Genotype-isolate interaction for resistance to black stem in sunflower (Helianthus annuus)

Two experiments were undertaken to determine the partial resistance of sunflower genotypes to seven isolates of Phoma macdonaldii . In the first experiment, 28 genotypes, including recombinant inbred lines and their parents, M6 mutant lines developed by gamma irradiation, and some genotypes from different geographical origins, were used. The experiment consisted of a split-plot design with three replications, each with 12 seedlings per genotype per isolate, in controlled conditions. Seven days after inoculation, plantlets were scored on a 1–9 scale for percentage necrotic area. Highly significant differences were observed among genotypes, isolates and their interactions. The presence of a differential interaction between genotypes and P. macdonaldii isolates was confirmed in a second experiment using 12 genotypes representing large variability for partial resistance to P. macdonaldii isolates, as identified in the first experiment. Inbred lines B454/03, ENSAT-B5 and LC1064C were the most susceptible sunflower genotypes, whereas two American lines SDR19 and SDR18 presented high partial resistance to all P. macdonaldii isolates studied. The least and most aggressive isolates were MA6 and MP3, respectively. Isolates interacted differentially with sunflower genotypes. This study identified two genotypes (AS613 and PAC2) presenting specific resistance to isolate MP8. The results also showed a wide range of isolate-nonspecific partial resistances among the lines tested. The information presented here could assist sunflower breeders to choose parents of crosses for breeding of durable resistance to phoma black stem disease.     

水稻与稻瘟病菌非亲和性互作中重要防御酶活性变化规律研究

 选以CO39为背景的水稻抗稻瘟病近等基因系,与稻瘟菌生理小种ZC₁₃(菌株97-151a)组成的3类典型非亲和性互作,以亲和性互作为对照,对各互作中过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶及β-1,3-葡聚糖酶的活性变化规律进行了系统研究。完全非亲和性互作C101A51/97-151a、高度非亲和性互作C101L AC/97-151a及中度非亲和性互作C104 PKT/97-151a,POD比活性接种后即开始明显升高,48h前达到高峰,升高趋势一直持续到7d完全显症时,幅度基本与各互作非亲和程度呈正相关;亲和性互作CO39/97-151a接种后40 h POD比活性才开始升高,4~6 d达到高峰,峰值也较大。3类非亲和性互作PAL比活性在接种后0 h或16 h开始较明显升高,整个互作中形成3~4个较明显的峰;亲和性互作中PAL比活性一直明显下降。3类非亲和性互作外切几丁质酶比活性接种后即开始升高,基本一直保持升高趋势,在40 h前幅度较大,并形成1~3个较高的峰;亲和性互作外切几丁质酶比活性接种后即开始大幅度升高直至完全显症,48h后幅度远高于非亲和性互作。3类非亲和性互作β-1,3-葡聚糖酶比活性在24 h内开始较明显升高,在48h前形成2~3个较明显的峰;亲和性互作在接种后β-1,3-葡聚糖酶比活性即开始升高,在48h后显著高于非亲和性互作。讨论了POD、PAL、几丁质酶及β-1,3-葡聚糖酶参与水稻抗稻瘟病的可能性。     

Molecular analysis of resistance mechanisms to Orobanche cumana in sunflower

Resistance to the dicotyledenous parasite Orobanche cumana in sunflower is characterized by a low number of parasitic attachments and a confinement of the parasite in host tissues leading to its necrosis. To help understand what determines such resistance mechanisms, molecular, biochemical and histological approaches were employed before (early response) and after (late response) attachment of the broomrape parasite to susceptible (2603) and resistant (LR1) sunflower genotypes. The expression patterns of 11 defence-related genes known to be involved in different metabolic pathways (phenylpropanoids, jasmonate, ethylene) and/or in resistance mechanisms against microorganisms were investigated. RT-PCR and cDNA blot experiments revealed that the resistant genotype exhibited a stronger overall defence response against O. cumana than the susceptible one, involving marker genes of the jasmonate (JA) and salicylic acid (SA) pathways. Among them, the SA-responsive gene, def. (defensin), appeared to be characteristic of LR1 sunflower resistance. However, no JA accumulation and similar SA contents (250–300 ng g⁻¹ FW) were measured by GC/MS in both genotypes, parasitized or not. In addition, three cDNAs, isolated by a suppression-subtractive hybridization, were shown to be strongly induced only in the resistant genotype 8 days post-inoculation, when the first O. cumana attachments occurred. These genes, putatively encoding a methionine synthase, a glutathione S-transferase and a quinone oxidoreductase, might be involved in detoxification of reactive oxygen species, suggesting the occurrence of an oxidative burst during the incompatible interaction. Finally, host cell-wall modifications leading to parasite-confinement were correlated with more intense callose depositions in the resistant genotype, concomitant with over-expression of the callose synthase cDNA HaGSL1 .     

Microtubule dynamics in compatible and incompatible interactions of soybean hypocotyl cells with Phytophthora sojae

The arrangement of microtubules in soybean ( Glycine max ) cells was examined during compatible and incompatible interactions of hypocotyls of soybean cv. Harosoy (susceptible) and cv. Haro 1272 (resistant) with race 1 of the soybean-specific pathogen Phytophthora sojae . Both reaction types were similar during the first 3 h after zoospore inoculation in terms of the number of cells penetrated, and depth penetrated into the cortex. By 3 h postinoculation, clear differences had developed between the two interaction types: incompatible interactions were characterized by a hypersensitive response that was confined to single penetrated cells; while compatibly responding cells appeared unchanged. Both types of response were characterized by autofluorescence of cell walls or cytoplasm and, at 6 h after inoculation, complete disorganization of cell cytoplasm. Reorientation and loss of microtubules was seen in the early stages of the incompatible interaction in association with cellular hypersensitivity, but not in compatible responses. In cells adjacent to those that reacted hypersensitively, there was little evidence of change in microtubule orientation. Treatment of hypocotyls with the microtubule depolymerizer oryzalin prior to inoculation did not alter the compatible response, but led to breakdown of the incompatible response. Changes in microtubule orientation and state are thus among the first structural changes that are visible within cells during incompatibility in this system.     

Characterisation of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> isolates collected from potato in Estonia during 2002–2003

A collection of 101 isolates of Phytophthora infestans, obtained from seven sampling sites representing central, east and south-east Estonia during 2002 and 2003 were assessed for several phenotypic and genotypic markers. All 101 isolates were assessed for virulence and resistance to metalaxyl. Virulence to each of the 11 classic resistance genes was found among the tested isolates. The mean number of virulences per isolate was 6.3, with a very low frequency of virulence against resistance genes R5 (5%) and R9 (14%). The most common pathotypes were 1.3.4.7.8.10.11 and 1.3.4.7.10.11, representing altogether 12% of the studied strains. In terms of metalaxyl resistance, 30 resistant, 52 intermediate and 19 sensitive isolates were found. A subgroup of 50 isolates was assessed for mating type, allozymes [glucose-6-phosphate isomerase (Gpi) and peptidase (Pep)], DNA fingerprints with probe RG57 and mtDNA haplotype. Of this subset, 30 were A1 and 20 were A2. Collections from three of the seven fields contained both mating types. Allozyme analysis did not reveal any polymorphism. However, 19 diverse RG57 fingerprints were detected, and two mitochondrial DNA haplotypes, Ia and IIa, were detected. By combining the mating type, mtDNA haplotype and RG57 fingerprint data, 26 multilocus genotypes were identified, of which 18 were detected only once. Genotypic diversity measured by the normalised Shannon diversity index was high (0.76). The large number of multilocus genotypes and the presence of both mating types in some fields indicate that sexual reproduction may take place in Estonian populations of P. infestans.     

Characterization of some Asian isolates of Phytophthora infestans

A total of 401 isolates of Phytophthora infestans were collected from eight Asian regions (Korea, India, Taiwan, Indonesia, Thailand, Nepal, China and Japan) between 1992 and 2000 – 318 from potato and 83 from tomato. The isolates were analysed for mating type, metalaxyl resistance, RG57 fingerprinting, mitochondrial DNA (mtDNA) haplotype and the polymorphism of three allozyme loci, i.e. glucose-6-phosphate isomerase ( Gpi ), peptidase ( Pep ) and malic enzyme ( Me ). The isolates were multilocus-genotyped based on RFLP (RG57) fingerprint, dilocus allozyme genotype, mtDNA haplotype and mating type. Twenty multilocus genotypes were identified among 125 isolates. Of these genotypes, 14 had not been previously reported. Some of the multilocus genotypes were common to isolates from several geographical regions, suggesting migration. The metalaxyl-resistant isolates belonged to the multilocus genotypes JP-1, JP-2, and JP-3. Multilocus genotypes coexisting in a single field were found in following regions: Thailand (1994), central China (1996), Nepal (1997) and Japan (1998 and 2000). The possible origins of certain genotypes are discussed, including the possibility of sexual recombination within the P. infestans populations in Nepal and perhaps Thailand.     

Sources of resistance to Pseudomonas syringae pv. phaseolicola races in Phaseolus vulgaris

One thousand and forty-eight Phaseolus bean accessions were evaluated for resistance to six races of Pseudomonas syringae pv. phaseolicola . The accessions originated from regions of the Americas and Africa where the disease is important and included wild type accessions and some known resistance sources. Resistance, graded on a five-point scale, was of two types: qualitative, which was shown to be race-specific, and quantitative. Race specific resistance genes (R-genes) were detected in 49.4% of accessions with the following gene frequencies: R1 (10.3%), R2 (0.3%), R3 (25.0%), R4 (35.0%) and R5 (0.2%).
Evidence for quantitative variation in resistance, in the absence of specific R-genes, was shown by the distribution of infection scores, 76% of accessions showing maximum susceptibility (grades 4–5), 23% showing intermediate resistance (grades 2–4), and 1% showing high levels of quantitative resistance (grades 1–2). The last 1% of accessions showed interactions which were not race-specific and it is suggested that they may possess race non-specific resistance. It is possible that several of the accessions in this category carry the recessive gene derived from PI 150414. Other accessions were of unknown parentage and may represent new sources of quantitative, potentially race non-specific, resistance. It is suggested that the combination of race specific and race non-specific resistance could provide an effective strategy for establishing durable resistance.     

Investigations concerning the role of Chalara fraxinea in declining Fraxinus excelsior

A study was carried out to clarify the role of the fungus Chalara fraxinea in decline of Fraxinus excelsior , which is observed on a large scale in central and northern Europe with high incidence of tree mortality. The aims of this work were: (i) to check for the presence of C. fraxinea in various tissues of declining F. excelsior by agar culture isolations and by direct analysis of plant tissues using molecular techniques (DNA extraction, ITS-PCR, cloning, ITS sequencing and T-RFLP); (ii) to study fungal communities inhabiting tissues with symptoms; and (iii) to test the pathogenicity of C. fraxinea to F. excelsior . Chalara fraxinea was isolated from 93% of stem cankers, 91% of necrotic leaf stalks, 27–28% of bark wounds and 30% of visually healthy leaf stalks. Molecular analyses of necrotic leaves, leaf stalks and bark revealed the presence of 25 different fungal taxa, 14 of which were detected in all three types of tissue sample. Chalara fraxinea was the second most common species (61% of samples), and only Cryptococcus foliicola occurred more often (70%). All eight of the tested C. fraxinea isolates induced necroses in bark and cambium on each of 86 inoculated trees, and all controls remained healthy. Average length of necroses caused by different C. fraxinea strains varied from 4·2 to 8·9 cm, but the differences were statistically insignificant. Instead, differences in resistance of individual trees to C. fraxinea were observed.     

Early molecular signatures of responses of wheat to Zymoseptoria tritici in compatible and incompatible interactions

Zymoseptoria tritici, the causal agent of septoria tritici blotch, a serious foliar disease of wheat, is a necrotrophic pathogen that undergoes a long latent period. Emergence of insensitivity to fungicides, and pesticide reduction policies, mean there is a pressing need to understand septoria and control it through greater varietal resistance. Stb6 and Stb15, the most common qualitative resistance genes in modern wheat cultivars, determine specific resistance to avirulent fungal genotypes following a gene‐for‐gene relationship. This study investigated compatible and incompatible interactions of wheat with Z. tritici using eight combinations of cultivars and isolates, with the aim of identifying molecular responses that could be used as markers for disease resistance during the early, symptomless phase of colonization. The accumulation of TaMPK3 was estimated using western blotting, and the expression of genes implicated in gene‐for‐gene interactions of plants with a wide range of other pathogens was measured by qRT‐PCR during the presymptomatic stages of infection. Production of TaMPK3 and expression of most of the genes responded to inoculation with Z. tritici but varied considerably between experimental replicates. However, there was no significant difference between compatible and incompatible interactions in any of the responses tested. These results demonstrate that the molecular biology of the gene‐for‐gene interaction between wheat and Zymoseptoria is unlike that in many other plant diseases, indicate that environmental conditions may strongly influence early responses of wheat to infection by Z. tritici, and emphasize the importance of including both compatible and incompatible interactions when investigating the biology of this complex pathosystem.     

Pathogenic variation and virulence related responses of <Emphasis Type="Italic">Ascochyta lentis</Emphasis> on lentil

Ascochyta blight of lentil (Lens culinaris ssp. culinaris) is caused by Ascochyta lentis. The disease causes severe damage to all aerial parts of the plant and may lead to total crop loss during extremely severe epidemics. To identify qualitative differences in resistance within Australian lentil crops, variation in virulence was examined among 17 isolates of A. lentis on six differential lentil genotypes (ILL7537, ILL5588 (cv. Northfield), ILL6002, ILL5722 (cv. Digger), ILL481 (cv. Indianhead) and CIPA203 (cv. Nipper)). Six distinct virulence patterns were identified, with Pathotype I (AL4) being highly virulent, causing disease on all genotypes except ILL7537 and pathotype VI (Kewell) exhibiting low virulence on all genotypes. Histopathology studies were carried out to further understand interaction differences between isolate-host combinations and add to the knowledge of possible resistance mechanisms underlying lentil’s defence to the pathogen. The infection process was compared between lentil genotypes with different levels of resistance and isolates with different levels of virulence. Microscopic and biochemical differences were observed between compatible and incompatible interactions, which were related to time-after-inoculation, with slower responses noted in susceptible lentil genotypes. Relatively fast release of reactive oxygen species (ROS) and a subsequent hypersensitive response (HR) was central to initial defence at the point of penetration in the most resistant lentil genotypes.     

The Northern Ireland Phytophthora infestans population 1998–2002 characterized by genotypic and phenotypic markers

A total of 204 isolates of Phytophthora infestans from Northern Ireland, almost all from commercial potato crops, were collected over 5 years (1998–2002). Phenotypic diversity was assessed using mating type and metalaxyl resistance; genotypic diversity was assessed using two allozyme loci (glucose-6-phosphate isomerase, Gpi, and peptidase, Pep ), mitochondrial DNA haplotype and the multilocus RFLP probe RG57. All isolates were A1 mating type and Gpi 100/100 . The majority were Pep 100/100 , but four Pep 83/100 and six Pep 96/100 isolates were identified. Three mtDNA haplotypes were detected; haplotype IIa was the most common, but each year up to 2001 its frequency declined, with a concomitant increase in Ia isolates. Three isolates had the rare haplotype IIb, but this was only detected in 1998. Metalaxyl resistance and mtDNA haplotype were markedly associated: most haplotype Ia isolates were metalaxyl-resistant, whereas haplotype IIa was more commonly associated with metalaxyl sensitivity. Analysis of a subsample of 91 isolates revealed nine RG57 genotypes, three associated exclusively with haplotype IIa and six exclusively with haplotype Ia. The most common RG57 genotype (51% of isolates) comprised both metalaxyl-resistant and -sensitive haplotype IIa isolates. However, of haplotype Ia isolates, all metalaxyl-resistant phenotypes belonged to one of four RG57 types, one of which was the second most frequent overall (29% of isolates), while all metalaxyl-sensitive isolates belonged to one of two other types. The P. infestans population in Northern Ireland appears to consist of a limited number of clones related to, but differentiated from, the populations in mainland Britain and elsewhere in Europe.     

水稻防御反应相关基因对稻瘟病菌的响应

水稻与稻瘟病菌的互作已成为研究植物与病原真菌互作的模式系统。利用RT-PCR技术检测了6个水稻品种(分别含有抗病基因Pik-s、Pita、Pit、Pi1、Pi9的近等系及回交亲本丽江新团黑谷LTH)与稻瘟病菌互作过程中多个信号相关及PR基因的表达。结果表明带有稻瘟病抗性基因Pik-s、Pita、Pit的水稻品种和LTH对稻瘟病菌#626侵染表现为亲和互作,带Pi1和Pi9的水稻品种表现为非亲和互作;稻瘟病菌接种后,亲和互作中MAPK6和MAPK12表现为上调表达,带有抗性基因Pi9的水稻品种IRBL22中BIMK2表现为上调表达。总体来看,含有不同抗病基因的水稻近等系中的PR基因对稻瘟病菌的响应较为多样,非亲和互作中在早期或早中期表现为PR基因上调表达,而亲和互作中主要在晚期上调表达,说明这些PR基因表达的时间在植物与病原互作的不同时期发挥着不同的作用。     

An optimized screening method for identifying levels of resistance to Crinipellis perniciosa in cocoa (Theobroma cacao)

The effects of host age, leaf number, host type (clone or seedling), pathogen spore concentration and incubation time on inoculation with Crinipellis perniciosa (witches' broom disease of cocoa) were studied in greenhouse experiments using susceptible cocoa genotypes. Three methods of inoculation (agar-drop, water-drop and spray) were also tested. An optimized inoculation method was selected and tested for its repeatability as well as its ability to discriminate between various levels of resistance to C. perniciosa in cocoa. The optimized method (350 000 viable basidiospores per mL, 60 h incubation, agar-drop technique) produced 100% infection repeatedly, on both clonal and seedling plants of a susceptible genotype. Seedling age (2–12 months) and leaf number did not significantly affect the percentage of plants with symptoms or broom characteristics. This method discriminated effectively between the various levels of resistance in 14 cocoa genotypes and is recommended as an inoculation method to identify levels of resistance in germplasm collections. Symptom severity was shown to be a better measure of resistance than infection success.     

Identification of new resistant sources for bacterial blight in pomegranate

Bacterial blight is a highly devastating disease caused by Xanthomonas axonopodis pv. punicae, recording 60 to 80 percent yield-loss of pomegranate in India. In the present investigation, a total of 209 genotypes including 105 exotic types from USDA, 66 wild types and 38 cultivated types from India were screened and categorized into fifteen clusters using cluster and principal component analysis. Genotypes of cluster 15, viz. 108 B and 99 A from USDA and 318734, Daru-18 and IIHR-30 from India, were found to be resistant to bacterial blight while genotypes of cluster 9 were highly susceptible. Two genotypes, each from cluster 15 (318734) and 9 (Ruby), were compared for biochemical and histological parameters to understand the defense mechanism. Significantly, higher accumulation of defense related metabolites, viz. total phenol, flavonoid and antioxidant contents, were observed in resistant genotype (318734). Fewer numbers of stomatal pores that served as portals of entry for plant pathogens were recorded in this genotype. Resistance observed in genotype 318734 might be due to an incompatible interaction between host and pathogen compared to other genotypes. This is the first report of putative resistance sources in pomegranate against Xanthomonas axonopodis pv. punicae.     

New Avirulence Genes in the Phytopathogenic Fungus Leptosphaeria maculans

ABSTRACT Leptosphaeria maculans, the causal agent of stem canker of oilseed rape (Brassica napus), develops gene-for-gene interactions with oilseed rape, and four L. maculans avirulence (AVR) genes (AvrLm1, AvrLm2, AvrLm4, and alm1) were previously genetically characterized. Based on the analysis of progeny of numerous in vitro crosses between L. maculans isolates showing either already characterized or new differential interactions, this work aims to provide an overview of the AVR genes that may specify incompatibility toward B. napus and the related species B. juncea and B. rapa. Two novel differential interactions were thus identified between L. maculans and B. napus genotypes, one of them corresponding to a complete resistance to European races of L. maculans. In both cases, a single gene control of avirulence was established (genes AvrLm3 and AvrLm7). Similarly, a single gene control of avirulence toward a B. rapa genotype, also resistant to European L. maculans isolates, was demonstrated (gene AvrLm8). Finally, a digenic control of avirulence toward B. juncea was established (genes AvrLm5 and AvrLm6). Linkage analyses demonstrated that at least four unlinked L. maculans genomic regions, including at least one AVR gene cluster (AvrLm1-AvrLm2-AvrLm6), are involved in host specificity. The AvrLm3-AvrLm4-AvrLm7 region may correspond either to a second AVR gene cluster or to a multiallelic AVR gene.     

小麦与秆锈菌非亲和性互作对秆锈菌生长发育的影响

 本文研究了小麦与秆锈菌3种典型类型非亲和性互作对秆锈菌生长发育的影响。结果表明,完全非亲和性互作ISr5-Ra/21C₃ CKR(Sr5/P5)使秆锈菌芽管生长方向与叶片长轴夹角显著变小,比对应的亲和性互作ISr5-Sa/21C₃ CKR(sr5/p5)平均小6.22°。高度非亲和性互作ISr 11-Ra/34M KG(Sr11/P11)使秆锈菌附着胞总分化率显著降低,接种后第1、2 d平均分别降低21.16%、33.37%,3种类型非亲和性互作对附着胞在气孔上的定位无影响。各类非亲和性互作对秆锈菌菌落扩展有不同程度极显著抑制作用。完全非亲和性互作、高度非亲和性互作及中度非亲和性互作ISr 6-Ra/34C2 MFR(Sr6/P6)侵染点寄主细胞坏死率分别为66.7%~76%、近100%及52.4%~87.1%。     

Changes in specific virulence in Polish populations ofPhytophthora infestans: 1985–1991

Ninety-five isolates ofPhytophthora infestans collected throughout Poland during 1985–1991 and characterized for multilocus genotypes based on mating type, allozymes and DNA fingerprint, were analyzed for specific virulence to differential potato cultivars carrying ten major resistance genes. The multilocus analysis led to three groupings. The first group contained 22 isolates of a clonal lineage (PO-1) that is postulated to have been present in Europe during most of the twentieth century, but PO-1 isolates were recovered in Poland only during 1985–1988. This group contained, on average, virulence to 5.5 specific resistance genes per isolate. The second group consisted of 30 isolates in a clonal lineage (PO-4) that had not been detected before 1988. PO-4 isolates had virulence to a mean of 6.5 resistance genes per isolate. The third group was composed of 43 isolates representing 38 multilocus genotypes also not detected before 1988. These diverse genotypes had virulence to an average of 6.7 specific resistance genes per isolate. More than half (53%) of the PO-4 isolates shared a single pathotype. The group of 43 isolates was dominated by two pathotypes: the most common one (47% of the isolates) was the same pathotype that dominated PO-4 isolates; the next most common one (21%) differed from the most common one by the absence of virulence to resistance gene R5. The recent immigrant isolates (not detected before 1988) generally had virulence to a greater number of specific resistance genes than did isolates in the previous population [detected before 1988 (PO-1)]. Recent immigrant populations were dominated by one or two pathotypes, so their pathotypic diversity values were somewhat less than that of the previous population.     

Systemic acquired resistance delays race shifts to major resistance genes in bell pepper

ABSTRACT The lack of durability of host plant disease resistance is a major problem in disease control. Genotype-specific resistance that involves major resistance (R) genes is especially prone to failure. The compatible (i.e., disease) host-pathogen interaction with systemic acquired resistance (SAR) has been studied extensively, but the incompatible (i.e., resistant) interaction less so. Using the pepper-bacterial spot (causal agent, Xanthomonas axonopodis pv. vesicatoria) pathosystem, we examined the effect of SAR in reducing the occurrence of race-change mutants that defeat R genes in laboratory, greenhouse, and field experiments. Pepper plants carrying one or more R genes were sprayed with the plant defense activator acibenzolar-S-methyl (ASM) and challenged with incompatible strains of the pathogen. In the greenhouse, disease lesions first were observed 3 weeks after inoculation. ASM-treated plants carrying a major R gene had significantly fewer lesions caused by both the incompatible (i.e., hypersensitive) and compatible (i.e., disease) responses than occurred on nonsprayed plants. Bacteria isolated from the disease lesions were confirmed to be race-change mutants. In field experiments, there was a delay in the detection of race-change mutants and a reduction in disease severity. Decreased disease severity was associated with a reduction in the number of race-change mutants and the suppression of disease caused by the race-change mutants. This suggests a possible mechanism related to a decrease in the pathogen population size, which subsequently reduces the number of race-change mutants for the selection pressure of R genes. Thus, inducers of SAR are potentially useful for increasing the durability of genotype-specific resistance conferred by major R genes.     

Interaction of common bacterial blight bacteria with disease resistance quantitative trait loci in common bean

Common bacterial blight (CBB) of common bean (Phaseolus vulgaris L.) is caused by Xanthomonas campestris pv. phaseoli and X. fuscans subsp. fuscans, and is the most important bacterial disease of this crop in many regions of the world. In 2005 and 2006, dark red kidney bean fields in a major bean-growing region in central Wisconsin were surveyed for CBB incidence and representative symptomatic leaves collected. Xanthomonad-like bacteria were isolated from these leaves and characterized based upon phenotypic (colony) characteristics, pathogenicity on common bean, polymerase chain reaction (PCR) with X. campestris pv. phaseoli- and X. fuscans subsp. fuscans-specific primers, and repetitive-element PCR (rep-PCR) and 16S-28S ribosomal RNA spacer region sequence analyses. Of 348 isolates that were characterized, 293 were identified as common blight bacteria (i.e., pathogenic on common bean and positive in PCR tests with the X. campestris pv. phaseoli- and X. fuscans subsp. fuscans-specific primers), whereas the other isolates were nonpathogenic xanthomonads. Most (98%) of the pathogenic xanthomonads were X. campestris pv. phaseoli, consistent with the association of this bacterium with CBB in large-seeded bean cultivars of the Andean gene pool. Two types of X. campestris pv. phaseoli were involved with CBB in this region: typical X. campestris pv. phaseoli (P) isolates with yellow mucoid colonies, no brown pigment production, and a typical X. campestris pv. phaseoli rep-PCR fingerprint (60% of strains); and a new phenotype and genotype (Px) with an X. campestris pv. phaseoli-type fingerprint and less mucoid colonies that produced brown pigment (40% of strains). In addition, a small number of X. fuscans subsp. fuscans strains, representing a new genotype (FH), were isolated from two fields in 2005. Representative P and Px X. campestris pv. phaseoli strains, an FH X. fuscans subsp. fuscans strain, plus five previously characterized X. campestris pv. phaseoli and X. fuscans subsp. fuscans genotypes were inoculated onto 28 common bean genotypes having various combinations of known CBB resistance quantitative trait loci (QTL) and associated sequence-characterized amplified region markers. Different levels of virulence were observed for X. campestris pv. phaseoli strains, whereas X. fuscans subsp. fuscans strains were similar in virulence. The typical X. campestris pv. phaseoli strain from Wisconsin was most virulent, whereas X. campestris pv. phaseoli genotypes from East Africa were the least virulent. Host genotypes having the SU91 marker-associated resistance and one or more other QTL (i.e., pyramided resistance), such as the VAX lines, were highly resistant to all genotypes of common blight bacteria tested. This information will help in the development of CBB resistance-breeding strategies for different common bean market classes in different geographical regions, as well as the identification of appropriate pathogen genotypes for screening for resistance.