• The Plant Pathology Journal
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• v.25 no.4
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• pp.369-375
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• 2009

The potential antiviral effects of the culture filtrates (CF) from Serratia marcescens strain Gsm01 against yellow strain of Cucumber mosaic virus (CMV-Y) were investigated. The culture filtrate of S. marcescens strain Gsm01 applied on Chenopodium amaranticolor showed high inhibitory activity, likewise no necrosis appeared when applied on the tobacco plants 2 days before CMV-Y inoculation. When plants were challenge inoculated with CMV-Y for eighteen days, the disease incidence in plants with culture filtrate of S. marcescens Gsm01 did not exceed 59%, whereas 100% of control plants were severely infected. The results of double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA), reverse transcriptase polymerase chain reaction (RT-PCR), dot blotting, and western blotting showed that culture filtrate treatment highly affected the accumulation of CMV-Y or its CP protein gene in the treated plant leaves. It was also observed that the culture filtrate had no RNase activity on genomic RNAs of CMV-Y, suggesting that culture filtrate may not contain ribosome inactivating proteins (RIPs) or proteins with RNase activity. These data shows that culture filtrate of S. marcescens strain Gsm01 seems to be a promising source of antiviral substance for the practical use.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.453-461
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• 2016

Background: This study aims to describe the characterization of Paenibacillus polymyxa GBR-1 (GBR-1) with respect to its positive and negative effects on plants. Methods: The morphological characteristics of GBR-1 were identified with microscopy, and subjected to Biolog analysis for identification. Bacterial population and media optimization were determined by a growth curve. The potential for GBR-1 as a growth promoting agent, to have antagonistic activity, and to have hydrolytic activity at different temperatures was assessed. The coinoculation of GBR-1 with other microorganisms and its pathogenicity on various stored plants, including ginseng, were assessed. Results: Colony morphology, endospore-bearing cells, and cell division of GBR-1 were identified by microscopy; identification was performed by utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME). GBR-1 showed the strongest antagonistic activity against fungal and bacterial pathogens. GBR-1 cell numbers were relatively higher when the cells were cultured in brain heart infusion (BHI) medium when compared with other media. Furthermore, the starch-hydrolytic activity was influenced by GBR-1 at higher temperature compared to low temperatures. GBR-1 was pathogenic to some of the storage plants. Coinoculation of GBR-1 with other pathogens causes differences in rotting on ginseng roots. A significant growth promotion was observed in tobacco seedlings treated with GBR-1 suspensions under in vitro conditions, suggesting that its volatile organic compounds (VOCs) might play a role in growth promotion. Conclusion: The results of this study indicate that GBR-1 has both positive and negative effects on ginseng root and other stored plants as a potential biocontrol agent and eliciting in vitro growth promotion.

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.3
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• pp.281-290
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• 1999

The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with 35S promoter of cauliflower mosaic virus (CaMV) and introduced into tobacco plants (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. As expected, ipt gene was constitutively expressed in all tissues of transgenic plants. Several primary transgenic plants were obtained that expressed different level of transcripts for ipt gene. Three of transgenic plants with different expression level of ipt gene were selected and selfed to obtain homozygous line for further analysis. A number of interesting phenotypic changes such as viviparous leaves, delayed senescence, larger axillary shoots, an abundance of tiny shoots at the apex and a release of lateral buds, were observed in transgenic plants. Chlorophyll content was 1.5- t.o 4-fold higher in transgenic plants as compared with non-transformed plants. These results indicate that the cytokinin synthesized in transgenic plants could improve forage crop yield by delay of leaf senescence and increase of leaf number.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.61-71
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• 2000

By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.596-602
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• 2019

In September 2017, the rhizospheric soil of Tetragonia tetragonoides (Pall.) Kuntze was further sampled. One hundred and thirty eight species of microorganisms were isolated from the soil. Indole-3-acetic acid (IAA) production, siderophore production, and phosphate degradation were examined in order to confirm bacterial growth from isolated microorganisms. As a result, most strains were able to produce auxins or siderophores and to solubilize phosphate. In addition, 138 isolated strains were treated with tobacco extract and conferred pathogen resistance to host plants upon treatment. As a result, 35 strains that were able to reduce pathophysiology by more the 60% were selected. Among them, 6 strains with high induced systemic resistance (ISR) activity were found. All of these strains belong to the genus Bacillus according to the 16S rDNA sequence analysis. Bacillus aryabhattai KUDC6619 showed outstanding effects with reduced infection in tobacco and pepper plants. Probably, these Bacillus species play a beneficial role by association with T. tetragonoides for its survival in the harsh conditions found on the island of Dokdo.

• Journal of Life Science
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• v.12 no.1
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• pp.96-105
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• 2002

To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.113.2-114
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• 2003

Five pepper-infecting potyviruses, Pepper mottle virus (PepMoV), Chilli veinal mottle virus (CVMV), Pepper veinal mottle virus (PVMV), Pepper severe mosaic virus (PSMV) and Tobacco each virus (TEV), are known filamentous virus and can be infected pepper crops systemically. To understand pathology and genome information of the five viruses on pepper plants, host reactions and sequences were compared to the 5 viruses. Five potyviruses were inoculated onto some typical cultivars of hot peppers and compared their symptoms, and virus accumulations. A set of degenerate primers for potyviruses were applied to 5 viruses and RT-PCR was performed. RT-PCR products containing partial nuclear inclusion b and coat protein (CP) genes were cloned. Then, oligo dT primer and species-specific primer were redesigned to amplify the C-terminal part of CP and 3' noncoding regions of each viruses. Sequences of the viruses were analyzed and compared to serological relationships among the viruses. The data can be useful for screening of potyviruses in pepper plants and pathogen-derived transgenic pepper plant development.

• Mycobiology
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• v.45 no.4
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• pp.263-269
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• 2017

The genus Peronospora, an obligate biotrophic group belonging to Oomycota, causes serious damage to a variety of wild and ornamental plants, as well as cultivated crops, such as beet, rose, spinach, and tobacco. To investigate the diversity of Peronospora species parasitic to Stellaria and Pseudostellaria (Caryophyllaceae) plants in Korea, we performed a morphological analysis on dried herbarium specimens and molecular phylogenetic inferences based on internal transcribed spacer rDNA and cox2 mitochondrial DNA sequences. As a result, it was confirmed that there are four species of Peronospora parasitic to specific species of Stellaria and Pseudostellaria, all of which were hitherto unrecorded in Korea: P. alsinearum (ex Stellaria media), P. stellariae-aquaticae (ex Stellaria aquatica), P. stellariae-uliginosae (ex Stellaria alsine), and P. pseudostellariae (ex Pseudostellaria palibiniana). In addition, Peronospora specimens parasitic to Pseudostellaria davidii differed morphologically from P. pseudostellariae owing to the large and ellipsoidal conidia; this morphological discrepancy was also validated by the high genetic divergence between the two species. Peronospora casparyi sp. nov. is described and illustrated here.

• Journal of Life Science
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• v.18 no.8
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• pp.1023-1030
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• 2008

NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

• Research in Plant Disease
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• v.9 no.2
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• pp.89-93
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• 2003

An isolate of Tobacco mosaic virus (TMV) was isolated from potato cultivar ‘Chubak’ showing vein clearing and mild mosaic at Namhae in Korea. This isolate, TMV-St, was differentiated from other tobamoviruses based on biological properties, serological relationships and nucleotide sequence analyses of coat protein genes. TMV-St caused typical symptoms on four indicator plants as compared to the tobamovirus of TMV-U1, Pepper mild mottle virus (PMMoV), and Tomato mosaic virus (ToMV), which caused economic losses in Solanaceous vegetables, tomato, pepper, and eggplant. Remarkably, the TMV-St induced distinctly different symptom of systemic chlorotic spots on Chenophodium murale. On C. murale, Gomphorena globosa, and Nic-otiana rustica, the four viruses were classed by the virulence of systemic or local infections. In serological test TMV-St antiserum showed a precipitation line with each other tabamovirus. The CP gene of TMV-St contain 477 nucleotides, and the nucleotides sequence was the most similar to that of TMV-U1.