Diversity of Avr‐vnt1 and AvrSmira1 effector genes in Polish and Norwegian populations of Phytophthora infestans

The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr‐vnt1 effector, recognized by the potato Rpi‐phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi‐Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C‐terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large‐scale cultivation of plants containing the Rpi‐Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi‐phu1 gene were found, the corresponding Avr‐vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.     

Inoculation of Malus genotypes with a set of Erwinia amylovora strains indicates a gene‐for‐gene relationship between the effector gene eop1 and both Malus floribunda 821 and Malus ‘Evereste’

The Gram‐negative bacterium Erwinia amylovora, causal agent of fire blight disease in pome fruit trees, encodes a type three secretion system (T3SS) that translocates effector proteins into plant cells that collectively function to suppress host defences and enable pathogenesis. Until now, there has only been limited knowledge about the interaction of effector proteins and host resistance presented in several wild Malus species. This study tested disease responses in several Malus wild species with a set of effector deletion mutant strains and several highly virulent E. amylovora strains, which are assumed to influence the host resistance response of fire blight‐resistant Malus species. The findings confirm earlier studies that deletion of the T3SS abolished virulence of the pathogen. Furthermore, a new gene‐for‐gene relationship was established between the effector protein Eop1 and the fire blight resistant ornamental apple cultivar Evereste and the wild species Malus floribunda 821. The results presented here provide new insights into the host–pathogen interactions between Malus sp. and E. amylovora.     

Quantification of Mirafiori lettuce big‐vein virus and its vector,Olpidium virulentus,from soil using real‐time PCR

Big vein disease of lettuce (Lactuca sativa) is an economically important disease transmitted through soil by Olpidium virulentus, and has occurred in most production areas worldwide. The disease is assumed to be caused by Mirafiori lettuce big‐vein virus (MiLBVV). To understand the dynamics of the virus and its vector, MiLBVV and O. virulentus were directly detected in soil. DNA and RNA were extracted from 5 g soil using a bead beating method, followed by purification using adsorption to a column. Detection and quantification were performed using real‐time PCR and a TaqMan probe that was prepared based on the CP region of MiLBVV and the rDNA‐ITS region of O. virulentus, respectively. Furthermore, using a visual assessment of the incidence rate of big vein disease on lettuce in agricultural fields, the C_t values of MiLBVV and O. virulentus from soil were also determined using real‐time PCR. The results showed that MiLBVV concentrations in the soil were high in the field, as also determined by a visual assessment of the incidence rate of big vein disease on lettuce. However, the amount of O. virulentus in soil was not directly correlated with the incidence of MiLBVV. From these results, it is suggested that the risk of lettuce crops developing big vein disease can be estimated using an index of the amount of MiLBVV in the soil.     

Using chemical seed dormancy breakers with herbicides for weed management in soyabean and wheat

Variable dormancies result in periodicity in the germination of weeds and make weed control a repetitive practice. Under some conditions, repeated applications of selective herbicides can lead to the dominance of perennial weeds like Cyperus rotundus . Our hypothesis was that applying a chemical dormancy breaker (DB ) plus herbicide mixture would better control a mixture of weed species. Three experiments were designed to develop a cost‐effective DB treatment and to evaluate its dose with herbicides tank‐mixtures for effective weed management. KNO ₃ and gibberellic acid GA ₃ as dormancy breakers offered comparable effects, but KNO ₃ was more economical than GA ₃. KNO ₃ at a 6% concentration was more effective in promoting weed germination than a 3% concentration in soyabean. A combination of KNO ₃ (6%) and pre‐emergence pendimethalin 0.75 kg a.i. ha^?1 + imazethapyr 0.10 kg a.i. ha^?1 controlled annual weeds by 99% and reduced C. rotundus growth by 83%. This treatment gave significantly higher soyabean yield and net returns. Similarly, a tank‐mixture comprising of clodinafop 0.06 kg a.i. ha^?1 + metsulfuron 0.006 kga.i. ha^?1 was more effective against weeds than pre‐emergence tank‐mix application of pendimethalin 0.75 kg a.i. ha^?1 + carfentrazone‐ethyl 0.02 kg a.i. ha^?1 and isoproturon 0.75 kg a.i. ha^?1. The use of pre‐emergence tank‐mixture of pendimethalin 0.75 kg a.i. ha^?1 + imazethapyr 0.10 kg a.i. ha^?1 should exhaust seed/tuber bank if repeated and reduce the application cost of herbicides by 50% and the dose, residue and cost of pendimethalin by 25%.     

Alteration of gene expression profile in the roots of wild diploid Arachis duranensis inoculated with Ralstonia solanacearum

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of peanut (Arachis hypogaea) in China. However, the molecular basis of peanut resistance to R. solanacearum is poorly understood. Arachis duranensis, a wild diploid species of the genus Arachis, has been proven to be resistant to bacterial wilt, and thus holds valuable potential for understanding the mechanism of resistance to bacterial wilt and genetic improvement of peanut disease resistance. Here, suppression subtractive hybridization (SSH) and macroarray hybridization were employed to detect differentially expressed genes (DEGs) in the roots of A. duranensis after R. solanacearum inoculation. A total of 317 unique genes were obtained, 265 of which had homologues and functional annotations. KEGG analysis revealed that a large proportion of these unigenes are mainly involved in the biosynthesis of phytoalexins, particularly in the biosynthetic pathways of terpenoids and flavonoids. Subsequent real‐time polymerase chain reaction (PCR) analysis showed that the terpenoid and flavonoid synthesis‐related genes showed higher expression levels in a resistant genotype of A. duranensis than in a susceptible genotype, indicating that the terpenoids and flavonoids probably played a fundamental role in the resistance of A. duranensis to R. solanacearum. This study provides an overview of the gene expression profile in the roots of wild Arachis species in response to R. solanacearum infection. Moreover, the related candidate genes are also valuable for the further study of the molecular mechanisms of resistance to R. solanacearum.     

Necrotic diseases caused by viruses in Swedish potato tubers

Potato tuber necrosis in the form of spraing symptoms is caused by infection with Potato mop‐top virus (PMTV) or Tobacco rattle virus (TRV); spraing has become more important in the Swedish potato crop production. In this study, the presence in Sweden of three potato‐infecting viruses associated with necrotic symptoms in tubers was demonstrated: PMTV, TRV and Tobacco necrosis virus (TNV). This study shows that both PMTV and TRV are frequent in Swedish potato fields. PMTV was found in all Swedish counties, except the two most northern ones. TRV was present at several locations in southern and up to the central parts of Sweden. Both viruses were found further north than observed in earlier surveys. PMTV and TRV were analysed in tubers with spraing symptoms and it was not possible to decide visually if the symptoms were caused by either PMTV or TRV. Furthermore, a high occurrence of symptomless tuber infections was observed for several potato cultivars. A unique observation from this study was a mixed infection of both PMTV and TRV in tubers of cv. Berber, where no visual differences on symptom development were detectable for these tubers compared to single infections. During the survey, tubers of cv. Melody were found to display necrotic symptoms in the skin that are characteristic for the ABC disease. This suggested infection by TNV, which also could be confirmed.     

Characterization of Dickeya strains isolated from potato grown under hot‐climate conditions

Dickeya strains isolated in Israel in 2006–2010 were characterized by dnaX sequence analysis, pulsed‐field gel electrophoresis (PFGE), biochemical assays and pectolytic activity, and found to be homogeneous: most of them could be classified as ‘Dickeya solani’. Of the 34 strains isolated from imported seed tubers or potato plants grown from imported seed, 32 were typed as ‘D. solani’ and only two were characterized as Dickeya dianthicola. Biovar typing indicated that all ‘D. solani’ strains were biovar 3. ‘Dickeya solani’ strains were most closely related to Dickeya dadantii subsp. dieffenbachiae according to PFGE and dnaX analyses and both species exhibited high pectolytic activity. Expression levels of two putative virulence genes, pelL (encoding a pectic enzyme) and dspE (encoding a type III effector) were significantly induced in ‘D. solani’ strains isolated from potato plants or tubers grown in hot climates such as the Negev region in Israel, compared to those isolated from seed tubers imported from the Netherlands, France or Germany. Results of this study support the hypothesis that ‘D. solani’ strains isolated in Israel are also clonal; however, they appear to be more virulent than strains isolated in Europe.     

Reverse modelling to estimate yield losses caused by crop diseases

Yield loss analysis is critical to inform tactical and strategic decisions in crop health management, and requires quantification of three elements: the levels of injury caused by disease or pest, the actual (injured) yield, and the attainable (uninjured) yield. Reverse modelling allows reconstruction of an object or a process from limited information combined with a mathematical model. This approach is applied to estimate yield losses caused by diseases in winter wheat using a process‐based simulation model (WHEATPEST), in combination with field data generated by a network of experiments across France, where multiple disease injuries and actual yields, but not attainable yields, were measured. The analysis covers 70 [year × region × variety × crop management] combinations encompassing five years (2004–2008), four French regions, two winter wheat varieties (one high‐yielding and one hardy variety), and two levels of crop management corresponding to two levels of chemical intensification. The analysis involved three main successive modelling steps where actual yield, attainable yield and yield losses associated with individual diseases were simulated. Overall, simulated yield losses to combined diseases ranged from 0 to 4.2 t ha^?1, and averaged 0.80 t ha^?1. Septoria tritici blotch caused the highest mean yield loss of 0.66 t ha^?1. The results highlight the contribution of varietal improvement to agricultural sustainability and performances. Reverse modelling can be applied to other crops and diseases or pests, in order to estimate individual and combined disease yield losses.     

Reducing tillage intensity affects the cumulative emergence dynamics of annual grass weeds in winter cereals

Annual grass weeds such as Apera spica‐venti and Vulpia myuros are promoted in non‐inversion tillage systems and winter cereal‐based crop rotations. Unsatisfactory weed control in these conditions is often associated with a poor understanding of the emergence pattern of these weed species. The aim of this study was to investigate, understand and model the cumulative emergence patterns of A. spica‐venti, V. myuros and Poa annua in winter cereals grown in three primary tillage regimes: (i) mouldboard ploughing, (ii) pre‐sowing tine cultivation to 8–10 cm soil depth and (iii) direct drilling. Direct drilling delayed the cumulative emergence of A. spica‐venti and V. myuros (counted together) in contrast with ploughing, while the emergence pattern of P. annua was unaffected by the type of tillage system. The total density of emerged weed seedlings varied between the tillage systems and years with a higher total emergence seen under direct drilling, followed by pre‐sowing tine cultivation and ploughing. The emergence patterns of all species were differently influenced by the tillage systems, suggesting that under direct drilling, in which these species occur simultaneously, management interventions should first and foremost consider that A. spica‐venti and V. myuros emerge over a longer period to avoid control failures.     

Silencing of Erwinia amylovora sy69 AHL‐quorum sensing by a Bacillus simplex AHL‐inducible aiiA gene encoding a zinc‐dependent N‐acyl‐homoserine lactonase

Quorum sensing in Gram‐negative bacteria is regulated by diffusible signal molecules called N‐acyl‐l ‐homoserine lactones (AHLs). These molecules are degraded by lactonases. In this study, six Bacillus simplex isolates were characterized and identified as a new quorum‐quenching species of Bacillus. An aiiA gene encoding an AHL‐lactonase was identified based on evidence that: (i) it showed high homology with other aiiA genes of Bacillus sp.; (ii) the deduced amino acid sequence contained two conserved regions, ¹⁰⁴SHLHFDH¹¹¹ and ¹⁶⁵TPGHTPGH¹⁷³, characteristic of the metallo‐β‐lactamase superfamily; and (iii) the protein had zinc‐dependent AHL‐degrading activity. Additionally, the expression of the aiiA gene was significantly up‐regulated by 3‐oxo‐AHL. The AHL‐lactonase inhibited multiplication of the 3‐oxo‐C6‐AHL‐producing plant pathogen Erwinia amylovora sy69 both in vitro and in planta. The results provide support for the use of the quorum‐quenching functionality of B. simplex in the integrated control of the devastating fire blight pathogen.     

Modelling the effects of herbicide dose and weed density on rice‐weed competition

The effects of herbicide dose on rice‐weed competition were investigated to develop a combined model, which can be utilised to estimate an optimum herbicide dose for a given weed density in paddy rice cultivation. Field studies were conducted in Suwon for rice‐Echinochloa crus‐galli competition and Iksan for rice‐Eleocharis kuroguwai during 2007. The competitive effect of the weeds E. crus‐galli and E. kuroguwai decreased with increasing doses of flucetosulfuron and azimsulfuron, respectively, in the same manner as the standard dose–response curve. The combination of the rectangular hyperbolic model and the standard dose–response curve adequately described the complex effects of herbicide dose and weed competition on rice yield. Parameter estimates were used with the model to predict rice yield and estimate the doses of flucetosulfuron and azimsulfuron required to restrict rice yield loss caused by E. crus‐galli and E. kuroguwai, respectively, to an acceptable level. For a rice yield of 5.0 t ha^?1, the model recommended flucetosulfuron doses of 8.7, 13.4 and 20.1 g a.i. ha^?1 when infested with E. crus‐galli at 12, 24 and 48 plants m^?2 respectively. For a rice yield of 5.2 t ha^?1, the model recommended azimsulfuron doses of 3.9, 7.5 and 12.6 g a.i. ha^?1 when infested with E. kuroguwai at 24, 48 and 96 plants m^?2 respectively. The theoretical outputs of the combined model appear robust and indicate there are opportunities for reduced herbicide use in the field. These now require evaluation under field conditions.     

Barley lesion mimics,supersusceptible or highly resistant to leaf rust and net blotch

Lesion mimic mutants of plants have the feature of spontaneously displaying necrotic spots or bands on their leaves. Lesion mimics have often displayed enhanced resistance to biotrophic pathogens whilst showing increased susceptibility to necrotrophs. This paper identifies three novel, non‐allelic mutants of barley (Hordeum vulgare), which spontaneously form necrotic leaf lesions: Necrotic leaf spot 9.3091 (nec9.3091), Mottled leaf 8.1661 (mtt8.1661) and Mottled leaf 9.2721 (mtt9.2721). The Necrotic leaf spot 8.3550 mutant (nec8.3550), formerly known as bst1, was included in the study because it is a lesion mimic mutant belonging to the same original pool. The reactions of the mutants to the biotroph Puccinia hordei and the necrotroph Pyrenophora teres f. sp. teres were investigated. Mutants nec8.3550 and mtt8.1661 were more resistant than the parental Bowman near‐isogenic line with the Rph3.c gene (Bowman Rph3.c, NGB 22452) to leaf rust, caused by P. hordei. Mutants nec8.3550, mtt8.1661 and mtt9.2721 were more susceptible than Bowman Rph3.c to net blotch, caused by P. teres f. sp. teres. Autofluorescence was detected in leaf tissues of all mutants. Based on the high expression of the PR1 and Hv‐HIR genes, combined with the low susceptibility to P. hordei, nec8.3550 appears to have entered a state of systemic acquired resistance, which is quite distinct from the resistance expressed in mtt8.1661. The latter mutant has low or no expression of PR1 and Hv‐HIR genes, yet it is highly resistant to rust. It is also extremely susceptible to net blotch. These mutants can serve as genetic sources of novel disease resistance for barley improvement.     

Potential of Ypt1 and ITS gene regions for the detection of Phytophthora species in a lab‐on‐a‐chip DNA hybridization array

A novel DNA‐chip hybridization assay that uses the ras‐related GTP‐binding protein 1 gene (Ypt1) was developed for the identification of several devastating Phytophthora species. The hybridization was conducted in a portable microfluidic lab‐on‐a‐chip device for fast and accurate detection of 40 Phytophthora, two Pythium and one Phytopythium species. Moreover, the functionality of the Ypt1 region was examined in comparison to an array for the internal transcribed spacer (ITS) region by in silico modelling. The difference in species‐specific capture probe sequences was lower for the ITS than for the Ypt1 region. While ITS‐probes of Phytophthora ramorum, Phytophthora fragariae and Phytophthora lateralis cross‐reacted with up to 11 non‐target species, Ypt1‐probes were specific except for P. fragariae/Phytophthora rubi. First analyses of artificially inoculated Rhododendron leaves successfully demonstrated the usability of the respective capture probes for the Ypt1 and the ras‐related plant protein Rab1a gene region. The on‐chip hybridization enabled the detection of up to 1 pg μL^?1 target DNA depending on the species examined. Due to the complementarity of ITS and Ypt1 genetic features, the use of multiple loci is recommended to identify targets of different taxonomic rank.     

Fusarium species and chemotypes associated with fusarium head blight and fusarium root rot on wheat in Sardinia

Environmental conditions in Sardinia (Tyrrhenian Islands) are conducive to fusarium root rot (FRR) and fusarium head blight (FHB). A monitoring survey on wheat was carried out from 2001 to 2013, investigating relations among these diseases and their causal agents. FHB was more frequently encountered in the most recent years while FRR was constantly present throughout the monitored period. By assessing the population composition of the causal agents as well as their genetic chemotypes and EF‐1α polymorphisms, the study examined whether the two diseases could be differentially associated to a species or a population. Fusarium culmorum chemotypes caused both diseases and were detected at different abundances (88% 3‐ADON, 12% NIV). Fusarium graminearum (15‐ADON genetic chemotype) appeared only recently (2013) and in few areas as the causal agent of FHB. In F. culmorum, two haplotypes were identified based on an SNP mutation located 34 bp after the first exon of the EF‐1α partial sequence (60% adenine, 40% thymine); the two populations did not segregate with the chemotype but the A‐haplotype was significantly associated with FRR in the Sardinian data set (P = 0·001), suggesting a possible fitness advantage of the A‐haplotype in the establishment of FRR that was neither dependent on the sampling location nor the sampling year. The SNP determining the Sardinian haplotype is distributed worldwide. The question whether the A‐haplotype segregates with characters facilitating FRR establishment will require further validation on a specifically sampled international data set.     

Meta‐analytic modelling of the incidence–yield and incidence–sclerotial production relationships in soybean white mould epidemics

White mould (Sclerotinia sclerotiorum) is a destructive disease of soybean worldwide. However, little is known of its impact on soybean production in Brazil. A meta‐analytic approach was used to assess the relationship between disease incidence and soybean yield (35 trials) and between incidence and sclerotia production (29 trials) in experiments conducted in 14 locations across four seasons. Region, site elevation and season included as moderators in random‐effects and random‐coefficients models did not significantly explain the variability in the slopes of the incidence–yield relationship. The Pearson's r, obtained from back‐transforming the Fisher's Z estimated by an overall random‐effects model, showed that incidence of white mould was moderately and negatively correlated with yield (r = ?0.76, P < 0.0001). A random‐coefficients model estimated a slope of ?17.2 kg ha^?1%^?1, for a mean attainable yield of 3455 kg ha^?1, indicating that a 10% increase in white mould incidence would result in a mean yield reduction of 172 kg ha^?1. White mould incidence and production of sclerotia were strongly and positively correlated (r = 0.85, P < 0.0001). For every 10% increase in white mould incidence, 1 kg ha^?1 of sclerotia was produced. The relationship between disease incidence and production of sclerotia was stronger in southern regions and at higher elevation. In the absence of management, economic losses associated with white mould epidemics, assuming 43% incidence in 22% of the soybean area, were estimated at approximately US $1.47 billion annually within Brazil.     

Cultivar‐dependent partial resistance and associated defence mechanisms in wheat against Zymoseptoria tritici

Septoria tritici blotch caused by the fungus Zymoseptoria tritici is one of the most devastating foliar diseases of wheat. Knowledge regarding mechanisms involved in resistance against this disease is required to breed durable resistances. This study compared the expression of defence and pathogenicity determinants in three cultivars in semicontrolled culture conditions. The most susceptible cultivar, Alixan, presented higher necrosis and pycnidia density levels than Altigo, the most resistant one. In Premio, a moderately resistant cultivar, necrosis developed as in Alixan, while pycnidia developed as in Altigo. In noninfectious conditions, genes coding for PR1 (pr1), glucanase (gluc) and allene oxide synthase (aos) were constitutively expressed at a higher level in both Altigo and Premio than in Alixan, while chitinase2 (chit2), phenylalanine ammonia‐lyase (pal), peroxidase (pox2) and oxalate oxidase (oxo) were expressed at a higher level in Premio only. Except for aos, all genes were induced in Alixan during the first steps of the symptomless infection phase. Only pox2, oxo, gluc and pal genes in Altigo and pal, chs and lox genes in Premio were up‐regulated at some time points. Basal cultivar‐dependent resistance against Z. tritici could therefore be explained by various gene expression patterns rather than high expression levels of given genes. During the necrotrophic phase, Z. tritici cell wall‐degrading enzyme activity levels were lower in Altigo and Premio than in Alixan, and were associated more with pycnidia than with necrosis. Similar tissue colonization occurred in the three cultivars, suggesting an inhibition of the switch to the necrotrophic lifestyle in Altigo.     

Simultaneous monitoring and quantification of Melampsora allii‐populina and Melampsora larici‐populina on infected poplar leaves using a duplex real‐time PCR assay

The rust fungi Melampsora larici‐populina (Mlp) and Melampsora allii‐populina (Map) are the main phytosanitary constraints for commercial poplar cultivation in Europe. Although Mlp is more aggressive and prevalent than Map, the two species may co‐infect the same poplar tree or even the same poplar leaf, making the epidemiological surveys of each species difficult to achieve. In this study, a new duplex real‐time PCR assay targeting each species was developed, based on single‐copy genes. This test proved to be specific, inclusive and was successfully used to detect and quantify each species, starting from urediniospore samples or directly from infected poplar leaves, with or without visible symptoms. This new molecular tool was also assessed for comparative studies of time‐course infection experiments on artificially inoculated poplar leaf discs. These studies showed that the growth dynamics of Map were significantly slower when the two species were co‐inoculated on the same leaf disc, confirming that Map is less aggressive than Mlp.     

Increased resistance to Verticillium dahliae in Arabidopsis plants defective in auxin signalling

Auxin signalling and transport participate in plant–microbe interactions as positive or negative regulators of disease resistance. The present study investigated the responses of Arabidopsis thaliana plants impaired in the auxin receptors TIR1, AFB1 and AFB3 and the auxin transporter AXR4, upon infection by the soilborne root pathogen Verticillium dahliae. Fewer symptoms were recorded in afb1, afb3 and axr4 plants compared to the wild type (wt). qPCR analysis revealed that the decrease in symptom severity in afb1, afb3 and axr4 was correlated with reduction in the growth of the pathogen in the plants. Therefore, afb1, afb3 and axr4 are partially resistant to V. dahliae. Upon V. dahliae infection, the expression of TIR1, AFB1, AFB3 and AXR4 was up‐regulated in roots, while indole‐3‐acetic acid levels were similar to mocks. The partial resistance of afb1, afb3 and axr4 against V. dahliae can be attributed in part to the up‐regulation of defence‐related genes, as it was observed that afb1 and axr4 had higher PR1 levels than wt, while afb3 had higher PDF1.2 levels than wt. The findings of the present study suggest that the auxin signalling defective mutants afb1, afb3 and axr4 show increased resistance against V. dahliae.     

Molecular characterization,comparison of screening methods,and evaluation of cross‐pathogenicity of black rot (Xanthomonas campestris pv. campestris) strains from cabbage,choy sum,leafy mustard and pak choi from Taiwan

Choy sum (Brassica rapa var. parachinensis), leafy mustard (Brassica juncea) and pak choi (B. rapa var. chinensis) are highly nutritious components of diets in Taiwan and other Asian countries, and bacterial black rot caused by Xanthomonas campestris pv. campestris (Xcc) is a major biotic constraint in these crops. As very little was known about the Xcc strains from these crops in these regions, including their cross‐pathogenicity and aggressiveness on different hosts, Xcc strains were obtained from cabbage (Brassica oleracea var. capitata), choy sum, leafy mustard and pak choi crops in Taiwan. Two previously published PCR‐based assays reliably distinguished the Xcc strains from other Xanthomonas species and subspecies. Phylogenetic analysis based on repetitive sequence‐based PCR assays placed the Xcc strains in a clade distinct from other Xanthomonas species, and also showed host specificity. Although all of the Xcc strains from the different host species were pathogenic on all five Brassica test species in both a detached leaf assay and an intact plant assay, in the intact plant assay they showed differences in virulence or aggression on the different test hosts. The Xcc strains from leafy mustard and pak choi were consistently highly aggressive on all the test host genotypes, but the strains from choy sum and cabbage were less aggressive on leafy mustard and choy sum. The intact plant assay proved more discriminating and reliable than the detached leaf assay for comparing the aggressiveness of Xcc strains on different host genotypes, and so, with the new Xcc strains isolated in this study, will be useful for screening leafy brassica germplasm accessions for resistance to black rot.