern, vein density, trichomes, crystals etc.) were repaired while hardening of plantlets in the greenhouse and finally in the field. The observed micromorphological response of leaves under altered environmental conditions could help in determination of proper stage of field transfer and prediction of survival percentage of in vitro regenerated O. umbellata plantlets.

• Korean Journal of Veterinary Service
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• v.42 no.1
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• pp.39-42
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• 2019

Tetanus is an acute, often fatal, and infectious disease of all species of domestic animals caused by the neurotoxin of Clostridium tetani (C. tetani). This disease is usually known to develop after microbial contamination in the deep or penetrating wound sites. In February 2017, a farmer who was raising 76 cows injected foot and mouth disease vaccine to three or more cows with one syringe. Their clinical symptoms were observed 2 to 16 days after the vaccination. The initial symptoms were stiffness, rigidity of the neck and limbs, pricked ears, and prolapse of the third eyelid. Subsequently, there was recumbency with extension of the limbs, convulsions and opistotonus and the affected 20 cows were all died. Two dead cows were submitted to Animal and Plant Quarantine Agency for disease diagnosis. At necropsy, a focal edematous abscess of 15 to 20 cm in diameter was grossly observed in the subcutaneous and intramuscular tissue of scapular region and filled with a large amount of greenish pus. The feed was full in oral cavity and slightly observed in the trachea and lobes of lung. Histopathologically, focal granulomatous nodules with eosinophilic materials in the tissue were observed. In the lung, aspiration pneumonia and gram (+) bacteria were seen. The C. tetani was isolated in samples anaerobically cultured using reinforced clostridial medium and identified by PCR. To our knowledge, no previous outbreak of tetanus in cattle has affected such a high number of animals; neither has it been associated with misuse of the same syringe and needle to administer multiple individuals.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2004.10a
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• pp.65-67
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• 2004

During initial interactions of bacteria with their host plants, most plants recognize the bacterial infections and repel the pathogen by plant defense mechanism. The most active plant defense mechanism is the hypersensitive response (HR) which is the localized induced cell death in the plant at the site of infection by a pathogen. A primary locus induced in gram-negative phytopathogenic bacteria during this initial interaction is the Hrp locus. The Hrp locus is composed of a cluster of genes that encodes the bacteral Type 111 machinery that is involved in the secretion and translocation of effector proteins to the plant cell. DNA sequence analysis of hrp gene in phytopathogenic bacteria has revealed a Hrp pathogenicity is]and (PAI) with a tripartite mosaic structure. For many gram-negative pathogenic bacteria, colonization of the host's tissue depends on the type III protein secretion system (TTSS) which secrets and translocates effector proteins into the host cell. Effectors can be divided into several groups including broad host range effectors, host specific effectors, disease specific effectors, and effectors inhibit host defenses. The role of effectors carrying LRR domain in plant resistance is very elusive since most known plant resistance gene carry LRR domain. Host specific effectors such as several avr gene products are involved in the determination of the host specificity. Almost all the phytopathogenic Xanthomonas spp. carry avrBs1, avrBs2, and avrBs3 homologs. Some strains of X. oryzae pv. oryzae carry more than 10 copies of avrBs3 homologs. However, the functions of all those avr genes in host specificity are not characterized well.;

• Current Research on Agriculture and Life Sciences
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• v.11
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• pp.11-17
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• 1993

The transfer of genetic material into pea tissue was accomplished by using an avirulent strain of Agrobacterium tumefaciens containing the binary vector. The method used for transformation requires non-tissue culture steps as it involves the inoculation of the site of the shoot removed of germinating seeds. The identification of ${\beta}$-glucuronidase activity in the tissues of $T_0$ pea plants indicates that the plant expressible ${\beta}$-glucuronidase gene, contained the T-DNA region from pLPBO2, had been transferred at least into somatic tissues. Putative transformed $T_0$ pea plants were advanced to produce $T_1$ plants which were also assayed for the presence of the transferred ${\beta}$-glucuronidase gene. The presence of the ${\beta}$-glucuronidase gene in DNAs isolated from $T_1$ plant was demonstrated by DNA gel blot hybridization. This analysis revealed that the transformed plants contained ${\beta}$-glucuronidase gene.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.41 no.6
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• pp.704-709
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• 1996

This experiment was carried out to establish the system of mass propagation through tissue culture using sucker and stem in Ramie. The sterilization for tissue culture of Ramie was the better treatment of 2% NaClO for 20 minute into ultrasonic cleaner than the others, and so rate of contamination was 3.3%, and it was able to produce 96% healthy plant. The effect of growth regulator was superior to mixed treatment of 0.02mg/$\ell$ NAA, 1.5mg/$\ell$ BA, 0.lmgmg/$\ell$ GA$_3$, which it was not formed callus and but produced 96% healthy plant. The effect of propagation was higher in culturing of the stem node than the sucker in cultural part, local variety than improved ones. The effect of acclimatization was superior to pretreatment of 30 minute after soaking in 100ppm NAA, transplanting on bed soil which mixed to ratio of vermiculite : soil : sand =1 : 2 : 1, the transplanted plants were grown all normal.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.257-263
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• 2001

Species of the genus Orobanche are parasitic flowering plants, holoparasites, which cling to the roots of green plants. Their tiny seeds (200 x $250\mu\textrm{m}$) germinate in response to chemical stimuli produced by host and some non-host plants. Successful contact with their host leads to development of haustoria for obtaining water and food. The shoots above the ground expose flowers and disseminate seeds. Several samples of Orobanche ramosa, O. crenata, O. cernua, and O. egyptiaca were collected from different localities in Jordan. These samples showed one of the following disease symptoms: dry rot at the base of the stem; general deterioration and expanded lesion from base upward; soft tissue maceration of stem; and black rot of flower parts with incomplete maturation of the ovary and seeds. Isolation from diseased stems and seeds was made on three different mycological media. Several fungi were isolated, mainly, Fusarium spp., Alternaria alternata, Rhizoctonia sp., Dendrophora sp., Chaetomium sp., and an ascomycetus fungus with a perithecium. Pathogenicity tests showed that Fusarium spp. and Alternaria alternata attacked healthy living tissue of Orobanche spikes. These fungi caused lesions of black soft rot and complete deterioration within 5-7 days. They also attacked Orobanche seeds, arresting their germination and causing maceration of non-germinated and germinated seeds after 5-7 days of incubation. Meanwhile, Dendrophora sp. and Chaetomium sp. caused limited lesion at first, but were able to colonize the tissue as it aged and senesced. This study showed the presence of a potential endogenous pathogenic fungi in Jordan, which can be investigated as a biological control for Orobanche.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.123-128
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• 2001

In order to develop the biotechnological methods for the mass production of anticancer compounds from tissue culture of Panax ginseng C.A. Mayer, these studies were carried out for the selection of ginseng cell lines containing higher concentration of polyacetylene compounds and optimal condition for their biosynthesis. Panaxynol, one of ginseng polyacetylene, was not detected in any callus induced from ginseng superior cell lines cultured on MS medium supplemented with $\beta$-chlorophenoxy acetic acid (CPA). Panaxydol, another one of polyacetylene and anticancer compounds, were detected in calli of 5 cell lines by thin layer chromatogram and gas chromatogram. Among the 18 ginseng superior lines, the cell line 30201 has higher content of panaxydol. Especially, panaxydol was not detected in the callus induced from cell line 10301 which cultured on the medium containing CPA only, however, it was detected on the same callus cultured on mixed medium containing CAP 2 mg/L and BA 0.05 mg/L. SH medium was better than MS medium for ginseng callus growth and biosynthesis of polyacetylene, and also found that it was not effected by NAA and sucrose concentration in the culture medium.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.53-59
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• 2010

A specific deleted version of ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) lacking the signal receiver domain (1.152 amino acids)-coding sequence, referred to as $ARR1{\Delta}DDK$, was amplified using Arabidopsis thaliana cDNA prepared from adult leaves and transferred into the genome of Nicotiana tabacum cv. Samsun under the transcriptional control of a ${\beta}$-estradiol-inducible expression system. The ectopic expression of $ARR1{\Delta}DDK$ affected the morphology of transgenic seedlings and their segments in vitro. In the presence of an inducer, ${\beta}$-estradiol, ectopic expression of $ARR1{\Delta}DDK$ induced only the formation of soft, pseudo-bulbous tissue in the root tip region of intact seedlings, which appeared similar to callus generated on a hypocotyl segment in the presence of 2,4-D and 6-benzyladenine (BA), both at $1\;{\mu}M$. Those callus tissues on the root tip region could not generate shoots unless $1\;{\mu}M$ BA was supplied. In segment culture, ectopic expression of $ARR1{\Delta}DDK$ induced calluslike tissue around the cut-end of cotyledon and hypocotyl segments with occasional shoot formation, suggesting that the expression of $ARR1{\Delta}DDK$ could substitute for the effects of cytokinin on these segments. Additionally, treatment with only ${\beta}$-estradiol induced NtWUS, a WUS ortholog in tobacco, which was detected during the process of callus tissue formation in the root tip region and also in cotyledon or hypocotyl segments. These findings suggest that the NtWUS might be associated in the transdifferentiation process caused by the functional regulation of $ARR1{\Delta}DDK$ in transgenic tobacco seedlings.