• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.31-35
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• 1999

Morphological characteristic during formation of tobacco crown gall tumor and genetic tumor, and their differential response to growth regulator were investigated in in vitro culture. Crown gall tumor was induced from tumor tissue transformed by infecting Agrobacterium tumefaciens C58. Genetic tumor was induced from tumor tissue which was induced spontaneously from reciprocal interspecific hybrids between Nicotiana glauca (2n=24) and Nicotiana langsdorffii (2n=18). Morphological characteristic of crown gall tumor, genetic tumor, and teratoma shoot was very similar, and they were actively proliferated on hormone-free medium. Typical tumor callus and teratoma shoot formed from crown gall tumor on the hormone-free medium. On the contrary, tumor callus derived from genetic tumor formed as a crown gall tumor callus on the medium supplemented with 0.5 mg/L of 2,4-D, and lots of teratoma shoots without any root formed on the hormone-free medium. Root development from the teratoma shoots was hardly obtained on the medium with IAA, GA and active carbon. However, teratoma shoots with roots, as normal shoots, were initiated occasionally on the hormone-free medium. These shoots also formed new genetic tumor on the stem, which leaves formed lots of teratoma shoot on the hormone-free medium in in vitro culture.

• The Plant Pathology Journal
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• v.30 no.1
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• pp.75-81
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• 2014

In total, 170 carrot lines developed in Korea were screened for resistance to Meloidogyne incognita race 1 to select parental genetic resources useful for the development of nematode-resistant carrot cultivars. Using the gall index (GI), gall formation was examined on carrot roots inoculated with approximately 1,000 second-stage juveniles of the nematode 7 weeks after inoculation. Sixty-one carrot lines were resistant (GI ${\leq}1.0$), while the other 109 were susceptible (GI > 1.0) with coefficient of variance (CV) of GI for total carrot lines 0.68, indicating low-variation of GI within the lines examined. The histopathological responses of two carrot plants from resistant and susceptible lines were examined after nematode infection. In susceptible carrots, giant cells formed with no discernible necrosis around the infecting nematodes. In the resistant carrot line, however, no giant cells formed, although modified cells were observed with extensive formation of necrotic layers through their middle lamella and around the infecting nematodes. This suggested that these structural modifications were related to hypersensitive responses governed by the expression of true resistance genes. Therefore, the Korean carrot lines resistant to the nematode infection are potential genetic resources for the development of quality carrot cultivars resistant to M. incognita race 1.

• Research in Plant Disease
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• v.14 no.3
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• pp.159-164
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• 2008

Crown gall of rose (Rosa hybrida) was observed in greenhouse during 2003-2007. The average disease incidence was up to 38.1 % and was more severe in hydroponic culture as compared to soil culture. The typical gall symptom occurred mainly on the root, crown, or both, and resulted on poor rooting, growth retardation and yield loss. The reduction rate of rooting influenced by crown gall was 57.5% as compared to healthy plants on nursery stock. The location of gall formation in the plant influenced growth vigor resulting in symptoms such as poor shooting. Healthy plants produced 19.1 flowers/$m^2$, while diseased plants produced 9.5 flowers/$m^2$ during the same cultivation period. Moreover, the number of days to flowering was longer for the diseased plants than for healthy plants - 51.2 days and 39.8 days for first harvest, and 60.6 days and 52.1 days for the second harvest, respectively. Conclusively, infection on the basal stem caused serious loss of the number of shoot formation; yield loss of cut flower was 38.7% due to crown gall infection and delay of harvesting time about 8-10 days.

• Research in Plant Disease
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• v.12 no.3
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• pp.197-204
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• 2006

Crown gall on lower stem and root of chrysanthemum(Dendranthema grandiflorum Kitamura) was observed at Hwasung and Gumi in 2001 and 2004, respectively. Tumors were semi-round with rough surface texture of dark brown color. Nine isolates inducing gall formation on lower stem of chrysanthemum among twenty isolates from the tumor tissues were characterized. Their colonies were convex, glistening, circular with an entire edge and whitish or tannish cream in color on potato dextrose agar supplemented with 0.5% $CaCO_3$. The virulent isolates were rod-shaped with peritrichous flagellae, gram negative, aerobic and growing on D1M agar. Among nine virulent isolates, one isolate was identified as Agrobacterium rubi and eight isolates were A. tumefaciens based on biochemical and physiological characteristics, fatty acid profiles and substrate utilization patterns. A. tumefaciens had strong pathogenicity and broad host range compared with A. rubi. This is the first report on crown gall of chrysanthemum in Korea. To our knowledge, crown gall of chrysanthemum caused by A. rubi is first report in this study worldwide.

• The Plant Pathology Journal
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• v.33 no.4
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• pp.410-423
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• 2017

Pathological interrelations of two soil-borne diseases in cucurbits (watermelon, oriental melon, shintosa and cucumber) caused by Fusarium isolates (FI) and the root-knot nematode (RKN), Meloidogyne incognita were characterized by the fusarium disease severity index (DI), RKN gall index (GI) and eggmass index (EI) in inoculation tests using FI and RKN. Virulence of FI as determined by DI at 4 weeks after inoculation was mostly in the higher order of Fusarium proliferatum F6, F5 and Fusarium oxysporum f. sp. melonis or Fusarium oxysporum f. sp. niveum with no significant differential interactions among the cucurbits and RKN co-infection. Significant increases of DI due to RKN coinfection were noticed in watermelon and oriental melon infected with F. proliferatum isolates, suggesting the DI increase due to RKN coinfection may depend upon the virulence of FI relative to aggressiveness of RKN on the cucurbits. For the coinfection of FI and RKN, GI and EI were mostly reduced logarithmically with the increase of DI, largely more in EI than GI, in all cucurbits except for shintosa. Microscopic examination of the root tissues showed histopathological features characteristic to infection types; formation of fungal hyphae and/or spores and plant defense structures (tyloses and mucilage) in variable degrees and formation of giant cells at variable developmental stages and with variable cytoplasmic depletion or degeneration which were visualized in relations with the values of DI, GI and EI. These findings will be helpful to develop control strategies of the soil-borne disease complex based on their pathological characteristics.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.149-153
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• 2010

Biological and structural mechanisms of the nematode disease development in chili pepper caused by the root-knot nematode, Meloidogyne incognita, were investigated. Out of 39 chili pepper cultivars/lines tested, six were found resistant, while 33 were susceptible to M. incognita, of which a susceptible cultivar Chilseongcho and three resistant cultivar/lines CM334, 02G132 and 03G53 with different resistance degrees were selected for microscopic studies on the disease development. Gall formation was greatly reduced in the resistant cultivars/lines. Nematode penetration occurred both in the susceptible and resistant chili pepper roots; however, the penetration rates were significantly lowered in the three resistant peppers compared to the susceptible pepper cv. Chilseongcho. In the susceptible pepper, giant cells were extensively formed with no discernible necrosis around the nematode feeding sites. In the highly resistant pepper cultivar CM334, no giant cell was formed, but extensive necrosis formation was observed around the penetrating nematodes. In the other two resistant pepper lines (02G132 and 03G53), both giant cells and prominent necroses were formed, and the necrotic responses appeared to inhibit the further development of giant cells or accelerate their early degeneration. Although the nematode penetration was retarded significantly in the resistant cultivar/lines, all of the above results suggest that the disease resistance of pepper may be related to post-infectional defense mechanisms (nematode growth and development) more than pre-infectional ones (penetration and establishment). Variations in structural modifications in the resistant cultivar/lines may reflect their genetic differences related to the nematode resistance.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.960-966
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• 2022

Lactic acid bacteria (LAB) exert antagonistic activity against root-knot nematodes, mainly by producing organic acids via carbohydrate fermentation. However, they have not yet been used for root-knot nematode (Meloidogyne incognita) control owing to a lack of economic feasibility and effectiveness. In this study, we aimed to isolate organic acid-producing LAB from kimchi (Korean traditional fermented cabbage) and evaluated their nematicidal activity. Among the 234 strains isolated, those showing the highest nematicidal activity were selected and identified as Lactiplantibacillus plantarum WiKim0090. Nematicidal activity and egg hatch inhibitory activity of WiKim0090 culture filtrate were dose dependent. Nematode mortality 3 days after treatment with 2.5% of the culture filtrate was 100%, with a 50% lethal concentration of 1.41%. In pot tests, the inhibitory activity of an L. plantarum WiKim0090-copper sulfate mixture on gall formation increased. Compared to abamectin application, which is a commercial nematicide, a higher control value was observed using the WiKim0090-copper sulfate mixture, indicating that this combination can be effective in controlling the root-knot nematode.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.968-975
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• 2018

In the course of screening for microbes with nematicidal activity, we found that Enterobacter asburiae HK169 displayed promising nematicidal activity against the root-knot nematode Meloidogyne incognita, along with plant growth-promoting properties. Soil drenching of a culture of HK169 reduced gall formation by 66% while also increasing root and shoot weights by 251% and 160%, respectively, compared with an untreated control. The cell-free culture filtrate of the HK169 culture killed all juveniles of M. incognita within 48 h. In addition, the nematicidal activity of the culture filtrate was dramatically reduced by a protease inhibitor, suggesting that proteolytic enzymes contribute to the nematicidal activity of HK169. In order to obtain genomic information about the HK169 isolate related to its nematicidal and plant growth-promoting activities, we sequenced and analyzed the whole genome of the HK169 isolate, and the resulting information provided evidence that the HK169 isolate has nematicidal and plant growth-promoting activities. Taken together, these observations enable the future application of E. asburiae HK169 as a biocontrol agent for nematode control and promote our understanding of the beneficial interactions between E. asburiae HK169 and plants.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.662-673
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• 2019

Root-knot nematodes (RKNs) are among the most destructive plant-parasites worldwide, and RKN control has been attempted mainly using chemical nematicides. However, these chemical nematicides have negative effects on humans and the environment, thus necessitating the search for eco-friendly alternative RKN control methods. Here, we screened nematicidal lactic acid bacteria (LAB) isolated from kimchi and evaluated their efficacy as biocontrol agents against RKNs. Of 237 bacterial strains, Lactobacillus brevis WiKim0069 showed the strongest nematicidal activity against the second-stage juveniles (J2) of Meloidogyne incognita, M. arenaria, and M. hapla and inhibited the egg hatch of M. incognita. The culture filtrate of WiKim0069 had a pH of 4.2 and contained acetic acid (11,190 ㎍/ml), lactic acid (7,790 ㎍/ml), malic acid (470 ㎍/ml), and succinic acid (660 ㎍/ml). An artificial mixture of the four organic acids produced by WiKim0069 also induced 98% M. incognita J2 mortality at a concentration of 1.25%, indicating that its nematicidal activity was derived mainly from the four organic acids. Application of WiKim0069 culture filtrate suppressed the formation of galls and egg masses on tomato roots by M. incognita in a dose-dependent manner in a pot experiment. The fermentation broth of WiKim0069 also reduced gall formation on melon under field conditions, with a higher efficacy (62.8%) than that of fosthiazate (32.8%). This study is the first report to identify the effectiveness of kimchi LAB against RKNs and to demonstrate that the organic acids produced by LAB can be used for the RKN management.