Reduction of Spore Density of Plasmodiophora brassicae in Soil by Decoy Plants

The severity of clubroot (Plasmodiophora brassicae) on Chinese cabbage was reduced by growing plants such as oats, spinach and leafy daikon prior to Chinese cabbage in pot experiments. Resting spore densities of P. brassicae in the soil were 29–62%, depending on the pervious crop, as compared to unplanted control plot after ploughing under the previously cultivated plants. Root hairs of the preceding plants were infected with P. brassicae, but clubbed roots were not formed on these plants. The results indicate that these plants functioned as decoy plants reducing the resting-spore density in soil and thereby suppressing disease severity. Received 21 February 2000/ Accepted in revised form 5 September 2000     

A molecular marker for the specific detection of new pathotype 5‐like strains of Plasmodiophora brassicae in canola

Clubroot of crucifers, caused by Plasmodiophora brassicae, is managed in canola (Brassica napus) by the deployment of resistant cultivars. Recently, however, new strains of P. brassicae have been detected in Alberta, Canada, that can overcome this resistance. Some of these strains are classified as pathotype 5 on the differential system of Williams, but are distinguished by their ability to overcome host resistance. In order to expedite the identification of these new pathotype 5‐like strains, three primer sets were developed based on the 18S‐ITS region of the pathogen. With primers P5XF3 and P5XR3, a 127 bp product was amplified from all new pathotype 5‐like strains following optimized PCR analysis. A TaqMan probe‐based quantitative assay was also developed. These protocols could be used to detect as little as 0.5 pg P. brassicae DNA, and as few as 10⁴ mL^?1 pathogen resting spores; infection of host tissues could be detected as soon as 4 days after inoculation. The PCR and qPCR assays described in this study represent useful tools for the rapid and reliable diagnosis and quantification of new pathotype 5‐like strains of P. brassicae.     

Suppression of the clubroot pathogen Plasmodiophora brassicae by plant growth promoting formulations in roots of two Brassica species

In this study, the ability of two organic plant growth stimulants, mainly based on algal extracts, amino acids and phosphonate, to reduce clubroot formation in the model plant Arabidopsis thaliana and two economically important Brassica species, Brassia rapa (Chinese cabbage) and Brassica napus (oilseed rape) was investigated. A commercial liquid (Frutogard^®) and a granulate (PlasmaSoil^®) formulation were used to find optimum conditions for both control of the pathogen and plant growth. Both formulations were able to significantly reduce gall formation after Plasmodiophora brassicae infection on Chinese cabbage, but PlasmaSoil^® gave better effects, possibly as a result of the continuous supply of the components to the soil. Individual components did not have the same effect. Clubroots on oilseed rape could also be reduced. In contrast, club formation was not reduced in the model plant Arabidopsis thaliana. This points to interesting differences in the induction of resistance in the different species. Cross‐sections of infected and treated roots stained for different macromolecules (callose, lignin, suberin) indicated differences in anatomy as a result of the two formulations. The results indicate an application for the granulate formulation PlasmaSoil^® in clubroot control.     

Induction of systemic resistance to Sclerotiniasclerotiorum by oxalic acid in oilseed rape

Treatment of the lowermost leaf of flowering plants of oilseed rape cv. Linetta with 20 m m oxalic acid induced systemic disease resistance in the three next youngest leaves. Resistance was not caused by the translocation of fungitoxic concentrations of oxalic acid. Resistance was also expressed in the stem; in oxalic acid-treated plants the vertical spread of stem lesions was halted and the signal travelled both acropetally and basipetally. Expression of resistance in the leaves was detected 6 h after treatment and continued for at least 5 weeks. Twenty-four h after treatment, resistance was detectable in the leaves following the use of as little as 0.5 m m oxalic acid. Local resistance in the oxalic acid-sprayed leaf was detectable only at certain times (12 and 48 but not 24 h after treatment) and at very low (0.25 m m ) or high (≥40 m m ) oxalic acid concentrations. When oxalic acid was applied to a discrete area of the leaf, significant local resistance was expressed in the surrounding leaf tissue, maximum resistance being exhibited by the tissue closest to the site of petiole attachment.     

江西油菜根肿菌生理小种鉴定及品种抗病性鉴定

油菜根肿病在江西省多市均有分布,尤以上饶婺源、吉安永新、九江瑞昌等地发生严重。为明确江西省油菜主产区根肿菌的生理小种,采用SCD鉴定系统对江西油菜主产区根肿菌进行了生理小种鉴定,结果共鉴定出3个生理小种类型：Pb1、Pb5和Pb12。为筛选出适于江西根肿病发病地区种植的油菜材料,开展了油菜材料对根肿菌的抗性测定。室内盆栽抗性测定表明,育种中间材料‘P360429001’‘2018362481’‘P360924009’‘P360281040’‘中油杂12’‘华油杂62R’和‘华双5R’的病情指数均低于或等于30.00,其中‘华油杂62R’和‘华双5R’表现为中抗水平。田间抗性测定结果表明,‘华油杂62R’‘秦优DK4’‘阳光50’和育种中间材料‘P360429001’‘ZS115R’的根肿病病情指数均低于30.00,但只有‘华油杂62R’对Pb1生理小种表现出中抗水平。本研究针对性地筛选出适于江西油菜根肿病发病地区种植的油菜材料,对江西油菜根肿病的防控和创制抗病性油菜资源提供了科学依据。     

影响油菜根肿病发生流行的因子分析

近年来,十字花科根肿病在安徽黄山油菜种植区发生日趋严重,成为限制油菜产量和品质的重要因素。本文通过大田调查、田间试验并结合气象资料分析,表明品种抗性、土壤性质及种植方式、前茬作物和气象因子等是影响该病发生的主要因素。不同油菜品种根肿病的发病情况存在显著性差异。‘汇油50’最感病,‘浙双72’最抗病。移栽田较直播田发病轻。砂壤土油菜根肿病发生最重,黏性土发病次之,壤土发病最轻,前茬为甘薯、水稻和玉米的田块,发病轻,前茬为十字花科蔬菜的田块,油菜根肿病发生最重,前茬为大豆的田块,发病次之。9月份月均温、9月上旬降雨量和9月下旬降雨量等气象因子与根肿病发病率关系最密切。     

Influence of cultivar resistance and inoculum density on root hair infection of canola (Brassica napus) by Plasmodiophora brassicae

The impact of cultivar resistance and inoculum density on the incidence of primary infection of canola root hairs by Plasmodiophora brassicae, the causal agent of clubroot, was assessed by microscopy. The incidence of root hair infection in both a resistant and a susceptible cultivar increased with increasing inoculum density, but was two‐ to threefold higher in the susceptible cultivar; the relationship between root hair infection and inoculum density was also substantially stronger and more consistent in the susceptible cultivar. In the susceptible cultivar, the root hair infection rate peaked between 6 and 8 days after sowing and then declined. In the resistant cultivar, it increased over the 14‐day duration of each study. It appears that examination of root hair infection by microscopy in a bait crop of susceptible canola could serve as a useful tool for estimating P. brassicae inoculum levels in soil. In a separate trial, the relationship between inoculum density and clubroot severity, plant growth parameters, and seed yield was assessed under greenhouse conditions. Inoculum density in the susceptible genotype was strongly and positively correlated with clubroot severity and negatively correlated with plant height and seed yield. In addition, a single cropping cycle of the susceptible cultivar contributed significantly higher levels of resting spores to the soil in a greenhouse test than did a cycle of the resistant cultivar, as assessed by quantitative PCR and microscope analysis.     

Interactions in the Brassica napus–Pyrenopeziza brassicae pathosystem and sources of resistance to P. brassicae (light leaf spot)

Pyrenopeziza brassicae, cause of light leaf spot (LLS), is an important pathogen of oilseed rape and vegetable brassicas and has a wide geographic distribution. Exploitation of host resistance remains the most sustainable and economically viable solution for disease management. This study evaluated 18 oilseed rape cultivars or breeding lines for host resistance against P. brassicae in glasshouse experiments. Selected cultivars/lines were inoculated with eight single-spore isolates of the pathogen obtained from three different regions in England. Analysis of P. brassicae infection-related changes on host plants identified leaf deformation as a characteristic feature associated with P. brassicae infection, this showed poor correlation to LLS severity measured as the amount of pathogen sporulation on infected plants. Resistant host phenotypes were identified by limitation of P. brassicae sporulation, with or without the presence of a necrotic response (black flecking phenotype). Investigation of this pathosystem revealed significant differences between cultivars/lines, between isolates, and significant cultivar/line-by-isolate interactions. In total, 37 resistant and 16 moderately resistant interactions were identified from 144 cultivar/line-by-isolate interactions using statistical methods. Most of the resistant/moderately resistant interactions identified in this study appeared to be nonspecific towards the isolates tested. Our results suggested the presence of isolate-specific resistant interactions for some cultivars. Several sources of resistance have been identified that are valuable for oilseed rape breeding programmes.     

Prediction of competition between oilseed rape and Stellaria media

Ten experiments have investigated competition between winter oilseed rape and Stellaria media (common chickweed). Yield losses caused by this weed were often high, but differed greatly between experiments, 5% yield loss being calculated to be caused by 1.4–328 plants m^?2. Predictions of yield loss based on relative weed dry weights [weed dry weights/(crop + weed dry weights)] in December were somewhat less variable than those based on weed density, 5% yield loss being caused by 1.4–10.6% relative weed dry weight. The variations in yield loss were related to variations in the competitiveness of the oilseed rape and the S. media, caused by weather differences between years and sites, and the long period between weed assessment and harvest (8–10 months). However, despite the lack of precise relationships, there were indications that the greater the crop dry weights in December, the lower the final yield loss. Delayed sowing of oilseed rape until late September did not clearly increase the competitive effects of the weed compared with late August/early September sowings. Weed competition was not clearly affected by reduced crop density (44–113 plants m^?2), because of the compensatory ability of the lowest density. The results of the experiments are discussed in relation to the prediction of yield loss and, thus, possible adjustment of weed control strategies to meet expected crop losses.     

Sporulation of Pyrenopeziza brassicae (light leaf spot) on oilseed rape (Brassica napus) leaves inoculated with ascospores or conidia at different temperatures and wetness durations

In controlled environment experiments, sporulation of Pyrenopeziza brassicae was observed on leaves of oilseed rape inoculated with ascospores or conidia at temperatures from 8 to 20°C at all leaf wetness durations from 6 to 72 h, except after 6 h leaf wetness duration at 8°C. The shortest times from inoculation to first observed sporulation ( l ₀), for both ascospore and conidial inoculum, were 11–12 days at 16°C after 48 h wetness duration. For both ascospore and conidial inoculum (48 h wetness duration), the number of conidia produced per cm² leaf area with sporulation was seven to eight times less at 20°C than at 8, 12 or 16°C. Values of Gompertz parameters c (maximum percentage leaf area with sporulation), r (maximum rate of increase in percentage leaf area with sporulation) and l ₃₇ (days from inoculation to 37% of maximum sporulation), estimated by fitting the equation to the observed data, were linearly related to values predicted by inserting temperature and wetness duration treatment values into existing equations. The observed data were fitted better by logistic equations than by Gompertz equations (which overestimated at low temperatures). For both ascospore and conidial inoculum, the latent period derived from the logistic equation (days from inoculation to 50% of maximum sporulation, l ₅₀) of P. brassicae was generally shortest at 16°C, and increased as temperature increased to 20°C or decreased to 8°C. Minimum numbers of spores needed to produce sporulation on leaves were ≈25 ascospores per leaf and ≈700 conidia per leaf, at 16°C after 48 h leaf wetness duration.     

湖北和安徽省油菜病毒病调查和病毒血清鉴定

2003年秋至2005年春2个油菜生长季分别到湖北和安徽2省12县市油菜产区进行病毒病发病情况调查,除局部地区和地块发病率达到20%～40%,多数大面积油菜发病率在0.1%以下。应用间接酶联免疫吸附法检测258份病害样品,TuMV占样品总数的90.7%,CMV占样品总数的8.9%,ORMV占样品总数的0.8%。     

盾壳霉控制油菜菌核病菌再侵染及其叶面存活动态的研究

 本文评估了施于油菜(Brassica napus)叶片上的盾壳霉(Coniothyrium minitans)控制油菜菌核病菌再侵染能力,探讨了其作用机理,并测定了盾壳霉分生孢子在油菜叶面上的存活动态。结果如下:叶面上的盾壳霉对油菜菌核病菌的初侵染影响较小,但在高剂量(> 10⁶孢子/ml)时可以控制病斑的扩展。所有供试剂量的盾壳霉均可不同程度地控制再侵染。盾壳霉分生孢子可在叶面病部迅速萌发,48 h和72 h时孢子萌发率分别为51%和95%,而在健康叶面上6 d未能检测到萌发的孢子。自携带盾壳霉的叶面病部不能分离到核盘菌,表明叶面上的盾壳霉已寄生并破坏了核盘菌再侵染菌丝。自油菜叶面上分离到的盾壳霉菌落数随时间延长而降低,但其分生孢子至少可以在叶面上存活28 d。这即表明,在叶面上适时适量地添加盾壳霉可以控制油菜菌核病的为害。     

Direct evidence of surface infestation of seeds and tubers by Plasmodiophora brassicae and quantification of spore loads

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.