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1.
A study was conducted to assess the effect of temperature on infection and development of Plasmodiophora brassicae in root hairs of Shanghai pak choi (Brassica rapa subsp. chinensis) and on initiation of clubroot symptoms. Ten‐day‐old seedlings were grown in liquid‐sand culture, inoculated with resting spores and maintained in growth cabinets at 10, 15, 20, 25 and 30°C. Seedlings were harvested at 2‐day intervals, starting 2 days after inoculation (DAI) and continuing until swelling of the tap root was observed (maximum 28 days). Roots were assessed for root hair infection (RHI), stage of development of infection (primary plasmodia, zoosporangia, release of zoospores, secondary plasmodia), symptom development, and for clubroot severity at 24 DAI. Temperature affected every stage of clubroot development; RHI was highest and visual symptoms initiated earliest at 25°C, intermediate at 20 and 30°C, and lowest and latest at 15 and 10°C. Root hair infection was observed at every temperature, but clubroot symptoms developed only above 15°C. A substantial delay in the development of the pathogen was observed at 10 and 15°C. No symptoms were observed at 28 DAI in plants grown at 10°C. Swelling of the tap root was visible at 28 DAI in plants at 15°C, 14 DAI at 20 and 30°C, and 10 DAI at 25°C. These results support and explain the observation in companion studies that cool temperatures result in slower development of clubroot symptoms in brassica crops.  相似文献   

2.
This study investigated the ability of an endophytic fungus Acremonium alternatum to reduce clubroot formation in the model plant Arabidopsis thaliana, which is highly susceptible to Plasmodiophora brassicae . Quantitative PCR demonstrated that A. alternatum colonized the P. brassicae -infected roots and shoots of the host plant. When Arabidopsis plants were co-inoculated with P. brassicae and A. alternatum , gall formation was reduced as shown by the reduction of the disease index (DI) by up to 50% compared to plants only infected with P. brassicae, whereas the infection rate was lowered by about 20% only in several, but not all, experiments. Clubroot was similarly suppressed when plants were inoculated with autoclaved A. alternatum spores or spore extracts, showing that viable spores were not needed. However, A. alternatum spores did not inhibit P. brassicae resting spore germination. Compared to the normal root galls, the smaller root galls on A. alternatum -inoculated plants contained fewer resting spores of the clubroot pathogen. It was thus hypothesized that inoculation with A. alternatum delayed the development of P. brassicae . Using quantitative RT-PCR to monitor the expression of P. brassicae genes differentially expressed during the development of the disease, a delayed pathogen development was corroborated. Furthermore, greenhouse experiments identified a time window in which the endophyte had to be administered, where the latest effective time point was 5 days before inoculation with P. brassicae and the optimum treatment was to administer A. alternatum and P. brassicae at the same time. These results indicate that A. alternatum and perhaps similar endophytes could be useful for the management of clubroot disease.  相似文献   

3.
ABSTRACT Resting spores of Plasmodiophora brassicae were surface-disinfested by treatment with 2% chloramine-T for 20 min and then with an antibiotic solution (1,000 ppm of colistin sulfate, 1,000 ppm of vancomycin hydrochloride, and 6,000 ppm of cefotaxime sodium) for 1 day. The disinfested resting spores were used to inoculate hairy roots of cabbage (Brassica oleracea var. capitata cv. Fuji Wase), Chinese cabbage (B. pekinensis cv. Musou Hakusai), turnip (B. rapa var. rapifera cv. Wase Okabu), and rape (B. napus line Dc 119). Differences among hosts in susceptibility to clubroot in hairy roots were evident. Chinese cabbage and turnip hairy roots supported the highest percentages of root hair infection (53.3 to 80%) and the greatest production of zoosporangial groups (8.5 to 32.5 per root). Moreover, gall formation was observed only on Chinese cabbage and turnip hairy roots. The morphology of zoo-sporangia, plasmodia, and resting spores in diseased hairy roots was found to be identical to that in infected intact plants by both light and scanning electron microscopy. Pathogenicity tests confirmed the infectivity of resting spores produced in hairy roots. Thus, the hairy root culture technique should prove useful as a dual culture system for P. brassicae.  相似文献   

4.
Clubroot of oilseed rape (OSR), caused by Plasmodiophora brassicae, is a disease of increasing economic importance worldwide. Previous studies indicated that OSR volunteers, Brassica crops and weeds play a critical role in the predisposition of the disease. To determine the effect of timing of foliar application of the herbicide glyphosate or mechanical destruction of OSR volunteers in reduction of clubroot severity and resting spore production, a series of studies was conducted under controlled conditions with a susceptible OSR cultivar and an isolate of P. brassicae. Plants were inoculated by injecting a spore suspension beside the root hairs at growth stage 11–12 (BBCH scale) and were terminated at 7 (early) or 21 (late) days post‐inoculation (dpi). Under controlled conditions, the first symptoms on roots were observed as early as 7 dpi. The early application of glyphosate as well as early mechanical destruction resulted in significant (P ≤ 0.05) reduction in the development of clubroot symptoms, root fresh weight and the number of resting spores?g root. Furthermore, the effect of volunteer management on clubroot severity in the succeeding OSR was studied by inoculating plants with the resting spores obtained from treated clubbed roots. Inoculated OSR exhibited root clubs similar to the initial symptoms after 35 dpi. Plants that were inoculated with spore suspension from early treated roots resulted in significant reductions in clubroot incidence and severity. Conversely, plants inoculated with the spore suspension from the late treated roots displayed levels of clubroot similar to the plants inoculated with the spore solutions of positive controls.  相似文献   

5.
To investigate the susceptibility of hairy root lines of Brassica species to Plasmodiophora brassicae, hairy roots were induced in a number of Brassica species with Agrobacterium rhizogenes. Turnip hairy root was highly susceptible to P. brassicae; infection rates were high and large galls formed. In contrast, the rates of root hair infection and gall formation on intact Brassica plants did not differ significantly from the control. To induce resting spore formation, turnip hairy roots were incubated at 15°, 20°, or 25°C after 3 weeks of incubation at 25°C. The number and fresh mass of the galls per hairy root were higher and formation of resting spores was greatest after a 7-week incubation at 20°C. To subculture P. brassicae using turnip hairy root, turnip hairy roots were reinoculated with resting spores and gall with resting spores then formed on the hairy roots. In this way, P. brassicae using hairy roots could be subcultured in vitro two or three times on three single-spore isolates of P. brassicae. This is the first report of in vitro subculture of P. brassicae using hairy root.  相似文献   

6.
Clubroot of crucifers, caused by the obligate parasite Plasmodiophora brassicae, is characterized by the formation of conspicuous root galls. These galls usually have a club- or spindle-shaped morphology, and interfere with water and nutrient uptake by infected plants. Smaller galls, historically regarded as resistance structures and distinct from the typical spindle-shaped galls, have also been identified and termed ‘spheroid galls’ because of their spherical or nearly spherical form. An assessment of various Brassica species and varieties revealed that spheroid galling could be observed in all genotypes investigated, but occurred regularly only in a few particular host/P. brassicae combinations. While spindle gall formation was coincident with the expansion of the stele and infection of secondary tissues by P. brassicae, spheroid galls typically had a region of proliferating tissue that corresponded to the secondary cortex and periderm of the healthy plants, with the outer proliferating tissue less infected than the inner portions. The underlying host tissue showed limited secondary tissue development, was largely uninfected, and, where infection occurred, a continuous stele was maintained. An active host defensive reaction, in the form of cell lignification or the hypersensitive response, was not observed, while pathogen resting spores were visible in one longitudinal section of a spheroid gall. These findings suggest that while the proliferation of P. brassicae is restricted in spheroid galls, these structures are not indicative of complete resistance to clubroot.  相似文献   

7.
A resistant type of small, spheroid clubroot galls (SSGs) containing resting spores formed on the root surface of clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa L. subsp. pekinensis) inoculated with an avirulent population of Plasmodiophora  brassicae. Populations isolated from the SSGs severely affected a common (susceptible) cultivar but did not have the typical pathogenicity on CR cultivars, indicating similarity in pathogenicity between SSG and original spore populaions. Populations pathogenic on CR cultivars were not detected from SSGs. Therefore, the ability of the avirulent population among the SSGs to form resting spores may need to be considered to achieve clubroot control in common cultivars of crucifers.  相似文献   

8.
The pathogenesis of clubroot, a disease of cruciferous crops caused by the fungusPlasmodiophora brassicae, starts with infection of the root hairs. This process was studied in 13 accessions ofBrassica oleracea, B. napus and B. rapa with varying levels of plant resitance toP. brassicae. Seedlings were grown in a mineral solution, inoculated with resting spores ofP. brassicae, and the number of plasmodia developing in root hairs was recorded. When compared with the standard susceptible cultivar Septa, both higher and lower resistance to root hair infection was found in the accessions of the differentBrassica species. No complete resistance to root hair infection was found. Over the accessions studied, there was no correlation between the plant resistance estimated from greenhouse tests and the resistance to root hair infection of seedlings. The resistance of all accessions must at least partly be caused by other mechanisms which operate after the root hair plasmodia are formed.  相似文献   

9.
The expression of 12 cDNAs from Plasmodiophora brassicae , among them two novel sequences, was determined during clubroot development on Arabidopsis thaliana . The aim was to find cDNAs expressed at distinct stages of pathogenesis. The relative amount of infection with active plasmodia could be estimated using Pb Actin cDNA as an internal standard. Two cDNAs, Pb Brip9 and Pb CC249, were strongly expressed at stages of disease development corresponding to the occurrence of sporulating plasmodia. Therefore, it should be possible in the future to find more cDNAs which could be used as markers for certain stages of clubroot development.  相似文献   

10.
The mechanism of the biofungicide Prestop® (Clonostachys rosea) was investigated for control of clubroot (Plasmodiophora brassicae) on canola. The key product components were partitioned and assessed for their effect on pathogen resting spores, root hair infection (RHI) and disease development using light microscopy, quantitative PCR and different application treatments during infection. The whole product of Prestop was consistently more effective than the C. rosea conidial suspension or product filtrate alone in reducing RHI and clubroot development. This biofungicide showed little effect on germination or viability of resting spores. Two‐application treatments at seeding and 7–14 days after seeding achieved greater clubroot control than a single application of the biofungicide at either seeding or post‐seeding stage. This may indicate the need to maintain a high biofungicide dose in the soil during primary and secondary infection. This biocontrol fungus colonized the rhizosphere and interior of canola roots extensively, and possibly induced plant resistance based on up‐regulation of the genes that are involved in jasmonic acid (BnOPR2), ethylene (BnACO) and phenylpropanoid (BnOPCL, BnCCR) biosynthetic pathways. It is concluded that the biofungicide Prestop suppressed clubroot on canola at least via root colonization and induced systemic resistance (ISR), and the latter may be through the modulation of phenylpropanoid and jasmonic acid/ethylene metabolic pathways elicited by the fungus.  相似文献   

11.
Clubroot, a disease of Brassicaceae species, is caused by the soilborne pathogen Plasmodiophora brassicae. High soil water content was previously described to favour the motility of zoospores and their penetration into root cells. In this study, the effect of irrigation regimes on clubroot development during the post‐invasive secondary phase of infection was investigated. Three irrigation regimes (low, standard, high) were tested on two Arabidopsis accessions, Col‐0 (susceptible) and Bur‐0, a partially resistant line. In Col‐0, clubroot symptoms and resting spore content were higher under the ‘low irrigation’ regime than the other two regimes, thus enhancing the phenotypic contrast between the two Arabidopsis accessions. Clubroot severity under high and low irrigation regimes was evaluated in near‐isogenic lines derived from a Col‐0 ×  Bur‐0 cross, to assess the effect of soil moisture on the expression of each of four quantitative trait loci (QTL) controlling partial resistance. The presence of the Bur‐0 allele at the QTL PbAt5.2 resulted in reduced severity only under low irrigation, whereas the Bur‐0 allele at QTL PbAt5.1 was associated with partial resistance only under high irrigation. QTL PbAt4 reduced the number of resting spores in infected roots, but was not associated with reduced clubroot symptoms. The results indicated that soil moisture could have consequences for the secondary phase of clubroot development, depending on plant genotype. Future genetic studies may benefit from using combinations of watering conditions during the secondary stage of infection, thus opening up the possibility of identifying genetic factors expressed under specific environmental conditions.  相似文献   

12.
U. MERZ 《Plant pathology》1997,46(5):670-674
A solution culture test system with Spongospora subterranea f.sp. subterranea spore ball inoculum and tomato bait plants was used to create a pulse of primary zoospore production and subsequent host-root infection. Spore balls and zoospores were examined by light, fluorescence, and transmission and scanning electron microscopy. Most of the resting spores with a developing exit pore did not show any changes in cytoplasmic content typical of zoospore formation. A few empty resting spores and some with developing exit pores were also observed in the absence of host-root exudates. The average diameter of exit pores of empty resting spores was 1.5 μm and they were often encircled by a ring-like fusion of wall layers. Mature zoospores were never found inside resting spores. Primary and secondary zoospores are identical in morphology. The infection process is similar to that of other Plasmodiophoromycetes with internal 'Rohr'-like structures in encysted zoospores which were attached by an adhesorium to tomato root hairs. Post-infection papillae and uninucleate plasmodia were observed.  相似文献   

13.
Using quantitative PCR, DNA of Plasmodiophora brassicae, the causal agent of clubroot, was detected and quantified on canola, pea and wheat seeds, as well as on potato tubers, all harvested from clubroot‐infested fields in Alberta, Canada. Quantifiable levels of infestation were found on seven of the 46 samples analysed, and ranged from <1·0 × 103 to 3·4 × 104 resting spores per 10 g seeds; the vast majority (80–100%) of resting spores on these samples were viable, as determined by Evan’s blue vital staining. However, the levels of infestation found were generally lower than that required to cause consistent clubroot symptoms in greenhouse plant bioassays. While the occurrence of P. brassicae resting spores on seeds and tubers harvested from clubroot‐infested fields suggests that seedborne dissemination of this pathogen is possible, practices such as commercial seed cleaning may be sufficient to effectively mitigate this risk.  相似文献   

14.
Controlled‐environment studies were conducted on two Brassica crops (canola, Brassica napus; and Shanghai pak choi, B. rapa subsp. chinensis var. communis) to examine the effects of temperature on infection and subsequent development of clubroot caused by Plasmodiophora brassicae. In the first experiment, canola seedlings were grown in infested soil for 3 weeks at 14–26°C to assess the impact on primary and secondary infection and transferred to 20°C for 3 weeks to assess symptom development under uniform conditions, or started at 20°C for 3 weeks and then placed at the treatment temperatures for the final 3 weeks to assess the impact of temperature on symptom development. A second experiment examined a wider range of temperatures (10–30°C). Similar experiments were also conducted on Shanghai pak choi. The studies demonstrated that clubroot severity was affected by temperature during both infection and vegetative development of the crop. Both early and late in crop development, little or no clubroot developed at temperatures at or below 17°C, and development was slower above 26°C than at 23–26°C for both crops throughout the study. In canola, the high levels of inoculum used in the study resulted in a high incidence of clubroot irrespective of temperature, but in pak choi incidence showed the same pattern as severity. This is the first study to demonstrate under controlled conditions that temperature during vegetative growth of the crop affects symptom development of clubroot.  相似文献   

15.
Clubroot, caused by Plasmodiophora brassicae, has emerged as a serious disease threatening cruciferous crop production throughout the world. Crop rotation with non-host species is commonly practised to avoid clubroot, but it is not known whether rotation crops can control clubroot when the resting spores of P. brassicae remain unaffected. Pot experiments were performed to investigate the response of clubroot in Chinese cabbage to crop rotation with potato onion. The results showed that Chinese cabbage rotated with potato onion exhibited less clubroot disease than Chinese cabbage monoculture. Compared with residues from potato onion, the addition of root exudates from potato onion significantly decreased the disease incidence and index of clubroot (p ≤ 0.05). Potato onion root exudates decreased the number of secondary plasmodia of P. brassicae and the expression of the PRO1 gene of P. brassicae. These results suggest that root exudates from potato onion may play an important role in suppressing clubroot in a Chinese cabbage-potato onion-Chinese cabbage rotation system.  相似文献   

16.
Control of some soilborne pathogens may be achieved by use of decoy or catch crops. These stimulate the germination of resting spores, resulting in limited expression of disease symptoms. Results achieved using this approach are reported here using leafy daikon (radish, Raphanus sativus var. longipinnatus ) for control of Plasmodiophora brassicae , the cause of clubroot disease of Brassicaceae. Disease indices of Chinese cabbage plants grown in pots that had previously contained leafy daikon were lower compared with pots where no plants had been grown before (control pots). Numbers of resting spores of P. brassicae in soil in pots after cultivation with leafy daikon were reduced by 71% compared with control pots when resting spores were recovered and counted directly. In a field experiment, numbers of resting spores were reduced by 94% compared with the start of the experiment when leafy daikon was grown in advance of Chinese cabbage, but there was no reduction in disease severity in the Chinese cabbage. Plasmodiophora brassicae infected the root hairs of leafy daikon and those of Chinese cabbage, but no clubs were found on leafy daikon roots. The results from pot trials indicate that leafy daikon may be useful as a decoy crop for the control of clubroot disease in field crops.  相似文献   

17.
To mitigate the impact and dissemination of clubroot in western Canada, canola (Brassica napus) producers have relied on clubroot resistance traits. However, in 2013 and 2014, new strains of the clubroot pathogen, Plasmodiophora brassicae, emerged that are virulent on most clubroot‐resistant (CR) canola genotypes. Novel strains of the pathogen were inoculated onto two susceptible canola cultivars, one resistant line and six CR cultivars. Although all cultivars/lines showed a susceptible response to inoculation with the new strains of P. brassicae, the severity of disease reaction, root hair infection rates and the amount of P. brassicae DNA present in each canola genotype varied depending on the strain. In addition, the effect of inoculum density on disease severity and gall formation was recorded for one of these new strains on a universally susceptible Chinese cabbage cultivar and one susceptible and 10 resistant canola genotypes. Although root galls were observed at an inoculum density of 103 spores per mL of soil, clear differentiation of susceptible and resistant reactions among canola cultivars/lines was not observed until the inoculum density reached 105 spores mL?1. At a spore density of 106 spores mL?1 and above, all cultivars/lines developed susceptible reactions, although there was some differentiation in the degree of reaction. This study shows the potential to develop a unique disease profile for emergent clubroot pathotypes and shows a useful range of spore densities at which to study new P. brassicae strains.  相似文献   

18.
The early stages of infection of canola roots by the clubroot pathogen Plasmodiophora brassicae were investigated. Inoculation with 1 × 105 resting spores mL?1 resulted in primary (root hair) infection at 12 h after inoculation (hai). Secondary (cortical) infection began to be observed at 72 hai. When inoculated onto plants at a concentration of 1 × 104 mL?1, secondary zoospores produced primary infections similar to those obtained with resting spores at a concentration of 1 × 105 mL?1. Secondary zoospores caused secondary infections earlier than resting spores. When the plants were inoculated with 1 × 107 resting spores mL?1, 2 days after being challenged with 1 × 104 or 1 × 105 resting spores mL?1, secondary infections were observed on the very next day, which was earlier than the secondary infections resulting from inoculation with 1 × 107 resting spores mL?1 alone and more severe than those produced by inoculation with 1 × 104 or 1 × 105 resting spores mL?1 alone. Compared with the single inoculations, secondary infections on plants that had received both inoculations remained at higher levels throughout a 7‐day time course. These data indicate that primary zoospores can directly cause secondary infection when the host is under primary infection, helping to understand the relationship and relative importance of the two infection stages of P. brassicae.  相似文献   

19.
Infectivity of resting spores ofPolymyxa betae in soil stored air-dry or moist was determined by assessing infection of bait plants that were exposed to the soil. Storage of soil under air-dry conditions at room temperature resulted in a delayed onset of germination of resting spores compared to germination in soil stored under moist and cool conditions, as inferred from the infection of the bait plants. Bait plants had to be exposed for more than 12 h to flooded infested soil before germination and infection had occurred. However, when soil was prewetted for 24 h before exposing bait plants, germination, infection and transmission of beet necrotic yellow vein virus (BNYVV) were accomplished within 12 h, but only with the moistly stored soil. When resting spores isolated from roots were stored for 4 and 8 weeks under dry conditions at 22°C, germination of viruliferous spores, as measured by detection of BNYVV in bait plants exposed for 48 h to the spores, was less than that of spores stored in moist soil at 22°C. Approximately 100% of bait plants were infected after exposure to resting spores that were frozen in demineralized water or stored cool (5°C) in water or moist soil for 42 weeks. Air-dry cool storage for 42 weeks resulted in a low percentage of infection. Storage conditions of soil influence the results of bioassays for detection of rhizomania when short baiting periods are applied, whereas differences in infectivity were not detected using a bioassay with long duration.  相似文献   

20.
Clubroot, caused by Plasmodiophora brassicae, is a worldwide disease affecting Brassica. Until now, the detection of genetic factors (QTLs) implicated in clubroot resistance has been based on estimates of disease index. However, as the amount of resting spores released in soil after club disintegration influences clubroot epidemics and resistance‐breaking dynamics, its genetic control may deserve specific attention. In a previous report, it was shown that nitrogen fertilization modulated quantitative partial resistance toward clubroot symptom development in rapeseed. The present work aimed to identify genetic factors involved in the control of resting spore production and to assess their regulation by nitrogen supply. A flow cytometer method was adapted for rapidly estimating resting spore content in a large series of samples. Linkage analysis was conducted to detect QTLs implicated in resting spore production in a Brassica napus doubled haploid progeny from the cross Darmor‐bzh × Yudal. DH lines inoculated with the P. brassicae isolate eH were grown under low‐ and high‐nitrogen supply. Under low‐nitrogen conditions, resting spore production was reduced compared to high‐nitrogen conditions, regardless of genotypes. Genetic architecture controlling resting spore production and clubroot symptom development was similar. Under high‐nitrogen conditions, resting spore production was controlled by one major QTL (C09a) and a few small‐effect QTLs. By contrast, two major QTLs (C02 and C09a) controlled resting spore production under low‐nitrogen conditions. This work highlighted a large see‐saw effect between the relative contribution of the C09a QTL (high effect under high‐nitrogen conditions) and the C02 QTL (high effect under low‐nitrogen conditions), with possible implications in resistance breeding.  相似文献   

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