Sequence analyses of Wheat dwarf virus isolates from different hosts reveal low genetic diversity within the wheat strain

Wheat dwarf virus (WDV) causes disease in wheat (Triticum aestivum) and barley (Hordeum vulgare) in many parts of Europe. The host range also includes many species of the family Poaceae. WDV is only transmitted by the leafhopper Psammotettix alienus. During a five‐year period (2001–2005), grass samples were collected in central Sweden in the vicinity of fields with WDV‐infected winter wheat. Screening with ELISA and PCR identified WDV in a low number of samples (8/1098) from only three grass species: Apera spica‐venti, Avena fatua and Poa pratensis. In addition, triticale was found to be positive. Fourteen WDV isolates from Avena fatua, Apera spica‐venti, Triticum aestivum, Lolium multiflorum, Poa pratensis, triticale and the insect vector Psammotettix alienus, were partially sequenced (ca. 1200 nucleotides), providing the first published WDV sequences from the insect vector. All isolates belonged to the wheat strain of WDV and the genetic diversity was low. Phylogenetic analyses showed no clear grouping according to geographical location or host species. The results suggest that the same WDV genotypes are infecting both wheat and grasses in Sweden. Interestingly, one group of isolates (subtype B) formed a distinct clade in the phylogenetic tree. Subtype B was always found in mixed infection with the main genotype. Complete sequencing of a subtype B isolate showed that it was 98·6% identical to a typical wheat isolate from the same plant.     

Slow wilting components in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis>(L.) Millsp.)

Partial resistance to Fusarium wilt was characterized based on root inoculation on seven pigeonpea genotypes with a virulent isolate of Fusarium udum. Resistance to wilt seems to be mainly due to localization of the pathogen in roots and vascular systems in the stem. Three components of resistance (wilting rate or incubation period, weighted wilt index and number of colonized plants) were determined and compared to those of susceptible genotypes Bahar and TTB7. There were significant differences between the genotypes for the resistance components. The mechanisms of resistance in the genotypes appeared to be different, with genotype ICP8863, having a longer incubation period, minimum wilt index and minimum pathogen colonization as compared to other resistant genotypes (ICP9174, ICP87119 and ICP8858). Wilting rate or incubation period and number of colonized plant were significantly correlated with resistance in adult plants in the field (AUDPC). Wilt index was useful in discriminating between genotypes that had a similar incubation period and number of colonized plants. The partial character of resistance is probably based on quantitative differences in localization capacity among the genotypes. A quantitative relationship between components, incubation period and number of colonized plants and the AUDPC, if verified for a large number of genotypes, may be used to obtain an index of resistance that may predict resistance levels in the field.     

Effects of host and pathogen genotypes on inducibility of resistance in tomato (Solanum lycopersicum) to Phytophthora infestans

Thirteen tomato (Solanum lycopersicum) accessions were tested for inducibility of resistance against two isolates of Phytophthora infestans using BABA (dl ‐3‐amino butyric acid) as the inducing agent. In a more detailed trial, six of the accessions were assessed for inducibility of resistance to six P. infestans isolates on three leaves of different age per plant. Plants were inoculated 1 week after treatment with BABA. Area under the disease progress curve (AUDPC), sporulation capacity (SC) and infection efficiency (IE) were all affected by treatment with BABA. On leaves of all ages AUDPC was most affected by induction (43–100% reduction on the youngest leaves) followed by SC (14–100%) and IE (0–100% reduction). Tomato genotypes varied significantly in inducibility of resistance against P. infestans and the degree of induction generally decreased with increasing leaf age, whilst the absolute susceptibility with respect to AUDPC and SC rarely changed. The level of induction was not always related to the resistance level of the tomato accession and it was significantly influenced by the pathogen isolate used for challenge inoculation. The results show that inducibility of resistance is a selectable trait that is, however, isolate‐specific.     

Identification of isolate‐specific resistance QTLs to phytophthora root rot using an intraspecific recombinant inbred line population of pepper (Capsicum annuum)

Quantitative trait loci (QTL) for resistance to phytophthora root rot caused by Phytophthora capsici were investigated using two Korean P. capsici isolates and 126 F₈ recombinant inbred lines derived from a cross of Capsicum annuum line YCM334 (resistant parent) and local cv. Tean (susceptible parent). The experimental design was a split plot with two replications. Highly significant effects of pathogen isolate, plant genotype, and genotype × isolate were detected. QTL mapping was performed using a genetic linkage map covering 1486·6 cM of the pepper genome, and consisted of 249 markers including 136 AFLPs (Amplified Fragment Length Polymorphisms), 112 SSRs (Simple Sequence Repeats) and one CAPS (Cleaved Amplified Polymorphic Sequence). Fifteen QTLs were detected on chromosomes 5 (P5), 10 (P10), 11 (P11), Pb and Pc using two data processing methods: percentage of wilted plants (PWP) and relative area under the disease progress curves (RAUDPC). The phenotypic variation explained by each QTL (R²) ranged from 6·0% to 48·2%. Seven QTLs were common to resistance for the two isolates on chromosome 5 (P5); six were isolate‐specific for isolate 09‐051 on chromosomes 10 (P10) and Pc, and two for isolate 07‐127 on chromosomes 11 (P11) and Pb. The QTLs in common with the major effect on the resistance for two isolates explained 20·0–48·2% of phenotypic variation. The isolate‐specific QTLs explained 6·0–17·4% of phenotypic variation. The result confirms a gene‐for‐gene relationship between C. annuum and P. capsici for root rot resistance.     

Phytophthora infestans populations in central,eastern and southern African countries consist of two major clonal lineages

Limited knowledge is available on Phytophthora infestans populations in Sub‐Saharan Africa (SSA). Therefore, and in response to recent severe late blight epidemics, P. infestans isolates from potato, tomato and Petunia × hybrida from eight SSA countries were characterized. Isolates were characterized with ‘old’ markers, including mating type (176 isolates), mitochondrial DNA haplotype (mtDNA) (281 isolates), glucose‐6‐phosphate isomerase (Gpi) (70 isolates), restriction fragment length polymorphism analysis with probe RG‐57 (49 isolates), and by metalaxyl sensitivity (64 isolates). Most isolates belonged to the US‐1 genotype or its variants (US‐1.10 and US‐1.11). The exceptions were genotype KE‐1 isolates (A1 mating type, mtDNA haplotype Ia, Gpi 90/100 and unique RG‐57 genotype), identified in two fields in Kenya, which are related to genotypes previously identified in Rwanda (RW‐1 and RW‐2), Ecuador and Europe. Metalaxyl‐resistant P. infestans isolates from potato were present in all the countries except Malawi, whereas all the isolates from tomato were sensitive. Genotyping of 176 isolates with seven simple sequence repeat (SSR) markers, including locus D13 that was difficult to score, revealed 79 multilocus genotypes (MLGs) in SSA. When this locus was excluded, 35 MLGs were identified. Genetic differentiation estimates between regional populations from SAA were significant when locus D13 was either excluded (P = 0·05) or included (P = 0·007), but population differentiation was only low to moderate (F_ST = 0·044 and 0·053, respectively).     

Pathogenic variation of Mycosphaerella species infecting banana and plantain in Nigeria

Mycosphaerella species that cause the ‘Sigatoka disease complex’ account for significant yield losses in banana and plantain worldwide. Disease surveys were conducted in the humid forest (HF) and derived savanna (DS) agroecological zones from 2004 to 2006 to determine the distribution of the disease and variation among Mycosphaerella species in Nigeria. Disease prevalence and severity were higher in the HF than in the DS zone, but significant (P < 0·001) differences between agroecological zones were only observed for disease severity. A total of 85 isolates of M. fijiensis and 11 isolates of M. eumusae were collected during the survey and used to characterize the pathogenic structure of Mycosphaerella spp. using a putative host differential cultivar set consisting of Calcutta‐4 (resistant), Valery (intermediate) and Agbagba (highly susceptible). Area under disease progress curve (AUDPC) was higher on all cultivars when inoculated with M. eumusae than with M. fijiensis, but significant (P < 0·05) differences between the two species were only observed on Valery. Based on the rank‐sum method, 8·3% of the isolates were classified as highly aggressive and 46·9% were classified as aggressive. About 11·5% of all the isolates were classified as least aggressive, and all of these were M. fijiensis. The majority of M. eumusae isolates (seven out of 11; 64%) were classified as aggressive. A total of nine pathotype clusters were identified using cluster analysis of AUDPC. At least one M. fijiensis isolate was present in all the nine pathotype clusters, while isolates of M. eumusae were present in six of the nine clusters. Isolates in pathotype clusters III and V were the most aggressive, while those in cluster VIII were the least aggressive. Shannon’s index (H) revealed a more diverse Mycosphaerella collection in the DS zone (H = 1·81) than in the HF (H = 1·50) zone, with M. fijiensis being more diverse than M. eumusae. These results describe the current pathotype structure of Mycosphaerella in Nigeria and provide a useful resource that will facilitate screening of newly developed Musa genotypes for resistance against two important leaf spot diseases of banana and plantain.     

Characterization of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melongenae</Emphasis> isolates from eggplant in Turkey by pathogenicity,VCG and RAPD analysis

Fusarium wilt is an economically important fungal disease of common eggplant (Solanum melongena) cultivated in the eastern Mediterranean region of Turkey. Seventy-four isolates of Fusarium oxysporum isolated from diseased eggplant displaying typical Fusarium wilt symptoms were screened for pathogenicity on the highly susceptible cv. ‘Pala’. All the isolates tested were pathogenic to eggplant and designated as Fusarium oxysporum f. sp. melongenae (Fomg). Genetic diversity among a core set of 20 Fomg isolates that were selected based upon geographic locations, were characterized by using pathogenicity, vegetative compatibility grouping (VCG), and random amplified polymorphic DNA (RAPD) analysis. The area under the disease progress curve (AUDPC) was calculated for each Fomg isolate until 21 days after inoculation (DAI). The most virulent isolate was identified as Fomg10 based on AUDPC, disease severity and vascular discoloration measurements at 21 DAI. At this date, a good correlation was observed between disease severity and AUDPC values for all isolates (r = 0.73). UPGMA (unweighted pair group method with arithmetic average) cluster analysis of RAPD data using Dice’s coefficient of similarity differentiated all the Fomg isolates tested, and indicated considerable genetic variation among Fomg isolates, but isolates from the same geographic region were grouped together. There was no direct correlation between clustering in the RAPD dendrogram and pathogenicity testing of Fomg isolates. Twenty isolates of Fomg were assigned to VCG 0320.     

Quantitative trait loci associated with resistance to a potato isolate of Verticillium albo‐atrum in Medicago truncatula

Verticillium albo‐atrum is responsible for considerable yield losses in many economically important crops, among them alfalfa (Medicago sativa). Using Medicago truncatula as a model for studying resistance and susceptibility to V. albo‐atrum, previous work has identified genetic variability and major resistance quantitative trait loci (QTLs) to Verticillium. In order to study the genetic control of resistance to a non‐legume isolate of this pathogen, a population of recombinant inbred lines (RILs) from a cross between resistant line F83005.5 and susceptible line A17 was inoculated with a potato isolate of V. albo‐atrum, LPP0323. High genetic variability and transgressive segregation for resistance to LPP0323 were observed among RILs. Heritabilites were found to be 0·63 for area under the disease progress curve (AUDPC) and 0·93 for maximum symptom score (MSS). A set of four QTLs associated with resistance towards LPP0323 was detected for the parameters MSS and AUDPC. The phenotypic variance explained by each QTL (R²) was moderate, ranging from 4 to 21%. Additive gene effects showed that favourable alleles for resistance all came from the resistant parent. The four QTLs are distinct from those described for an alfalfa V. albo‐atrum isolate, confirming the existence of several resistance mechanisms in this species. None of the QTLs co‐localized with regions involved in resistance against other pathogens in M. truncatula.     

Species composition,toxigenic potential and pathogenicity of Fusarium graminearum species complex isolates from southern Brazilian rice

This study aimed to assess the extent and distribution of Fusarium graminearum species complex (FGSC) diversity in rice seeds produced in southern Brazil. Four species and two trichothecene genotypes were detected among 89 FGSC isolates, based on a multilocus genotyping assay: F. asiaticum (69·6%) with the nivalenol (NIV) genotype, F. graminearum (14·6%) with the 15‐acetyldeoxynivalenol (ADON) genotype, and F. cortaderiae (14·6%) and F. meridionale (1·1%), both with the NIV genotype. Seven selected F. asiaticum isolates from rice produced NIV in rice‐based substrate in vitro, at levels ranging from 4·7 to 84·1 μg g^?1. Similarly, two F. graminearum isolates from rice produced mainly 15‐ADON (c. 15–41 μg g^?1) and a smaller amount of 3‐ADON (c. 6–12 μg g^?1). One F. meridionale and two F. cortaderiae isolates did not produce detectable levels of trichothecenes. Two F. asiaticum isolates from rice and two from wheat (from a previous study), and one F. graminearum isolate from wheat, were pathogenic to both crops at various levels of aggressiveness based on measures of disease severity in wheat spikes and rice kernel infection in a greenhouse assay. Fusarium asiaticum and the reference F. graminearum isolate from wheat produced NIV, and deoxynivalenol and acetylates, respectively, in the kernels of inoculated wheat heads. No trichothecene was produced in kernels from inoculated rice panicles by any of the isolates. These findings constitute the first report of FGSC composition in rice outside Asia, and confirm the dominance of F. asiaticum in rice agroecosystems.     

Fusarium solani f. sp. passiflorae: a new forma specialis causing collar rot in yellow passion fruit

The aim of this study was to characterize a Fusarium population obtained from yellow passion fruit (YPF) with collar rot using pathogenicity, morphocultural characteristics and molecular tests. Pathogenicity and disease severity were assessed in six plant species: YPF, zucchini, tomato, bean, soya bean and cucumber. Potato dextrose agar medium (PDA) was used to determine mycelial growth at five temperatures (15–35°C). The colour produced by isolates was also determined on PDA at 25°C. Synthetic nutrient agar medium was used to evaluate: (i) type of mycelium and phialides; (ii) size, shape and number of septa from conidia; and (iii) production of chlamydospores and perithecia. Molecular tests consisted of sequencing the ITS–5·8S rDNA region and elongation factor 1α (EF‐1α) gene. The isolates caused large lesions on YPF, zucchini and tomato, with YPF having the highest mean disease severity and being the only one that showed wilt symptoms and death of the plant. Thus the isolates showed host specificity. Maximum mycelial growth occurred at 25°C and the predominant colour was bluish‐white. The isolates produced long phialides, dense aerial mycelium, oval microconidia with a mean size of 9·5 × 2·6 μm, macroconidia of 32·7 × 3·4 μm with 3·3 septa, and chlamydospores; only one isolate lacked perithecia. Phylogenetic trees of the ITS region and EF‐1α gene showed that isolates from YPF formed a distinct group within the F. solani group and the formae speciales of F. solani. It is proposed to name all isolates from YPF as F. solani f. sp. passiflorae.     

Evaluation of different methods for the characterization of carrot resistance to the alternaria leaf blight pathogen (Alternaria dauci) revealed two qualitatively different resistances

Alternaria leaf blight (ALB), caused by Alternaria dauci, is one of the most damaging foliar diseases of carrot worldwide. The aim of this study was to compare different methods for evaluating levels of carrot resistance to ALB. Three techniques were investigated by comparison with a visual disease assessment control: in vivo conidial germination, a bioassay based on a drop‐inoculation method, and in planta quantification of fungal biomass by quantitative PCR (Q‐PCR). Three carrot cultivars showing different degrees of resistance to A. dauci were used, i.e. a susceptible cultivar (Presto) and two partially resistant genotypes (Texto and Bolero), challenged with an aggressive or a very aggressive isolate of A. dauci. Both partially resistant genotypes produced a higher mean number of germ tubes per conidium (up to 3·42±0·35) than the susceptible one (1·26±0·18). The drop‐inoculation results allowed one of the partially resistant genotypes (Bolero, log₁₀(S+1) = 1·34±0·13) to be distinguished from the susceptible one (1·90±0·13). By contrast, fungal growth measured by Q‐PCR clearly differentiated the two partially resistant genotypes with log₁₀(I) values of 2·77±0·13 compared to the susceptible cultivar (3·65±0·13) at 15 days post‐inoculation. This result was strongly correlated (r² = 0·91) with the disease severity index scored at the same date. Data obtained with the different assessment methods strongly suggest that the Texto and Bolero genotypes have different genetic resistance sources.     

Relationship between the extent of colonization by Verticillium dahliae and symptom expression in strawberry (Fragaria × ananassa) genotypes resistant to verticillium wilt

Eleven strawberry (Fragaria × ananassa) genotypes from the University of California breeding programme known to be resistant to verticillium wilt were inoculated with Verticillium dahliae. Individual plants were given a resistance score based on the severity of visual symptoms, and the extent of colonization was quantified as the percentage of petioles not colonized by the pathogen. Both resistance scores and the percentage of pathogen‐free petioles decreased significantly from May to June (P < 0·05) during each of two growing seasons, indicating a progression of both colonization and symptom expression. Even the most resistant genotypes had plants with some infected petioles, and manifested some symptoms of verticillium wilt. Significant (P < 0·05) genotypic variance was detected for the percentage of pathogen‐free petioles, but not for resistance score. The percentage of pathogen‐free petioles had a strongly positive genotypic correlation (r_g = 0·77, P < 0·01) with resistance score, indicating that about 60% of the genotypic variation for visual symptoms in this set of resistant genotypes was explained by the extent of colonization of individual plants by V. dahliae. Conversely, the genotypic correlation between the percentage of pathogen‐free petioles and the resistance score for plants sampled in May (r_g = 0·74, P < 0·01) was smaller than that for plants harvested in July (r_g = 0·93, P < 0·01). Together, these results suggest that the overall performance of strawberry genotypes in the presence of V. dahliae can be enhanced by both resistance and tolerance, but that tolerance may be less stable over the course of a season. Distinguishing between these two mechanisms may require evaluations that supplement visual assessments of resistance.     

Russian populations of Puccinia triticina in distant regions are not differentiated for virulence and molecular genotype

The objective of this study was to determine whether genetically distinct groups of Puccinia triticina are present in four regions of the Russian Federation. Collections of P. triticina were obtained from the Central, North Caucasus, Volga and West Siberia regions from 2006 to 2010. Ninety‐nine single uredinial isolates were tested for virulence phenotype with 20 Thatcher near‐isogenic lines of wheat. Forty‐one virulence phenotypes were found in the four regions, with eight in common between the widely separated Central and West Siberia regions. A total of 72 isolates were tested for molecular genotype with 23 simple sequence repeat (SSR) primer pairs, and 66 isolates were used for further analysis after clone correction for virulence and molecular genotype. Analysis of variation showed no overall differentiation of SSR genotypes or virulence phenotypes based on region of origin. Linkage disequilibria for SSR genotypes were high across the entire population. The regional populations had higher than expected levels of allelic heterozygosity that indicated clonal reproduction. Based on cluster analysis of SSR genotypes there were two groups of P. triticina isolates that were widely distributed across Russia. The two SSR groups also differed significantly for virulence. Puccinia triticina may be dispersed from a common source of inoculum in the European or Caucasus regions of Russia. The Russian P. triticina populations were highly differentiated for SSR genotype from populations in Tajikistan, Kyrgyzstan, Uzbekistan, Armenia, Georgia and Azerbaijan and more similar to populations from southern Kazakhstan and northern Kazakhstan.     

Assessment of resistance to <Emphasis Type="Italic">Ceratocystis cacaofunesta</Emphasis> in cacao genotypes

Ceratocystis wilt of cacao (caused by Ceratocystis cacaofunesta) is a dangerous disease and results in the death of the plant. This fungus was recently identified in the major cacao-producing regions of Brazil, and was observed to be more aggressive than isolates from other geographical locations. The objective of this study was to develop and test a consistent method to assess cacao genotype response to C. cacaofunesta, based on young plants (seedlings or cuttings). The fungus was inoculated by the deposition of propagule suspensions on cut stems. The parameters to assess disease progress were (a) disease incidence, (b) differences in mortality between the most contrasting cacao genotypes for resistance and susceptibility, (c) disease index, (d) consistency of response over time and (e) relative lesion heights. When seedlings were used for the analyses, the ICS-1 and TSH-1188 genotypes proved to be useful as genetic standards for susceptibility and resistance to C. cacaofunesta, respectively. Inoculum concentrations between 10⁴ and 10⁵ propagules ml⁻¹ and the moment at which the disease incidence stabilized provided appropriate conditions for genotypic comparison. When ten cacao genotypes propagated by cuttings (clones) were assessed, the results confirmed TSH-1188 as the reference genotype for resistance to C. cacaofunesta, while the remaining clones could be grouped as resistant (CEPEC-2008), moderately resistant (CEPEC-2002, CEPEC-2007) and susceptible (CEPEC-2009, CCN-10, CCN-51, HW-25, PH-16, SJ-02). The analytical concepts and results were discussed in terms of their application in breeding programmes aimed at developing genetic resistance to Ceratocystis wilt of cacao.     

Interaction between silicon amendment,bacterial wilt development and phenotype of Ralstonia solanacearum in tomato genotypes

Silicon amendment significantly reduced bacterial wilt incidence expressed as area under disease progress curve for tomato genotypes L390 (susceptible) by 26.8% and King Kong2 (moderately resistant) by 56.1% compared to non-treated plants grown in hydroponic culture. However, wilt incidence in silicon-treated plants of genotype L390 reached 100% at 13 days post-inoculation (dpi), while in genotype King Kong2, plant death was retarded by 6 days, with 20% reduction of final wilt incidence. Bacterial numbers were significantly lower in silicon-treated compared to non-treated plants in King Kong2 at 2 dpi in midstems and in all organs at 5 dpi, and in Hawaii 7998 (resistant) in all organs at 2 dpi. Differences between genotypes were obvious on midstem level (5 dpi), where bacterial populations were generally significantly lower compared to roots. Increased tolerance was observed in genotypes L390 and King Kong2 with silicon treatment.Silicon accumulated in roots and was low in stems and leaves. Inoculation with Ralstonia solanacearum did not significantly affect silicon uptake and distribution. Negative correlations between root silicon content and bacterial numbers of midstems in genotypes Hawaii 7998 and King Kong2 suggested an induced resistance. Indications for an influence of host genotype and silicon treatment on the phenotypic conversion of R. solanacearum strain To-udk2-sb from fluidal to non-fluidal colonies in planta were observed.This is the first report on the effect of silicon on a bacterial disease and in a silicon-non-accumulator plant.     

High level resistance to Pseudocercosporella capsellae offers new opportunities to deploy host resistance to effectively manage white leaf spot disease across major cruciferous crops

Field and controlled environment studies were undertaken to define the range and extent of available host resistances to Pseudocercosporella capsellae (white leaf spot) across diverse oilseed, forage and vegetable crucifers, including some wild and/or weedy species, and also within and/or derived from Brassica carinata. In each experiment, there was a wide range in host response from high resistance to high susceptibility as assessed by four disease parameters, viz. in the field for: (i) Area Under Disease Progress Curve (AUDPC) for percent leaves diseased with values ranging from 0 to 375.5; (ii) Percent Leaf Collapse Index (%LCI) for leaf collapse due to disease with values ranging from 0 to 23.0; and (iii), Percent Pod Area Disease Index (%PADI) for pod area affected with values ranging from 0 to 52.1; and (iv) under controlled environmental conditions for Percent Cotyledon Disease Index (%CDI) for cotyledon lesion size with values ranging from 0 to 27.5. At the Crawley field site, B. carinata ATC 94129 was the most resistant genotype with AUDPC?=?1.2, followed by Crambe abyssinica (AUDPC 8.7), Eruca sativa Eruc-01 (AUDPC 19.3) and E. vesicaria Yellow rocket (AUDPC 19.4). B. carinata ATC 94129 and B. oleracea var. capitata had the least leaf collapse, with %LCI?=?0.2. At the Shenton Park field site, 21 genotypes of B. carinata and B. oleracea var. acephala Tuscan kale showed total resistance, all with AUDPC values of 0. Of the B. napus genotypes carrying one or more B. carinata B genome introgressions, genotypes NC8 (AUDPC 23.0) and NC9-1 (AUDPC 26.2) were the most resistant. Genotypes as assessed on these disease criteria as having high level resistance generally showed no pod infection; in contrast to %PADI values up to 52 on the most susceptible genotypes. Under controlled environmental conditions, the most resistant genotype was B. carinata ATC 94129 with %CDI values of 0 and 0.2, respectively, across two experiments, along with B. napus genotypes Zhongyou 821 and Hyola 42, with a %CDI value of 0 in one of the two experiments. There was a high degree of correlation both within individual experiments across the different disease parameters and also between field and controlled environment experiments. Within both B. napus and B. juncea genotypes tested, the most resistant genotypes were from China, the most susceptible from India, with those from Australia intermediate.     

Response of strawberry genotypes to inoculation with isolates of Verticillium dahliae differing in host origin

Eight strawberry genotypes known to differ in susceptibility to verticillium wilt were inoculated with eight isolates of Verticillium dahliae originally obtained from six different host crops: strawberry, potato, watermelon, mint, eggplant (aubergine) and cauliflower. Inoculation experiments were conducted in replicated field trials during two successive years. Known susceptible genotypes developed typical symptoms of verticillium wilt in both years. Although isolates manifested a wide range of aggressiveness, differences were significant only on the most susceptible strawberry genotype. Two isolates originating from strawberry were among the most aggressive of the eight tested, whereas the least aggressive isolate was obtained from cauliflower. Six strawberry genotypes that were regarded as resistant to verticillium wilt based on previous tests were also resistant in the present study, regardless of the isolate used. Overall, strawberry genotypes represented the largest source of variation in these experiments, with variance components approximately 10-fold greater than those associated with either isolate or the isolate × genotype interaction. The results suggest it should be possible to develop resistance to verticillium wilt in strawberry that is broadly effective against isolates of diverse host origin.     

High genotype diversity and lack of isolation by distance in the Alternaria solani populations from China

A genomic library was used to develop seven SSR markers for studying the population genetics of Alternaria solani, a pathogenic fungus causing early blight disease of potato and tomato worldwide. Population genetic analysis of 268 isolates of A. solani sampled from four locations, each representing one of four potato production systems in China, indicates that these SSR markers are moderately diverse, selectively neutral and possibly unlinked. Population genetic analysis also indicated that genetic variation of A. solani in China is high. About 2/3 of 123 genotypes were detected only once and genotype diversity measured by the standardized Shannon index ranged between 0·82 and 0·92 in the populations. Although clones were detected in multiple populations separated by thousands of kilometres, random association among SSR loci was found in half of the populations assayed. On average, nearly six copies of genetic material were exchanged among these populations each generation and no isolation by distance was detected. It is hypothesized that the joint effects of cryptic sexual reproduction and human‐mediated gene flow may account for the observed population genetic structure of A. solani in China.     

Genotyping Cephalosporium gramineum and development of a marker for molecular diagnosis

This study investigated the genetic variation of 40 isolates of Cephalosporium gramineum, the causal agent of cephalosporium stripe disease of wheat, based on variations in internal transcribed spacers (ITS) and intergenic spacers (IGS) of rDNA. Of the isolates, 29 were from Japan and the rest from the USA and Europe. The ITS region was about 600 bp and almost identical among these isolates. In the IGS region (～5 kbp), restriction fragment length polymorphism analysis detected four genotypes among the 40 isolates. One representative isolate was selected from each of the four genotypes, and the IGS region was sequenced. Attempts to design a genotype‐specific marker based on the size of PCR products amplified with selected primers failed to differentiate among the four genotypes. Alternatively, a species‐specific primer set (CGIGS1 and CGIGS2) was developed that annealed within the conserved region, producing a DNA fragment of about 1·8 kbp. Tests of this primer set on a wide range of other fungi from 11 genera confirmed that it was specific to C. gramineum. This primer set could serve as an effective tool in the molecular diagnosis of C. gramineum and has the potential to assist in a better understanding of the host–pathogen interaction.