Pathotypic diversity of Hyaloperonospora brassicae collected from Brassica oleracea

Downy mildew caused by Hyaloperonospora brassicae is an economically destructive disease of brassica crops in many growing regions throughout the world. Specialised pathogenicity of downy mildews from different Brassica species and closely related ornamental or wild relatives has been described from host range studies. Pathotypic variation amongst Hyaloperonospora brassicae isolates from Brassica oleracea has also been described; however, a standard set of B. oleracea lines that could enable reproducible classification of H. brassicae pathotypes was poorly developed. For this purpose, we examined the use of eight genetically refined host lines derived from our previous collaborative work on downy mildew resistance as a differential set to characterise pathotypes in the European population of H. brassicae. Interaction phenotypes for each combination of isolate and host line were assessed following drop inoculation of cotyledons and a spectrum of seven phenotypes was observed based on the level of sporulation on cotyledons and visible host responses. Two host lines were resistant or moderately resistant to the entire collection of isolates, and another was universally susceptible. Five lines showed differential responses to the H. brassicae isolates. A minimum of six pathotypes and five major effect resistance genes are proposed to explain all of the observed interaction phenotypes. The B. oleracea lines from this study can be useful for monitoring pathotype frequencies in H. brassicae populations in the same or other vegetable growing regions, and to assess the potential durability of disease control from different combinations of the predicted downy mildew resistance genes.     

乌鲁木齐地区甘蓝蚜发生规律初报

甘蓝蚜[Brevicoryne brassicae (Linné)]是烏魯木齐地区十字花科蔬菜上最主要的蚜虫。从9月下旬开始,在晚甘藍、晚苤藍及大白菜等蔬菜靠近地面的根、茎凹陷处,以及叶柄基部和叶片上产卵。蚜卵既可随寄主在菜窖里越冬;也可在菜株残体上露地越冬。露地越冬蚜卵,一般不能成为早春的有效蚜源,因其孵化率极低;孵化出来的少数干母,由于不能及时得到食料或因气候不适合,难以存活。越冬蚜卵于4月下旬孵化,5月中旬末产生有翅蚜,主要先在越冬寄生及冬蘿卜留种株上繁殖;到5月下旬至6月初,才陆續迁飞到春、夏十字花科蔬菜及春油菜上,大量繁殖和为害。7月下旬至8月初,晚甘藍与晚苤藍上的蚜虫便成为大白菜和冬蘿卜苗期的主要蚜源。甘藍蚜在烏魯木齐地区,无論是在菜窖里,或是在露地残株上,均不能以成蚜越冬。     

A phylogenetically distinct lineage of Pyrenopeziza brassicae associated with chlorotic leaf spot of Brassicaceae in North America

Light leaf spot, caused by the ascomycete Pyrenopeziza brassicae, is an established disease of Brassicaceae in the United Kingdom (UK), continental Europe, and Oceania (OC, including New Zealand and Australia). The disease was reported in North America (NA) for the first time in 2014 on Brassica spp. in the Willamette Valley of western Oregon, followed by detection in Brassica juncea cover crops and on Brassica rapa weeds in northwestern Washington in 2016. Preliminary DNA sequence data and field observations suggest that isolates of the pathogen present in NA might be distinct from those in the UK, continental Europe, and OC. Comparisons of isolates from these regions using genetic (multilocus sequence analysis, MAT gene sequences, and rep-PCR DNA fingerprinting), pathogenic (B. rapa inoculation studies), biological (sexual compatibility), and morphological (colony and conidial morphology) analyses demonstrated two genetically distinct evolutionary lineages. Lineage 1 comprised isolates from the UK, continental Europe, and OC, and included the P. brassicae type specimen. Lineage 2 contained the NA isolates associated with recent disease outbreaks in the Pacific Northwest region of the USA. Symptoms caused by isolates of the two lineages on B. rapa and B. juncea differed, and therefore “chlorotic leaf spot” is proposed for the disease caused by Lineage 2 isolates of P. brassicae. Isolates of the two lineages differed in genetic diversity as well as sensitivity to the fungicides carbendazim and prothioconazole.     

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.     

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.     

Orobanche palaestina: a potential threat to agricultural crops in Israel

Weeds of the genus Orobanche are obligatory chlorophyll-lacking root parasites that infect and severely damage many dicotyledonous agricultural crops in warm-temperate and subtropical regions of the world. The genus comprises over 100 species, at least six of which are notable weeds. Orobanche palaestina Reut. is an endemic Mediterranean species, previously reported to parasitize thistles [Notobasis syriaca (L.) Cass. and Cirsium phyllocephalum Boiss. & Bl.] and annual legumes in native Mediterranean habitats, mainly in disturbed places such as roadsides and waste sites. In recent years, we have observed a rapid expansion of O. palaestina populations adjacent to commercial agricultural fields in the Jezreel Valley of Israel. The ability of O. palaestina to infect crops was tested in pots under nethouse conditions. The experiments revealed that O. palaestina is able to parasitize agricultural crops. Safflower (Carthamus tinctorius L.), lettuce (Lactuca sativa L.), gazania (Gazania uniflora Gaertn.), vetch (Vicia sativa L.) and artichoke (Cynara scolymus L.) were found to be potential hosts for this parasite, and all of them, except vetch and artichoke, were preferred to thistle, the native host. Sunflower (Helianthus annuus L.) induced O. palaestina seed germination and root attachment, but the attached parasites could not reach maturity and died soon after the attachment stage. Tomato (Lycopersicon esculentum Mill.), carrot (Daucus carota L.), garland chrysanthemum (Chrysanthemum coronarium L.) and cabbage (Brassica oleraceae L.) were not parasitized by O. palaestina.     

Discrimination of <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">campestris</Emphasis> races among strains from northwestern Spain by <Emphasis Type="Italic">Brassica</Emphasis> spp. genotypes and rep-PCR

Black rot, caused by Xanthomonas campestris pv. campestris (Xcc) is a severe seedborne disease of Brassica crops around the world. Nine races are recognized, being races 1 and 4 the most aggressive and widespread. The identification of Xcc races affecting Brassica crops in a target area is necessary to establish adequate control measures and breeding strategies. The objectives of this study were to isolate and identify Xcc strains from northwestern Spain by using semi-selective medium and pathogenicity tests, determine the existing races of Xcc in this area by differential series of Brassica spp., and evaluate the use of repetitive DNA polymerase chain reaction-based fingerprinting (rep-PCR) to differentiate among the nine existing Xcc races. Seventy five isolates recovered from infected fields were identified as Xcc. Race-typing tests determined the presence of the following seven pathogen races: 1, 4, 5, 6, 7, 8 and 9. Race 4 was the most frequent in Brassica oleracea and race 6 in Brassica rapa crops, therefore breeding should be focussed in obtaining resistant varieties to both races. Cluster analysis derived from the combined fingerprints showed four groups, but no clear relationship to race, crop or geographical origin was found. Rep-PCR analysis was found not to be a reliable method to discriminate among Xcc races, therefore race typing of Xcc isolates should be done by using the differential series of Brassica spp. genotypes or another alternative approach.     

Molecular analysis and virus transmission tests place Olpidium virulentus, a vector of Mirafiori lettuce big-vein virus and tobacco stunt virus, as a distinct species rather than a strain of Olpidium brassicae

The rDNA-ITS sequences of ten single-sporangium isolates of Olpidium virulentus (a noncrucifer strain of Olpidium brassicae), which transmits Mirafiori lettuce big-vein virus (MLBVV) and tobacco stunt virus (TStV), were compared with those of six single-sporangium isolates of O. brassicae. The sequence similarity within isolates of O. virulentus or O. brassicae was almost identical (98.5%–100.0%), but was low between the two species (79.7%–81.8%). In a phylogenetic analysis of the rDNA-ITS region, O. virulentus and O. brassicae fell into two distinct clusters, indicating that O. virulentus, a vector of MLBVV and TStV, is a distinct species rather than a strain of O. brassicae.     

Occurrence of Groundnut ringspot virus on Brazilian peanut crops

A high incidence of plants with mosaic, chlorotic spots, ringspots, necrosis, smaller leaves, and stunting was observed on peanut crops (Arachis hypogaea L.) in Itápolis, São Paulo State, Brazil. Transmission electron microscope examination of thin sections of infected leaves revealed the presence of spheroidal particles, ca. 80 nm in diameter, suggestive of Tospovirus. A DNA fragment of ~600 bp was amplified by RT-PCR from total RNA extracted from infected tissues using primers specific for the nucleocapsid gene of Groundnut ringspot virus (GRSV). Nucleotide and deduced amino acid sequences of the fragments showed high identities with known GRSV isolates.     

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.     

In vitro inhibition of mycelial growth of Phytophthora nicotianae Breda de Haan from different hosts by Brassicaceae species. Effect of the developmental stage of the biofumigant plants

BACKGROUND: The hydrolysis products of glucosinolates in Brassicaceae tissues are potentially useful for the control of fungal pathogens. The in vitro activity of these products against Phytophthora nicotianae Breda de Haan was studied, with the isolates exposed to the volatile products released from the brassica tissues. RESULTS: The four species of Brassicaceae tested inhibited the mycelial growth of P. nicotianae isolates. The most effective developmental stage was different, depending on the species: buds yellowing in Sinapis alba L. and Brassica carinata A. Braun; seeds enlarging and all buds open in Brassica nigra (L.) W.D.J. Koch; cauliflower still covered by leaves in Brassica oleracea L. var. botrytis. At this stage, B. nigra and S. alba were the most effective (53.6 and 52.5% inhibition respectively). With all the biofumigants species tested, isolates from pepper plants were more susceptible to the brassica effect than those from tomato isolates. CONCLUSION: The developmental stage of plants has an influence on the biofumigant potential of Brassicaceae species against P. nicotianae. The isolates differ in susceptibility to compounds released, depending on their host of origin, suggesting the differentiation of populations of P. nicotianae in relation to the host of origin. Copyright © 2012 Society of Chemical Industry     

Mapping of clubroot (Plasmodiophora brassicae) resistance in canola (Brassica napus)

Clubroot disease, caused by Plasmodiophora brassicae, has become a major problem in the production of cruciferous crops worldwide. In this study, a population of 121 doubled haploid (DH) lines derived from a crossing between a resistant and a susceptible canola (Brassica napus) genotype was subjected to phenotypic and genotypic studies to determine the inheritance and location of the resistance gene(s). After inoculation with pathotype 3 of P. brassicae, the lines showed a 1:1 segregation ratio for resistance, indicating that resistance in this population is controlled by a single gene. Fifteen PCR‐based markers that were known to be linked to clubroot resistance (CR) genes were screened against genomic DNA from parents and resistant and susceptible bulks. Marker GC1680, linked to the CR gene CRa, exhibited polymorphism between the parents and between the resistant and susceptible bulks. CRa target primers were used to amplify fragments from the two parents and the resultant sequences were compared. A high degree of sequence similarity was found between the parents in the nucleotide binding site domain of CRa. In contrast, sequence polymorphisms were detected in the leucine‐rich repeat (LRR) domain. One pair of primers that amplify a band from the LRR region of the resistant parent but not the susceptible parent was used to screen the DH population. Amplicons were obtained from 60 of the 61 resistant lines and two of the 60 susceptible lines; thus, three recombinants were found. Based on these results, a resistance locus linked to CRa was found.     

Competence of <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> from Cruciferous Weeds and Wallflower (<Emphasis Type="Italic">Erysimum cheiri</Emphasis>) to Induce Black Rot in Cabbage

Aphanomyces euteiches Drechsler is an oomycete pathogen of leguminous crops that causes root rot, a severe disease of pea (Pisum sativum L.) worldwide. An improved understanding of the genetic structure of A. euteiches populations would increase knowledge of pathogen evolution and assist in the design of strategies to develop pea cultivars and germplasm with stable disease resistance. Twenty six primers pairs were used to amplify Sequence Related Amplified Polymorphisms (SRAP) among 49 A. euteiches isolates sampled from pea. A total of 190 polymorphic SRAP bands were generated, of which 82 were polymorphic between all the A. euteiches isolates. The percentage of polymorphic bands per primer pair ranged from 22 to 75%. According to the PIC value estimated for each marker, 60% of the SRAP markers were highly to reasonably informative (PIC > 0.25). Genetic structure of A. euteiches populations sampled in different American and French locations showed low to high genetic diversity within populations. The largest variation occurred within countries, with a total estimated genetic diversity of 0.477 and 0.172 for American and French populations, respectively. This was particularly evident from a principal component analysis (PCA) and a Minimum Spanning Networks (MSN) based on genetic profiles of isolates, which generated two different clusters, one corresponding to the French isolates and four American isolates (MV1, MV5, MV7, Ath3), and the other to American isolates. A. euteiches populations from cultivated pea in France appeared as a single unstructured population, whereas American isolates of A. euteiches diverged into three different populations.     

Inhibition of chitin synthesis by benzoyl-phenylurea insecticides,III. Similarity in action in Pieris brassicae (L.) with Polyoxin D

The increase in cuticle thickness with age of fifth instar larvae of Pieris brassicae (L.) was measured microscopically. The injection of a lethal dose of either Polyoxin D or diflubenzuron revealed total inhibition of cuticular growth and caused comparable abnormalities in the cuticles.In a further experiment [¹⁴C]glucose was injected along with Polyoxin D into Pieris brassicae and the incorporation of radioactivity into various tissue fractions was measured. This revealed that the impairment of cuticular growth was due to inhibition of chitin synthesis.With the methods used the effects of Polyoxin D and two benzoylphenylurea insecticides appeared to be the same.     

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.     

Analysis of genetic and virulence variability of Stemphylium lycopersici associated with leaf spot of vegetable crops

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46?S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.), pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage.     

Development of a Pathotype Specific SCAR Marker in Plasmodiophora brassicae

Plasmodiophora brassicae is an obligate biotroph that causes clubroot, one of the most damaging diseases of crucifers. Breeding of clubroot-resistant plants has been hampered by the large variation of pathogenicity in P. brassicae and by the lack of an efficient means for detecting specific isolates. To improve the practicality of P. brassicae pathotype-identification, a molecular approach was developed. RAPD profiles of 37 single-spore-derived isolates belonging to seven different pathotypes were compared. A RAPD marker, OPL14₁₂₀₀, was found in the molecular pattern of all the isolates belonging to one particular pathotype (P1), pathogenic on all differential hosts tested. The DNA band corresponding to this marker was cloned and sequenced. No significant homology to previously characterised nucleotide sequences was found. Primers were designed to specifically amplify the OPL14₁₂₀₀ band. The SCAR marker was observed in all isolates belonging to pathotype P1 and was absent in isolates belonging to other pathotypes and in the different plant hosts analysed. The SCAR marker was also generated from direct amplification of DNA from clubs (mixture of host and pathogen DNA) developed after infection by P1 isolates. This molecular marker may be a valuable tool for rapid and reliable identification of P. brassicae P1 isolates in areas where resistant varieties are cultivated.     

Epidemiology of root rot caused by <Emphasis Type="Italic">Leptosphaeria maculans</Emphasis> in <Emphasis Type="Italic">Brassica napus</Emphasis> crops

The infection of above-ground tissues of Brassica napus by Leptosphaeria maculans is well understood. However, root infection (root rot) under field conditions, the development of root rot over time and its relationship to other disease symptoms caused by L. maculans has not been described. A survey of B. napus crops was conducted in Australia to investigate the incidence and severity of root rot. Additionally, the pathway of root infection was examined in field experiments. Root rot was present in 95% of the 127 crops surveyed. The severity and incidence of root rot was significantly correlated with that of crown canker; however, the strength of this relationship was dependent on the season. Root rot symptoms appeared before flowering and increased in severity during flowering and at maturity, a pattern similar to crown canker suggesting that the infection of the root is an extension of the crown canker phase of the L. maculans lifecycle. All isolates of L. maculans tested in glasshouse experiments caused root rot and crown canker in B. napus and Brassica juncea. In the field, the main pathway of root infection is via invasion of cotyledons or leaves by airborne ascospores, rather than from inoculum in the soil. Root rot was present in crops in fields that had never been sown to B. napus previously, in plants grown in fumigated fields, and in glasshouse-grown plants inoculated in the hypocotyl with L. maculans.