• Journal of Life Science
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• v.19 no.5
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• pp.639-643
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• 2009

High temperature stress might affect the yield and quality of Chinese cabbage. In order to develop cultivars resistant to high temperature stress, we developed polymorphic DNA markers for trehalose synthesis genes related to abiotic stress resistance. A total of 28 Brassica rapa ESTs homologous to trehalose synthesis genes of Arabidopsis were found from the NCBI database. The polymorphic DNA sequences were searched between Chinese cabbages - Chiifu, which is relatively susceptible to high temperature stress, and Kenshin, which is tolerant to high temperature stress. Among the 28 ESTs, we found 10 ESTs that have either insertion/deletion and/or single nucleotide polymorphism between the two cultivars. Those polymorphic sites were then targeted for the development of 10 PCR based markers. These molecular markers related to trehalose genes could be used not only to test their relationship with abiotic stress resistance in Chinese cabbage, but also the development of abiotic stress resistant cultivars using MAS.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.29-29
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• 2015

Clubroot disease is caused by the soil-born obligate plant pathogen Plasmodiophora brassicae. This pathogen can infect all cruciferous vegetables and oil crops, including Brassica rapa, B. oleracea, B. napus, and other Brassica species. Clubroot disease is now considered to be a major problem in Chinese cabbage production in China, Korea, and Japan. We collected several hundreds of P. brassicae infected galls from Korea, and isolated the single spore from the collection. For establishment of novel isolation, and mass-propagation methods for singe spore isolates of P. brassicae pathogen, we developed new filtration method using both cellulose nitrate filter and syringe filter. Accurate detection of P. brassicae pathogen in the field was done by using real-time PCR in the potential infested soil. When we tested the different pathogenicity on commercial Chinese cabbage varieties, P. brassicae from collected galls showed various morphological patterns about clubroot symptom on roots. To date, 8 CR loci have been identified in the B. rapa genome using the quantitative trait loci (QTL) mapping approach, with different resistant sources and isolates. We are trying to develop the molecular marker systems for detect all 8 CR resistant genes. Especially for the study on the interaction between pathogens and CR loci which are not well understood until now, genome wide association studies are doing using the sequenced inbred lines of Chinese cabbage to detect the novel CR genes.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.49-54
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• 2013

C3HC4-type RING zinc finger proteins essential in the regulation of plant processes, including responses to abiotic stresses. We previously isolated and examined the C3HC4-type RING zinc finger protein (BrRZFP1) from Brassica rapa under abiotic stresses. To elucidate the role of the BrRZFP1 transcription factor in gene regulation, we transformed tobacco plants with the BrRZFP1 gene. Plants were regenerated from 82 independently transformed callus lines of tobacco and analysed for transgene expression. Transgene integration and expression was confirmed by Southern and RT-PCR analyses, respectively. T2 plants displayed more tolerance to the bacterial pathogens Pectobacterium carotovorum and Ralstonia solanacearum, and the tolerance levels were correlated with BrRZFP1 expression levels. These results suggest that the transcription factor BrRZFP1 is an important determinant of stress response in plants and its overexpression in plants could increase biotic stress resistance.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.625-629
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• 1998

Turnip mosaic potyviruses (TuMV) were isolated from Rorippa indica and Armoracia lapathifolia showing mosaic symptoms in field. Identification of the TuMVs were carried out by host reactions of indicator plants, electron micrograph, serological properties and reverse transcription-poly-merase chain reaction (RT-PCR). Both viruses systemically infected Chenopodium quinoa, Nicotiana clevelandii, Brassica rapa, B. campestris subsp. pekinensis, B. juncea and Raphanus sativus, and developed local infection on inoculated leaves of C. quinoa, C. amaranticola, C. album, N. tabacum cv. Xanthi nc and Gomphrena grobosa. However, the viruses did not infect on N. glutinosa, Cucumis sativus and Vigna unguiculata. The filamentous particles, about 720 nm in length, and inclusion bodies were observed from the infected leaf tissues by dipping on electron microscopy. Crude sap of leaf infected with the viruses was reacted positively with an antiserum of TuMV in agar gel double diffusion. For detection of the viruses, RT-PCR was carried out with TuMV--specfic oligonucleotide primer. The RT-PCR products, a 1,092 bp DNA fragment, were obtained from naturally infected leaves of R. indica and A. lapathifolia. In inoculation test to seven cruciferous weeds with TuMV, infection occurred in Arabis glabra, Barbarea orthoceras, Capsella bursa-pastoris, Draba nomorosa var. hebecarpa, Rorippa cantoniensis and Thlaspi arvense.

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.33-48
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• 2012

As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant $Arabidopsis$ was available. As a result, the whole $B.$ $rapa$ genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in $Brassica$ crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of $Brassica$ genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within $Brassica$ as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in $Brassica$ crop.

• Journal of fish pathology
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• v.18 no.1
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• pp.39-48
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• 2005

Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

• Research in Plant Disease
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• v.17 no.2
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• pp.211-215
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• 2011

Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.209-214
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• 2003

Expression of 43 flowering-related genes was examined in two inbred lines of Chinese cabbage, Chiifu and Kenshin, under different photoperiod, vernalization and flower development stages. The floral genes cloned by RT-PCR with degenerated primers showed high homology with Arabidopsis counterparts. Genes in two inbred lines, TOC, CRY1, CO, RGAL and GAl, were highly expressed under all tested conditions. However, expression of three genes was regulated by particular experimental conditions. The expression of LHY gene was predominant in Chiifu under the short-day conditions, whereas the expression of RGAL gene was influenced by vernalization in both inbred lines. Besides, the expression of NAP gene was induced by vernalization only in Chiifu. Most of the flower identity-related genes were expressed during flower development. The transcript level for several genes was not detected in this experiment.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.33-33
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• 2015

Clubroot disease is the major threat to the production of Chinese cabbage (Brassica rapa L.) in Japan. Although the breeding of the clubtoot resistant (CR) cultivars is one of the most efficient ways to control this disease, the CR cultivars do not always have effects due to the breakdown of resistance. Therefore, it is necessary to develop the breeding strategy to accumulate multiple CR genes in a single cultivar effectively. We have identified two incomplete dominant CR loci, Crr1 and Crr2, which are originated from the European CR turnip Siloga. To investigate the effectiveness of marker-assisted selection (MAS) for CR breeding, the inbred line with Crr1 and Crr2 was crossed with parental lines of the existing CR $F_1$ cultivar of Chinese cabbage, followed by 5 times of MAS and backcrossing. The $F_1$ derived from a cross between the resulting parental lines improved the clubroot resistance as expected and had the same morphological characters as the original $F_1$ cultivar. We have shown that the Crr1 locus comprised two loci: Crr1a, which by itself conferred resistance to the mild isolate; and Crr1b, which had a minor effect, but was not required for Crr1a-mediated resistance. Further genetic analysis suggested that Crr1b was necessary to acquire resistance to the more virulent isolate in combination with Crr2. Molecular characterization of Crr1a encoding TIR-NB-LRR class of R protein revealed that there were at least 4 alleles in Japanese CR cultivars of Chinese cabbage. PCR analysis with Crr1a-specific markers demonstrated that the functional alleles were predicted to be present in European CR turnips, Debra and 77b besides Siloga, whereas rarely in Japanese CR cultivars, indicating that Crr1a is an useful source to improve the resistance of Chinese cabbage cultivars.

• Plant Biotechnology Reports
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• v.5 no.3
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• pp.217-224
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• 2011

Non-heading Chinese cabbage (Brassica rapa L. ssp. chinensis) is a popular vegetable in Asian countries. The diamondback moth (DBM), Plutella xylostella (L.), an insect with worldwide distribution, is a main pest of Brassicaceae crops and causes enormous crop losses. Transfer of the anti-insect gene into the plant genome by transgenic technology and subsequent breeding of insect-resistant varieties will be an effective approach to reducing the damage caused by this pest. We have produced transgenic non-heading Chinese cabbage plants expressing the potato proteinase inhibitor II gene (pinII) and tested the pest resistance of these transgenic plants. Non-heading Chinese cabbages grown for 45 days on which buds had formed were used as experimental materials for Agrobacterium-mediated vacuum infiltration transformation. Forty-one resistant plants were selected from 1166 g of seed harvested from the infiltrated plants based on the resistance of the young seedlings to the herbicide Basta. The transgenic traits were further confirmed by the Chlorophenol red test, PCR, and genomic Southern blotting. The results showed that the bar and pinII genes were co-integrated into the resistant plant genome. A bioassay of insect resistance in the second generation of individual lines of the transgenic plants showed that DBM larvae fed on transgenic leaves were severely stunted and had a higher mortality than those fed on the wild-type leaves.