湖南白菜和红菜薹根肿病菌生理小种鉴定

为了弄清湖南省白菜和红菜薹根肿病菌生理小种组成和分布,2013年采集长沙、湘潭、益阳、怀化4个地区的十字花科作物白菜、红菜薹根肿病株27个,用Williams根肿病鉴别品种对其进行生理小种鉴定。初步鉴定出5个生理小种,即1号、5号、8号、9号和11号,它们在湖南病区中出现的频率分别为22.3%、18.5%、14.8%、33.3%和11.1%。按寄主生理小种组成,湖南白菜根肿病菌生理小种有1号、5号、9号、11号,红菜薹根肿病菌为8号、9号和11号;按地区生理小种分布,长沙白菜根肿病菌生理小种为5号和9号,湘潭白菜和红菜薹根肿病菌生理小种分别1号、8号和11号,怀化白菜根肿病菌为生理小种9号;益阳白菜和红菜薹根肿病菌分别为生理小种1号、11号、8号和9号。目前,在湖南4个市白菜和红菜薹根肿病菌生理小种是9号。     

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。     

云南省近年稻瘟病菌生理小种的组成和分布

用日本 9个水稻单基因鉴别品种和BL1、K59两个参考品种鉴定了云南省 4个稻作区21个县 (市 )采集、分离的 155个稻瘟病菌单孢菌株 ,结果出现 78个稻瘟病菌生理小种 ,其中优势小种为136.4(出现频率为65%)、317.4(出现频率52% )、007(出现频率 5.2% )小种。通过研究各稻作区生理小种对日本鉴别品种的侵染率 ,分析了各垂直抗性基因在不同稻作区的利用价值。     

甘蓝类蔬菜品种(系)苗期根肿病抗性鉴定

芸薹根肿菌侵染引起的根肿病是影响甘蓝类蔬菜产量和品质的主要病害之一,抗病品种的培育是防治根肿病最经济有效的方法。本研究以根肿菌4号生理小种为菌源,对花椰菜、结球甘蓝、青花菜、芥蓝和苤蓝6个甘蓝变种共306份品种(系)进行苗期根肿病抗性鉴定,结果显示,甘蓝类抗根肿病材料缺乏,仅在青花菜中发现1份高抗(HR)材料,感病(S)和高感(HS)材料比例占65%以上,中抗(MR)材料在松花菜和结球甘蓝中所占比例均在30%以上。对抗病比例和平均病情指数2个指标的联合评价结果显示,结球甘蓝整体抗性水平高于其他变种,芥蓝和苤蓝抗性最差。另外,对结球甘蓝‘先甘336’(R)×甘蓝/青花菜杂交后代及花椰菜‘托尼’(MR)×青花菜的杂交后代进行苗期根肿病抗性鉴定,结果表明,杂交后代相对于父本病情指数都有所降低,抗性相对提高,且父本病情指数越低,后代表现较高抗性的概率越大。鉴于此,在今后抗病组合的选择与选配过程中,父本和母本最好都具有较高的抗性。本研究将为甘蓝类蔬菜抗根肿病品种培育提供材料支撑和技术参考。     

广东稻瘟病菌生理小种辅助鉴别寄主的筛选及应用

近年来研究发现,利用中国稻瘟病菌生理小种7个鉴别寄主划分的同一小种不同菌株间的致病性差异较大,这对如何合理筛选代表菌株应用于新品种(组合)的抗性鉴定造成一定困难。作者从广东目前生产上推广面积较大的品种或主要抗源中筛选出4个能较好区分菌株致病性差异的品种(抗源)作为稻瘟病菌生理小种鉴定的辅助鉴别寄主。 1 材料与方法每年在广东不同生态稻区的主栽品种上收集穗颈瘟标样,分离、培养和鉴定方法均按全国统一方法进行。供试水稻品种或资源为广东省的主栽品种以及各育种部门提供的新     

青花菜根肿病苗期抗性鉴定技术的建立

为建立快速、有效的青花菜根肿病苗期抗性鉴定技术,将芸薹根肿菌Plamodiophora brassicae Woron.人工接种于高感根肿病青花菜自交系90196,研究了接种菌液浓度、接种寄主苗龄、接种基质p H和接种方法等对人工接种鉴定效果的影响。结果表明,在一定范围内,根肿病发病率及病情指数随着接种菌液浓度的升高而增大,接种菌液浓度为3×108CFU/m L时,发病率及病情指数分别为97.22%和86.11,可以反映寄主真实的抗性水平;接种寄主苗龄为2~6叶期均能使植株发病,但2~3叶期发病效果最佳;接种基质偏酸性(p H 5~6)有利于根肿病的发生;使用伤根灌菌法进行鉴定,青花菜根肿病发病率和病情指数均最高,分别为100.00%和88.10,优于蘸根法和浸芽法。用已知抗性水平的12个自交系和8个杂交种进行验证,鉴定结果表明该苗期抗性鉴定技术可客观反映供试材料的实际抗性水平。     

我国部分水稻品种对稻瘟病的抗谱测定

选用10省6群16个稻瘟病菌生理小种,对10省29个研究所提供的318个水稻品种(系)进行测定,提出加农Si-Pi661020、81005等19个抗性品种(系)可作杂交亲本利用,对其中经济性状较好的品种(系)可根据各省当地小种的出现频率和分布情况,在较大范围内种植。 本试验选用我国近年分离的16个稻瘟菌生理小种,对318个有希望的水稻品种(系)进行了苗期抗稻瘟病谱测定和田间人工接种、自然诱发3病(稻瘟病、白叶枯病、纹枯病)的鉴定,为全面评价供测品种(系)的抗病性提供依据^[2]。现将试验的方法和结果简述如下:     

江苏省水稻稻瘟病菌生理小种分布及品种抗性测定

江苏省水稻稻瘟病菌生理小种分布及品种抗性测定刁春友，施阿宝（江苏省稻瘟病生理小种监测协作组，２１００３７）近年来，随着我省水稻品种结构的调整，粳稻面积扩大，水稻稻瘟病有明显加重的趋势。省植保站与省农科院植保所及射阳等７个县（市）植保站联合组成江苏省稻...     

糯高粱种质资源对川南高粱丝黑穗病的抗性鉴定

为明确新选育和引进的糯高粱种质资源对川南高粱丝黑穗病菌主要生理小种的抗性,利用4个高粱鉴别寄主分别对2013年和2014年自川南泸州、宜宾、自贡3市高粱上采集的丝黑穗病菌Sporisorium reilianum进行生理小种鉴定,并于2014—2018年对新选育和引进的55份不育系、88份恢复系和20份常规品种糯高粱种植资源进行丝黑穗病抗性鉴定。结果表明,川南3市采集的高粱丝黑穗病菌生理小种均为1号生理小种。不育系糯高粱种植资源对丝黑穗病菌1号生理小种抗性较强,对丝黑穗病免疫材料有45份,87.3%不育系糯高粱种植资源为抗性材料。恢复系糯高粱种质资源抗丝黑穗病菌1号小种的能力弱于不育系,对丝黑穗病表现为免疫、高抗、抗病、中抗的种质资源分别为32、1、2和5份,抗病恢复系糯高粱种质资源占恢复系糯高粱种质资源总数的45.5%。糯高粱种质资源常规品种对丝黑穗病菌1号小种的抗性最差,抗病糯高粱种质资源常规品种占常规品种总数的45.0%,其中对丝黑穗病免疫的糯高粱种质资源常规品种仅有1份,为国窖红3号;对丝黑穗病高抗和抗病的糯高粱种质资源常规品种无,主要表现为中抗,应加强川南地区糯高粱常规品种的丝黑穗病抗性改良。鉴定的对丝黑穗病免疫的糯高粱不育系、恢复系和常规品种可作为川南高粱丝黑穗病病区主栽品种或抗丝黑穗病菌1号生理小种的育种材料。     

中国小麦生产品种对条锈菌不同生理小种抗病性分析

小麦条锈病是严重威胁我国小麦安全生产的病害,抗病品种培育和利用是经济、安全、高效的防控策略.通过对来自全国不同麦区的115个小麦生产品种进行苗期和成株期抗条锈病分小种鉴定的结果显示,供试小麦品种对条锈菌流行小种的抗病性存在明显差异.其中,苗期对7个条锈菌生理小种均表现抗病的品种9个(占参试品种的7.8%),均表现感病的品种23个(占20.0%);对条锈菌生理小种CYR32、CYR33和V26均表现全生育期抗性的品种13个(占11.3%)、成株抗性品种5个(占4.3%)和慢锈性品种3个(占2.6%).表明当前我国小麦主产区品种整体抗条锈性水平仍较低,需大力加强小麦抗条锈病育种工作,并对不同麦区小麦品种合理布局问题进行了讨论.     

Variation in pathotypes and virulence of Plasmodiophora brassicae populations in Germany

Between 2012 and 2015, 49 new clubroot‐infested fields were identified in 12 German federal states. Clubroot disease incidence varied within these fields from 22% to 92%. Field information revealed that in 85% of fields, oilseed rape was grown in rotation once every 2 or 3 years. Frequency of OSR in the rotation was significantly correlated with the incidence and prevalence of clubroot disease. The disease was detected in fields with soil pH ranging from 5.1 to 8.3, and a significant negative correlation was found between soil pH and the disease incidence of infested fields. Furthermore, more cases of disease and severe incidences were observed in sandy loam and loamy sand as compared with other soil types. Pathotype classification of the 49 Plasmodiophora brassicae populations was conducted on two differential sets, the European Clubroot Differential set and the set of Somé (1996). Additionally, the degree of virulence of the collected isolates was analysed on the clubroot‐resistant oilseed rape cv. Mendel. The results showed variation in pathotype distribution in different regions in Germany. The majority of isolates according to Somé were pathotypes 1 and 3, respectively, with pathotypes 2 and 5 in the minority. Detailed classification according to Buczacki showed the dominance of 16/31/31, 16/14/30 and 16/14/31 populations among 20 distinct virulence patterns of collected isolates. From all populations tested for virulence on cv. Mendel, 15 isolates were found to be moderately or highly virulent. These virulent populations were not restricted to a small geographical area in the country.     

Pathogenic and genetic diversity in Plasmodiophora brassicae (clubroot) from Japan

Clubroot disease, caused by Plasmodiophora brassicae Woronin, affects various cruciferous crops. Variations in pathogenicity and virulence are present among field populations of P. brassicae. Many races (pathotypes) have been reported in Japan as well as in other countries using various differential systems. Populations can be classified into four pathotypes using two clubroot-resistant (CR) cultivars of Chinese cabbage as differential hosts in Japan. However, it was recently indicated that each population is often heterogenic and composed of multiple genotypes (races or pathotypes). Breakdown in CR cultivars of Chinese cabbage is a problem in some areas of Japan and may contribute to the selective propagation of minor pathogenic genotypes on the CR cultivars. Clubroot has also been recorded on five species of cruciferous weeds in Japan. In particular, clubroot of Cardamine flexuosa is widely distributed in Japan. Some populations of C. flexuosa are often moderately pathogenic on Chinese cabbage and turnip. Therefore, the epidemiological relationship between clubroot of cruciferous crops and that of the weed has been noted but not thoroughly clarified. The relationship between pathogenic and genetic variations has also been examined among populations from cruciferous crops and weeds in Japan. The result implies an interesting genetic relationship among Williams’ races, among pathotypes determined using CR cultivars of Chinese cabbage and among populations from crops and C. flexuosa. This review includes an introduction of the status of studies on pathogenic and genetic diversity in P. brassicae from Japan.     

Potential loss of clubroot resistance genes from donor parent Brassica rapa subsp. rapifera (ECD 04) during doubled haploid production

Clubroot resistance derived from the oilseed rape/canola Brassica napus ‘Mendel’ has been overcome in some fields in Alberta, Canada, by the emergence of ‘new’ strains of the protist Plasmodiophora brassicae. Resistance to the pathogen was assessed in 112 doubled haploid (DH) lines, derived from B. rapa subsp. rapifera (European clubroot differential (ECD) 04). The lines were evaluated against five single‐spore isolates representing the ‘old’ pathotypes 2, 3, 5, 6 and 8, and 15 field populations representing new strains of P. brassicae. The disease severity index (ID%) data revealed that none of the DH lines were resistant or moderately resistant to the new pathotype 5X (field populations L‐G1, L‐G2, L‐G3) and D‐G3, while 3–42% were resistant or moderately resistant to the other 11 new strains. Using the mean ID induced by the old pathotype 3 (approx. 13.5%) as the baseline, clubroot severity increased by 300–600% when inoculated with the new pathotypes. A significant finding of this study was the fact that ECD 04 showed absolute resistance to all of the old and new P. brassicae strains while the B. napus ‘Mendel’, although resistant to all of the old pathotypes, was resistant to only about 50% of the new strains. Similarly, all of the selected clubroot‐resistant commercial canola cultivars evaluated in this study were susceptible to 87% of the new P. brassicae strains. The molecular data revealed that the breakdown of clubroot resistance in Mendel and the canola cultivars was in part due to the non‐inheritance of the Crr1 gene on the A08 chromosome from ECD 04.     

Virulence and inoculum density‐dependent interactions between clubroot resistant canola (Brassica napus) and Plasmodiophora brassicae

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.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Effect of host and non‐host crops on Plasmodiophora brassicae resting spore concentrations and clubroot of canola

Plasmodiophora brassicae, causal agent of clubroot of crucifers, poses a serious threat to Canadian canola production. The effects of fallow (F) periods and bait crops (clubroot‐susceptible canola (B) and perennial ryegrass (R)) on clubroot severity and P. brassicae resting spore populations were evaluated in five sequences: R–B, B–R, R–F, B–F and F–F. Both host and non‐host bait crops reduced clubroot severity in a subsequent crop of a susceptible canola cultivar compared with fallow. Resting spore and P. brassicae DNA concentrations decreased in all treatments, but were lowest for the R–B and B–R bait crop sequences. In addition, two studies were conducted in mini‐plots under field conditions to assess the effect of rotation of susceptible or resistant canola cultivars on clubroot severity and P. brassicae resting spore populations. One study included three crops of susceptible canola compared with a 2‐year break of oat–pea, barley–pea, wheat–wheat or fallow–fallow. The other study assessed three crops of resistant canola, two crops of resistant canola with a 1‐year break, one crop of resistant canola and a 2‐year break, and a 3‐year break with barley followed by a susceptible canola. The rotations that included non‐host crops of barley, pea or oat reduced clubroot severity and resting spore concentrations, and increased yield, compared with continuous cropping of either resistant or susceptible canola. Growing of a susceptible canola cultivar contributed 23–250‐fold greater gall mass compared with resistant cultivars.     

Plasmodiophora brassicae resting spore dynamics in clubroot resistant canola (Brassica napus) cropping systems

The soilborne pathogen Plasmodiophora brassicae, causal agent of clubroot of canola (Brassica napus), is difficult to manage due to the longevity of its resting spores, ability to produce large amounts of inoculum, and the lack of effective fungicides. The cropping of clubroot resistant (CR) canola cultivars is one of the few effective strategies for clubroot management. This study evaluated the impact of the cultivation of CR canola on P. brassicae resting spore concentrations in commercial cropping systems in Alberta, Canada. Soil was sampled pre-seeding and post-harvest at multiple georeferenced locations within 17 P. brassicae-infested fields over periods of up to 4 years in length. Resting spore concentrations were measured by quantitative PCR analysis, with a subset of samples also evaluated in greenhouse bioassays with a susceptible host. The cultivation of CR canola in soil with quantifiable levels of P. brassicae DNA resulted in increased inoculum loads. There was a notable lag in the release of inoculum after harvest, and quantifiable P. brassicae inoculum peaked in the year following cultivation of CR canola. Rotations that included a ≥2-year break from P. brassicae hosts resulted in significant declines in soil resting spore concentrations. A strong positive relationship was found between the bioassays and qPCR-based estimates of soil infestation. Results suggest that CR canola should not be used to reduce soil inoculum loads, and crop rotations in P. brassicae infested fields should include breaks of at least 2 years away from B. napus, otherwise the risk of selecting for virulent pathotypes may increase.     

Identification of Brassica accessions resistant to ‘old’ and ‘new’ pathotypes of Plasmodiophora brassicae from Canada

Genetic resistance is the main tool used to manage clubroot of canola (Brassica napus) in Canada. However, the emergence of new virulent strains of the clubroot pathogen, Plasmodiophora brassicae, has complicated canola breeding efforts. In this study, 386 Brassica accessions were screened against five single-spore isolates (represented by pathotypes 2F, 3H, 5I, 6M and 8N on the Canadian Clubroot Differential Set) and 17 field isolates (represented by 12 unique pathotypes: 2B, 3A, 3D, 3O, 5C, 5G, 5K, 5L, 5X, 8E, 8J and 8P) of P. brassicae to identify resistance sources effective against these strains. The results showed that one B. rapa accession (CDCNFG-046, mean index of disease (ID) = 3.3%) and two B. nigra accessions (CDCNFG-263, mean ID = 3.1%; and CDCNFG-262, mean ID = 4.7%) possessed excellent resistance to all 22 of the isolates evaluated. Fifty other accessions showed differential clubroot reactions (resistant, moderately resistant or susceptible), including 27 (one B. napus, two B. rapa, four B. oleracea and 20 B. nigra) accessions that were each resistant to 8–21 P. brassicae isolates, but developed mean IDs in the range of 5.3–29.6%. The remaining 23 accessions (two B. napus, one B. rapa, five B. oleracea and 15 B. nigra) were each resistant to 3–13 isolates, but developed mean IDs in the range of 30.3–47.0%. The three accessions that showed absolute resistance and the 50 accessions that showed differential clubroot reactions could be used to breed for resistance to the new P. brassicae strains.     

Resistance to infection by stealth: <Emphasis Type="Italic">Brassica napus</Emphasis> (winter oilseed rape) and <Emphasis Type="Italic">Pyrenopeziza brassicae</Emphasis> (light leaf spot)

Light leaf spot (Pyrenopeziza brassicae) is an important disease on winter oilseed rape crops (Brassica napus) in northern Europe. In regions where economically damaging epidemics occur, resistance to P. brassicae in commercial cultivars is generally insufficient to control the disease without the use of fungicides. Two major genes for resistance have been identified in seedling experiments, which may operate by decreasing colonisation of B. napus leaf tissues and P. brassicae sporulation. Much of the resistance present in current commercial cultivars is thought to be minor gene-mediated and, in crops, disease escape and tolerance also operate. The subtle strategy of the pathogen means that early colonisation of host tissues is asymptomatic, so a range of techniques and molecular tools is required to investigate mechanisms of resistance. Whilst resistance of new cultivars needs to be assessed in field experiments where they are exposed to populations of P. brassicae under natural conditions, such experiments provide little insight into components of resistance. Genetic components are best assessed in controlled environment experiments with single spore (genetically fixed) P. brassicae isolates. Data for cultivars used in the UK Recommended List trials over several seasons demonstrate how the efficacy of cultivar resistance can be reduced when they are deployed on a widespread scale. There is a need to improve understanding of the components of resistance to P. brassicae to guide the development of breeding and deployment strategies for sustainable management of resistance to P. brassicae in Europe.     

Development of simple sequence repeat markers for the classification of the clubroot pathogen <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

Clubroot disease caused by Plasmodiophora brassicae is one of the most serious diseases in cruciferous crops. To classify isolates, we developed simple sequence repeat (SSR) markers for P. brassicae. Twenty-four Japanese isolates were used in this study: 12 isolates of an unknown pathotype from the Kyoto Prefecture, as well as 12 isolates of known pathotypes, including three single-spore lines. From the 12 isolates from Kyoto Prefecture, 11 were classified into either pathotype 2 (three isolates) or 4 (eight isolates). We designed 23 SSR markers based on the P. brassicae genome, of which 11 markers from intergenic regions showed polymorphisms in the 24 isolates. Many haploid isolates belonging to pathotypes 2 and 4 were monomorphic, and typical alleles were detected in some isolates not belonging to pathotype 4. Two bands were detected for eight SSR loci in five isolates, indicating that different genotypes were mixed in these isolates. We constructed a phylogram based on the 11 polymorphic SSRs. Pathotypes 2 and 4 formed a cluster, from which pathotypes 3 and 1 were successively placed. These results strongly suggest a close genetic relationship between isolates in pathotypes 2 and 4, consistent with our finding that isolates in these two pathotypes were found at one collection site. In combination with pathotype classification and other marker systems, the SSR markers can be used for more detailed analyses to improve the control of clubroot disease.