• Research in Plant Disease
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• v.16 no.3
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• pp.279-284
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• 2010

Clubroot caused by Plasmodiophora brassicae is a widespread disease that causes serious problems in many brassica growing areas. To establish more simple and reliable clubroot screening method of Chinese cabbage to P. brassicae using soil-drenching inoculation, the development of clubroot on Chinese cabbage according to several conditions such as soil type, inoculum concentration of P. brassicae GN-1 (race 9), plant growth stage and incubation period was studied. In a commercial horticulture nursery media soil (CNS), disease severity of the seedling according to inoculum concentration increased in a dose-dependent manner, but did not in mixture of CNS and upland soil (1:1, v/v). To facilitate and acquire precise result of resistance screening of Chinese cabbage to clubroot, 10-day-old seedlings should be inoculated by drenching the spore suspension of P. brassicae to give inoculum density of $4.0{\times}10^8$ spores/pot. To develop the disease, the inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days, and then cultivated in a greenhouse ($25{\pm}5^{\circ}C$) for five weeks. Under the optimum conditions, 25 clubroot-resistant (CR) and 3 clubroot-susceptible (CS) cultivars were tested for resistance to P. brassicae. All CR cultivars showed very clear resistance response, on the other hand all CS cultivars severly infected with the pathogen. The results suggest that this method is efficient screening method of Chinese cabbage for resistance to clubroot disease.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.350-355
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• 2015

Clubroot disease is one of the most wide-spread and devastating diseases in the cultivation of Chinese cabbage. To develop a protein marker for resistance to clubroot disease in Chinese cabbage, a comparative proteome analysis was performed between a sensitive line, 94SK, and a resistant line, CR Shinki DH. Three proteins of two fold or higher accumulation that are specific to each line were found 3 days after innoculation of the Plasmodiphora brassicae. They are glutamine synthetase, malate dehydrogenase/oxidoreductase and fructose-bisphosphate aldolase in the 94SK and actin, phosphoglycerate kinase, and Cu/Zn superoxide dismutase in the CR Shinki line. From the comparison of the synthesized proteins in the 94SK and the CR Shinki, CR Shinki was found to produce more ATP-binding protein for the ABC transporter while 94SK showed a higher level of pathogenesis-related protein 1 production. All of these proteomic variations may lead to the development of molecular markers to accelerate the breeding process.

• Research in Plant Disease
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• v.20 no.4
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• pp.235-244
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• 2014

Clubroot and Fusarium wilt of cole crops (Brassica oleracea L.) are destructive diseases which for many years has brought a decline in quality and large losses in yields all over the world. The breeding of resistant cultivars is an effective approach to reduce the use of chemical fungicides and minimize crop losses. This study was conducted to evaluate the resistance of 60 cabbage (B. oleracea var. capitata) and 6 broccoli (B. oleracea var. italica) lines provided by The RDA-Genebank Information Center to clubroot and Fusarium wilt. To investigate resistance to clubroot, seedlings of the genetic resources were inoculated with Plasmodiophora brassicae by drenching the roots with a mixed spore suspension (1 : 1) of two isolates. Of the tested genetic resources, four cabbage lines were moderately resistant and 'K166220' represented the highest resistance to P. brassicae. The others were susceptible to clubroot. On the other hand, to select resistant plants to Fusarium wilt, the genetic resources were inoculated with Fusarium oxysporum f. sp. conglutinans by dipping the roots in spore suspension of the fungus. Among them, 17 cabbage and 5 broccoli lines were resistant, 16 cabbage lines were moderately resistant, and the others were susceptible to Fusarium wilt. Especially, three cabbage ('IT227115', 'K161791', 'K173350') and two broccoli ('IT227100', 'IT227099') lines were highly resistant to the fungus. We suggest that the resistant genetic resources can be used as a basic material for resistant B. oleracea breeding system against clubroot and Fusarium wilt.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.41-41
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• 2015

BSA-seq technologies, combined Bulked Segregant Analysis (BSA) and Next-Generation Sequencing (NGS), are making it faster and more efficient to establish the association of agronomic traits with molecular markers or candidate genes, which is the requirement for marker-assisted selection in molecular breeding. Clubroot disease, caused by Plasmodiophora brassicae, is a serious threat to Brassica crops. Even we have breed new clubroot resistant varieties of Chinese cabbage (B. rapa ssp. pekinesis), the underlying genetic mechanism is unclear. In this study, an $F_2$ population of 340 plants were inoculated with P. brassicae from Xinye (Pathotype 2 on the differentials of Williams). Resistance phenotype segregation ratio for the populations fit a 3:1 (R:S) segregation model, consistent with a single dominant gene model. Super-BSA, using re-sequencing the parents, extremely R and S DNA pools with each 50 plants, revealed 3 potential candidate regions on the chromosome A03, with the most significant region falling between 24.30 Mb and 24.75 Mb. A linkage map with 31 markers in this region was constructed with several closely linked markers identified. A Major QTL for clubroot resistance, CRq, which was identified with the peak LOD score at 169.3, explaining 89.9% of the phenotypic variation. And we developed a new co-segregated InDel marker BrQ-2. Joint BSA-seq and traditional QTL analysis delimited CRq to an 250 kb genomic region, where four TIR-NBS-LRR genes (Bra019409, Bra019410, Bra019412 and Bra019413) clustered. The CR gene CRq and closely linked markers will be highly useful for breeding new resistant Chinese cabbage cultivars.

• Horticultural Science & Technology
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• v.29 no.6
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• pp.610-616
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• 2011

Clubroot disease caused by Plasmodiophora brassicae, induces damage to cruciferous vegetables worldwide. For control of the disease, many CR (clubroot resistant) $F_1$ hybrid cultivars of Chinese cabbage have been bred and released in Korea. In this study, we determined the race of 10 field isolates of P. brassicae collected from ten regions in Korea using Williams' differential varieties and investigated the degree of resistance of 25 commercial CR cultivars of Chinese cabbage to the isolates. The clubroot pathogens were assigned into two (HS and YC), two (HN1 and HN2), two (DJ and SS) and four (GS, GN, JS, and PC) isolates for race 2, race 4, race 5, race 9, respectively. All CR cultivars showed similar response, resistant or susceptible, to each isolate and the P. brassicae isolates were divided into two groups. Among them, the DJ, GS, GN, HS, and JS isolates could not infect the CR cultivars. In contrast, the SS, HN1, HN2, PC, and YC isolates caused severe clubroot disease on the CR cultivars like susceptible cultivars. Even though they belong to the same race, the CR cultivars showed a different response to the pathogens. The results suggest that the breakdown of CR in Chinese cabbage has already occurred in cultivation areas of Korea and resistance source introduced in CR cultivars may be very limited. In addition, it is likely that resistance genes of Williams' differential varieties to P. brassicae are different from the gene of CR cultivars of Chinese cabbage used in the study.

• Research in Plant Disease
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• v.17 no.2
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• pp.161-168
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• 2011

To establish simple and reliable screening method for resistant radish to Plasmodiophora brassicae Woron. using soil-drenching inoculation, the development of clubroot on radish seedlings inoculated with P. brassicae GN-1 isolate according to several conditions such as inoculum concentration, plant growth stage and incubation period after inoculation was studied. To select resistant radish against clubroot, 10-day-old seedlings were inoculated with P. brassicae by drenching the roots with the spore suspension of the pathogen to give $1{\times}10^9$ spores/pot. The inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days then cultivated in a greenhouse ($20{\pm}5^{\circ}C$) for 6 weeks. Under the optimum conditions, 46 commercial cultivars of radish were tested for resistance to YC-1 (infecting 15 clubroot-resistant cultivars of Chinese cabbage) and GN-1 (wild type) isolates of P. brassicae. Among them, thirty-five cultivars showed resistance to both isolates and one cultivar represented susceptible response to the pathogens. On the other hand, the other cultivars showed different responses against the tested P. brassicae pathogens. The results suggest that this method is an efficient system for screening radish with resistance to clubroot.

• Research in Plant Disease
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• v.18 no.2
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• pp.86-92
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• 2012

Clubroot caused by Plasmodiophora brassicae Woron. is one of the most important diseases in Brassica crops worldwide. To establish more simple and reliable screening method for resistant cabbage and broccoli to P. brassicae, the development of clubroot on the plants according to inoculum concentration and incubation period after inoculating with the pathogen was investigated using P. brassicae GN1 isolate (race 9). To facilitate and acquire precise result of resistance screening of cabbage and broccoli to clubroot, 14-day-old seedlings were inoculated by drenching roots with the spore suspension of P. brassicae to give inoculum density of $2.5{\times}10^9$ spores/pot. To develop the disease, the inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days, and then cultivated in a greenhouse ($20{\pm}5^{\circ}C$) for five weeks. Under the optimum conditions, 16 cabbage and 17 broccoli cultivars were tested for resistance to four field isolates (GN1, GN2, GS and YC) of P. brassicae collected from four regions in Korea. Among them, some cabbage and broccoli cultivars showed different resistance response to three isolates (GN1, GN2 and GS) determined as race 9 by using the differential varieties of Williams. On the other hand, all the tested cultivars were highly susceptible to YC isolate (race 2). The results suggest that this method is efficient screening method of cabbage and broccoli for resistance to P. brassicae.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.28-28
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• 2015

Clubroot disease caused by Plasmodiophora brassicae induces severe losses of cruciferous vegetables worldwide. To control clubroot of Chinese cabbage, many CR (clubroot resistance) F1 hybrid cultivars have been bred and released in Korea, China and Japan. In this study, we determined the race of P. brassicae 12 field isolates, which collected from 10 regions in Korea, using Williams' differential varieties including two cabbage ('Jersey Queen', 'Badger Shipper') and two rutabaga ('Laurentian', 'Whilhelmsburger'). By Williams' differential varieties, 12 clubroot pathogens were assigned into one (GN2), two (HS and YC), two (HN1 and HN2), three (DJ, KS and SS) and four (GS, GN1, JS and PC) isolates for races 1, 2, 4, 5 and 9, respectively. In addition, the degree of resistance of 45 CR cultivars that were from Korea, China and Japan was tested with the 12 isolates. The 45 CR cultivars of Chinese cabbage were differentiated into three genotypes according to their resistance responses. Even though the 12 P. brassicae isolates were same race by Williams' differential varieties, three CR genotypes showed different resistance response to the isolates. These results indicate that races of P. brassicae by Williams' differentials were not related with resistance of CR cultivars, and three CR genotypes represented qualitative resistance to the P. brassicae isolates. CR genotype I including 'CR-Cheongrok' showed resistance to GN1, GN2, JS, GS, HS, DJ and KS isolates and susceptibility to YC, PC, HN1, HN2 and SS isolates. And CR genotype II such as 'Hangkunjongbyungdaebaekchae' was resistant to GN1, GN2, JS, GS, HS, YC, PC and HN1 and susceptible to DJ, KS, SS and HN2. CR genotype III including 'Chunhajangkun' and 'Akimeki' represented resistance to 10 isolates except for SS and HN2 isolates. Based on these results, we selected 'CR-Cheongrok', 'Hangkunjongbyungdaebaekchae', and 'Chunhajangkun' as a representative cultivar of three CR genotypes and 'Norangkimjang' as a susceptible cultivar. Furthermore, we investigated the resistance of 15 lines of Chinese cabbage, which were provided by seed companies, to 11 isolates except for HN1 of P. brassicae. The results showed that three lines were susceptible to all the tested isolates, whereas five, four, and three lines represented the similar responses corresponding to the CR genotypes I, II, and III, respectively; there is no line of Chinese cabbage showing different resistance patterns compared to three CR genotypes. In particular, line 'SS001' showing resistance responses of CR genotype II was a parent of 'Saerona' that have been commercialized as a CR $F_1$ cultivar of Chinese cabbage. Together, we divided 12 isolates of P. brassicae into 4 races, designated by wild type, mutant type 1, mutant type 2, and mutant type 3. Wild type including GN1, GN2, JS, GS, and HS isolates of P. brassicae was not able to infect all the cultivars of three CR genotypes, whereas, mutant type 3 such as SS and HN2 isolates developed severe clubroot disease on all the CR genotype cultivars. To mutant type 1 including DJ and KS isolates, CR genotypes I, II and III were resistant, susceptible and resistant, respectively. In contrast, to mutant type 2 including YC, PS, and HN1 isolates, CR genotypes I, II and III showed susceptibility, resistance and resistance, respectively. Taken together, our results provide the extended knowledge of classification of P. brassicae races, which is useful information for the breeding of resistant crops, with a suggestion that 'Norangkimjang', 'CR-Cheongrok', 'Saerona' and 'Chunhajangkun' cultivars of Chinese cabbage could be used as new race differentials of P. brassicae for clubroot disease assay.

• Horticultural Science & Technology
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• v.34 no.3
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• pp.442-452
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• 2016

Clubroot caused by Plasmodiophra brassicae Woron. is one of the most important diseases in Brassica crops worldwide. To investigate resistance of cabbage to disease caused by P. brassicae isolates, we evaluated development of clubroot on commercial clubroot resistant (CR) and non-CR cultivars, a CR line, and $F_3$ lines from a cross between a CR line and a non-CR line using several isolates of P. brassicae. Four P. brassicae isolates (DJ, HN1, GN1, and YC) were used to measure development of clubroot on 16 non-CR cabbage cultivars that have been commercialized in Korea. Although four P. brassicae isolates induced similar disease severity on non-CR Chinese cabbage, these isolates exhibited different virulence on the cabbage cultivars. The YC isolate was the most virulent, followed by the GN1, HN1, and DJ isolates. Despite differences in virulence of the isolates on the cabbage cultivars, a CR cabbage line 'YCR478' and two CR cabbage cultivars showed high resistance to 12 P. brassicae isolates including DJ, HN1, GN1, and YC. When three isolates (YC, GN1, and DJ) were inoculated onto 107 $F_3$ lines that were derived from a cross between 'YCR478' and a susceptible cabbage line 'C1176', our results showed that 89, 33, and 6 of $F_3$ lines were susceptible to YC, GN1, and DJ isolates, respectively. In aspects of resistance, 6, 36, and 67 of $F_3$ lines exhibited resistant responses to YC, GN1, and DJ isolates, respectively. Taken together, these results suggest that resistance of cabbage to clubroot is likely affected by the virulence of P. brassicae isolates.

• The Plant Pathology Journal
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• v.18 no.5
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• pp.266-270
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• 2002

Inbred lines of Chinese cabbage KU-101 (resistant line against Plasmodiophora brassicae race race 6) and CS-113 (susceptible line) were crossed and their progeny lines F$_1$, BC$_1$F$_1$, F$_2$, and F$_3$ were produced for the construction of the genetic linkage map of R brassicae race 6-resistant Brassica campestris ssp. pekinensis genome. Restriction fragment length polymorphism (RFLP) was applied to compare between parents and their f$_2$ progenies with a total of 192 probes and 5 restriction enzymes. The constructed RFLP map covered 1,104 cM with a mean distance between genetic marker of 8.0 cM, and produced 10 linkage groups having 121 genetic loci. The loci of P. brassicae race 6 (CR6)-resistant Brassica genome were determined by interval mapping of quan-titative trait loci (QTL), which resulted from bioassay using the same race of the fungi in P3 population. Resistant loci were estimated in numbers 1 (Gl) and 3 (G3) linkage groups. In the regression test, Gl had a value of4.8 logarithm of odd (LOD) score, while C3 had values of 4.2-7.2. Given these results, the location of the CR6-resistant loci within the Brassica genome map can now be addressed.