Factors affecting the pattern of the dose response curve of clubroot disease caused by Plasmodiophora brassicae

The severity of clubroot disease caused by Plasmodiophora brassicae Woron. depended on the density of resting spores in soil, soil type, soil pH, and host susceptibility. The dose response curve (DRC) was determined to indicate the relationship between the disease index and these parameters for predicting the disease severity for each field. DRC patterns varied with the soil types and even among soils of the same type depending on the sampling areas. Disease incidence was lower in the soils adjusted to a higher pH than in those at the initial pH. DRC patterns were also influenced by plant species or cultivars. The DRC is useful for estimating the potential disease severity in agricultural fields with different soils and different plants and essential for the development of an integrated pest management (IPM) strategy for the control of clubroot disease.     

芸薹根肿菌SSR标记的开发及遗传多样性研究

芸薹根肿菌（Plasmodiophora brassicae）专性寄生于十字花科作物根部引起根肿病,且变异迅速。本研究利用Williams鉴别寄主系统鉴定出32株根肿菌,根据根肿菌基因组序列分析了根肿菌基因组中的简单重复序列（Simple sequence repeats,SSR）的分布特性,并结合SSR标记的开发研究了根肿菌遗传多样性。主要结果如下：从收集自8个省市31个地区的32株根肿菌中,鉴定出2、4、7和11号生理小种;根肿菌基因组存在3 164个SSR,即每7.6 kb存在1个SSR。核苷酸重复类型以2和3为主,分布频率分别为45.92％和53.04％。从62对SSR引物中,筛选出31个多态性标记,多态率50％,平均PIC值为0.407;聚类分析表明,同一地区的根肿菌大多聚类到一起,但同时存在变异。     

十字花科根肿病菌(Plasmodiophora brassicae)致病力分化研究方法

目前国际上已有Williams、ECD、Somé和Kuginuki 4套鉴别系统用于十字花科作物根肿病菌(Plasmo-diophora brassicae Woron)生理小种和致病型的鉴定。其中Williams和ECD鉴别系统被广泛应用,但还存在一些不足。Somé和Kuginuki分别根据法国和日本的病原菌致病力分化特点,在Williams和ECD鉴别系统的基础上,建立Somé和Kuginuki两个鉴别系统。根肿病菌的单孢子分离是生理小种或致病型鉴定中的研究难点,目前主要采用琼脂法和液滴法。不同研究人员使用的病害鉴定方法及分级标准不同,目前主要采用蘸根法、点滴法和菌土法。     

3株放线菌对白菜根肿病的防治效果及机制研究

研究3株放线菌对白菜根肿病的防治效果,以挖掘具有良好防治效果的生防菌株。以前期分离筛选的2株优良生防放线菌株(Streptomyces pactum Act12和Sr.ochei D74)与1株分离自根肿病发病小区健株根际土中的黄白链霉菌(S.alboflavus SP12)为研究对象,利用孢子萌发、皿内发芽及盆栽试验研究不同菌株对白菜根肿病的防治效果及机制。结果表明:(1)SP12与D74接种可显著降低根肿病的发病率与病情指数,发病率分别降低50.0%和41.7%,病情指数分别降低20.3%和19.4%;SP12与D74提高白菜地下部分鲜质量36.6%和8.6%,同时地上部分鲜质量分别增加25.4%和4.9%。(2)SP12与D74显著影响白菜生长及抗性相关生理生化指标:SP12与D74使根系活力分别增加46.2%和34.4%,PPO活力分别增加49.0%和66.5%,脯氨酸含量分别增加11.2%和35.1%,丙二醛含量分别降低5.2%和7.4%;SP12处理可提高POD活性26.4%(P0.05)。(3)SP12和D74可改变白菜根际土壤微生物群落结构组成,提高放线菌的数量63.7%和15.4%。结果表明:SP12、D74对根肿病防治、植株生长、抗逆酶系统及根际微生态均有良好的促进作用。     

Colonization by two isolates of Plasmodiophora brassicae with differing pathogenicity on a clubroot-resistant cultivar of Chinese cabbage (Brassica rapa L. subsp. pekinensis)

A commercial clubroot-resistant F₁ cultivar of Chinese cabbage (Brassica rapa L. subsp. pekinensis), Kukai 70, is resistant to an isolate of populations of Plasmodiophora brassicae from Hagi (HG) city and is susceptible to another from Yamaguchi (YMG) city. The degree and frequency of primary and secondary cycle colonization by the isolates in the root hairs and root tissues of cv. Kukai 70 were compared. Seedlings of cv. Kukai 70 were grown in soils amended with inoculum of either HG or YMG and harvested 10 days after inoculation to observe the primary cycle (number of root-hair infections) and 20, 30, and 40 days after inoculation to observe of the secondary cycle (frequency of infected cells and degree of plasmodial development based on the number of nuclei in infected cells). Although more root hairs were infected in HG than in YMG, fewer cells in root tissues including the cortex and medullary rays were infected in HG than in YMG. In addition, YMG developed plasmodia with many nuclei and formed resting spores, whereas plasmodia remained immature with a small number of nuclei in HG and did not form resting spores even by 40 days after inoculation. These results suggest that suppression of plasmodial development during secondary colonization is associated with resistance mechanisms to HG in cv. Kukai 70. Starch did not accumulate (i.e., development of amyloplasts) in HG-infected cells. This may be involved in the suppression of secondary colonization of P. brassicae in the cultivar.     

两株根肿病生防放线菌的鉴定及其防病效果

 ：【目的】鉴定从白菜根际土壤及红豆树根部分离得到的抑制根肿病菌休眠孢子萌发的生防菌株A316和A10,并探明其防病潜能,对根肿病进行有效的生物防治。【方法】根据菌株A316和A10的形态与培养特征、生理生化特性、16S rDNA序列对其进行鉴定;评价菌株A316和A10在室内盆栽及大田小区试验中对白菜根肿病的防治效果。【结果】菌株A316与链霉菌中的灰红链霉菌（Streptomyces griseoruber） NBRC 12873亲缘关系最近,同源性达99.9%,且形态与培养特征、生理生化特性与灰红链霉菌相符。菌株A10与GenBank数据库中多个相似性序列的亲缘关系都较近,鉴于其形态与培养特征、生理生化特性,只能归于链霉菌中的灰褐类群。菌株A316、A10对白菜根肿病的室内盆栽防效分别为72.8%、67.2%,而大田小区试验中防效依次达68.5%、56.7%,且可分别使白菜单位面积产量增加96.1%、64.9%。【结论】菌株A316鉴定为灰红链霉菌;菌株A10归于链霉菌中的灰褐类群（S. sp.）。菌株A316和A10对芸苔根肿菌显示出极好的生防潜能,A316的防效好于A10。     

Increase in soil pH due to Ca-rich organic matter application causes suppression of the clubroot disease of crucifers

Clubroot disease of cruciferous plants caused by the soil-borne pathogen Plasmodiophora brassicae is difficult to control because the pathogen survives for a long time in soil as resting spores. Disease-suppressive and conducive soils were found during the long-term experiment on the impact of organic matter application to arable fields and have been studied to clarify the biotic and abiotic factors involved in the disease suppression. The fact that a large amount of organic matter, 400 t ha⁻¹ yr⁻¹ farmyard manure (FYM) or 100 t ha⁻¹ yr⁻¹ food factory sludge compost (FSC), had been incorporated for more than 15 yr in the suppressive soils and these soils showed higher pH and Ca concentration than the disease conducive soil led us to hypothesize that an increase in soil pH due to the long-term incorporation of Ca-rich organic matter might be the primary cause of the disease suppression. We have designed a highly reproducible bioassay system to examine this hypothesis. The suppressive and conducive soils were mixed with the resting spores of P. brassicae at a rate of 10⁶ spore g⁻¹ soil, and Brassica campestris was grown in a growth chamber for 8 d. The number of root hair infections was assessed on a microscope. It was found that the incorporation of FYM and FSC at 2.5% (w/w) to the conducive soil suppressed the infection and that the finer particles (?5 mm) of FSC inhibited the infection and increased soil pH more effectively. Neutralization of the conducive soil by Ca(OH)₂, CaCO₃ and KOH suppressed the infection, but the effectiveness of KOH was less than those of Ca(OH)₂ and CaCO₃. Acidification of the suppressive soils by H₂SO₄, promoted the infection. The involvement of soil biota in the disease suppression was investigated using the sterilized (γ-ray irradiation) suppressive soils with respect to soil pH. The γ-ray irradiation promoted the infection at pH 5.5, but no infection was observed at pH 7.4 irrespective of the sterilization status. All these observations suggest that soil pH is a major factor in disease suppression by organic matter application and that Ca and soil biota play certain roles in the suppression under the influence of soil pH.     

大白菜根肿病发生规律与防治

根据室内生测及大田试验结果得知,百菌清是控制大白菜根肿病发生的有效药剂。直播大白菜:用60％百菌清可湿性粉剂500g/667m2,与200kg细沙拌匀,毒土法穴施, 之后用60％百菌清可湿性粉剂750倍液(800mg／kg)药液100mL灌根2次,连续3次施药, 间隔期10d,对大白菜根肿病的防效可达70％以上。     

一株多黏类芽胞杆菌 Paenibacillus polymyxa 菌株P1防治广东菜心根肿病的研究

芸薹根肿病是一种危害极大的世界性病害。对此病害的防治多采用化学药剂,但过多地使用化学农药,不仅会导致病原菌的抗药性和药剂防效的不稳定,还会造成环境污染和食品安全等问题。筛选生防菌株并将其开发成有效的生物农药,是防治根肿病并保持绿色农业可持续发展的新途径。本研究选用了多种生防菌对芸薹根肿菌Plasmodiophora brassicae Woron.的休眠孢子进行拮抗测试,筛选出一株能抑制P.brassicae休眠孢子萌发的多黏类芽胞杆菌Paenibacillus polymyxa菌株P1。通过液体发酵和剂型处理,将P1制成悬浮剂(P1-SC),用于室内生测和大田试验,测定其对广东菜心根肿病的防治效果。盆栽试验结果表明,对菜心根肿病的防治效果与所使用的P1-SC浓度呈正相关,当施用P1-SC中P1的浓度≥1×10~6 cfu/mL时,对菜心根肿病的防治效果(83.0%)与化学农药氟啶胺(500 mg/L)的防治效果(100.0%)没有显著性差异,但显著高于(P≤0.05)一种商品枯草芽孢杆菌(2×10~7 cfu/mL)的防治效果(29.8%)。2018年的大田试验结果表明,P1-SC处...     

土壤pH对芸薹根肿菌侵染及病害发生的影响

通过设置不同pH梯度,研究土壤pH对根肿菌侵染及病害发生的影响。结果表明:土壤酸性时病菌侵染速度快,碱性时慢,而强酸性和碱性土壤条件则抑制孢子萌发;pH为6.0时最有利于根肿菌休眠孢子萌发,萌发率最高,为53.96%;碱性条件可使初级原生质团变形凝结成球状,不能正常分裂或延迟形成游动孢子囊,从而不利于根肿菌侵染。白菜发病率与病情指数随pH升高,呈先上升后下降趋势。其中,pH为5.0时,发病率和病情指数最高,pH 7.0~8.0时发病轻。因此,适宜的偏酸性环境条件下,通过作用于病菌休眠孢子萌发和侵染,提高病害危害程度,而中性或碱性条件干扰该过程并降低病害发生。