• Journal of Crop Science and Biotechnology
• /
• v.11 no.2
• /
• pp.91-100
• /
• 2008

Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

• Research in Plant Disease
• /
• v.23 no.4
• /
• pp.348-354
• /
• 2017

To investigate occurrence of root-knot nematode (RKN) in plastic house of central area of Korea, 132 soil samples were collected in cucumber, water melon, tomato, red pepper and strawberry fields from 2013 to 2015. Among 132 soil samples, 65 soil samples (49%) were infested with RKN and mean density of RKN was 178 second-stage juveniles per $100cm^3$ soil (min. 1 ~ max. 3,947). The frequency of RKN by regional was the highest in Chuncheon with 80%, followed by Cheonan (68%), Nonsan (36%), Buyeo (33%) and Yesan (30%). The frequency of RKN by crops was the highest in tomato with 83%, followed by cucumber (61%), strawberry (41%), red pepper (30%), watermelon (26%). To identify the species of RKN, fifteen populations were selected for representative populations. As a phylogenetic analysis of 15 populations, southern root-knot nematode (Meloidogyne incognita), peanut root-knot nematode (M. arenaria) and northern root-knot nematode (M. hapla) were identified with 47%, 20% and 33% ratio, respectively. In crops, M. incognita, M. arenaria and M. hapla were detected in tomato, M. incognita and M. arenaria were detected in cucumber and watermelon, and M. hapla was detected in strawberry and lettuce. Thus, there should be a continuous management to major species of each crops to prevent dispersal of RKN damages.

• Research in Plant Disease
• /
• v.7 no.1
• /
• pp.49-55
• /
• 2001

Species and races of root-knot nematodes in greenhouses in southern Korea were investigated and a simple identification scheme was provided. Among 23 populations of root-knot nematodes, Meloidogyne arenaria race 2 was 59%, M. incognita race 1 was 23%, and an unknown race of M. incognita was 18%. Total length of M. arenaria juveniles was 411㎛(306-503㎛) and that of M. incognita was 384㎛(312-488㎛); however, the ranges of two species were overlapped and could not be used to distinguish the two species. Excretory pore in female head was a consistent character to differentiate M. arenaria and M. incognita.

• The Plant Pathology Journal
• /
• v.19 no.3
• /
• pp.177-180
• /
• 2003

Pasteuria penetrans KW1 (PP), parasitic bacterium of nematode, was isolated from oriental melon greenhouse soil in Korea and evaluated for the suppression effect on the reproduction of southern root-knot nematode, Meloidogyne incognita (MI), in lettuce (Lactuca sativa L. var. Chungchima) and oriental melon (Cucumis melo L. var. Eunchun). Pot experiments were conducted by planting the lettuce seedlings in medium inoculated with 5,000 MI juveniles/pot (J), J +100,000 PP endospores/l g medium, and J +200,000 PP endospores/1 g medium. After 11 weeks of plantation, number of root galls in J +200,000 PP endospores/1 g medium was decreased to 92/root (38.9%, control effect), compared to the J of 150/root. In the second plantation of lettuce in the same pots, the numbers of root gall were significantly decreased in PP treated pots with 75 (77.2%, control effect) and 150/root (54.4%, control effect) in J +200,000 and J +100,000 PP endospores/1 g medium, respectively, compared to the J of 330/root when harvested at 10 weeks after planting. In oriental melon, root gall percentages were 32.1 (60.2%, control effect) and 52.9％ (34.5%, control effect) in J +200,000 and J + 1(10,000 endospores/l g medium which were significantly lower than that of 80.7％ in J.

• Korean journal of applied entomology
• /
• v.26 no.4 s.73
• /
• pp.195-201
• /
• 1987

Insects and nematodes associated with crops growing in plastic film houses were surveyed throughout the southern part of Korea at Jinju, Jiphyeon, Geumsan, Hapcheon, Changyeong, Namji, Milyang, Kimhae, Busan, Sooncheon and Gwangyang from December of 1984 to December of 1985. The phytonematodes representing six families, nine genera and nine species, and the insects representing four orders, seventeen families, twenty-three genera and twenty-four species were identified. The mite also caused problem on the leaves of strawberry and watermelon. Of these Meloidogyne incognita and Aphis gossyphii were most important ones. Aphelenchoides fragariae and M. hapla, however dominant nematodes on strawberry. M. incognita was always detected from pepper plants which were heavily infected with Phytophthora capsici. Tomato roots were readily infected with root-knot nematodes in non-sterilized upland surface soil. However, tomato were growing-vigorouly free from nematode damage in the upland surface soil treated by nematicide or in the paddy soil. A few galls were developed even in the upland subsoil at 60cm below surface. Soil salinity affected profoundly the host-root-knot nematode interaction: the numerous galls were developed on the tomato roots at EC $1mS/cm^2$ followed by 4mS and 2mS, but few galls were at 6mS.

• Korean journal of applied entomology
• /
• v.52 no.4
• /
• pp.327-333
• /
• 2013

Plant parasitic nematodes were isolated from 274 soil samples collected from soybean fields in Korea. Nematode importance rating in soybean is proposed based on this study and by reviewing other reports. Soybean cyst nematode, Heterodera glycines is the most important nematode species and rated as $1^{st}$ because it detected from 38%(range 25-51%) of soil samples with high density except Jeju province. Root-knot nematode, Meloidogyne spp. is rated $2^{nd}$ and is more widely distributed in southern provinces, Gyeongnam, Jeonnam, and Jeju province (detection rate ranged 16-44%). Pratylenchus is rated $3^{rd}$ and is more frequently detected from northern provinces such as Gyeonggi and Gangwon (detection rate ranged 10-13%). Helicotylenchus is rated $4^{th}$ and is particularly important in Jeju province (detection rate is 62% and numbers averaged 571 nematodes/$300cm^3$ soil), which is the main production area for sprouting soybean in Korea. Tylenchorhynchus, Paratylenchus, Criconema, Criconemoide, Mesocriconema, Ogma, Xiphinema, Paratrichodorus, and Trichodorus occur in low frequency and density, thus they are rated to $5^{th}$. We propose to delete following nematode genus from the list of soybean parasitic nematode in Korea because their parasitism on soybean is unconfirmed or negligible; Tylenchus, Aphelenchus, Aphelenchoides, Ditylenchus, Hirschmanniella immamuri, Basiria graminophila, Psilenchus, and Pseudhalenchus. Therefore, we revised 30 species in 16 genera as soybean parasitic nematodes in Korea. Importance rating is $1^{st}$ Heterodera glycines, $2^{nd}$ Meloidogyne spp., $3^{rd}$ Pratylenchus spp. and $4^{th}$ Helicotylenchus spp. especially in Jeju province.