Traditional and commercial apple and pear cultivars as sources of resistance to fireblight1

In Hungary, fireblight research programmes were initiated on pear in 1999 and on apple in 2000, with the aim of evaluating the susceptibility or resistance of commercial cultivars. Sources of resistance for future breeding were also sought among traditional apple cultivars collected from Ukraine and pear cultivars in the Hungarian gene bank (Szigetcsép). Experiments were done under secure conditions. Inocula were mixtures of characteristic Erwinia amylovora isolates from pear and apple in Hungary. Host responses (symptom development, disease severity and multiplication rate of bacterial cells in host tissues) were assessed on shoots, flowers and fruits. About 30 pear and 30 apple cultivars, and 35 apple hybrids, were tested and grouped into four categories for pear and three for apple. Of the pear cultivars tested, 50% were susceptible, 30% moderately susceptible and only 10% of low susceptibility. Different plant organs occasionally displayed different responses. Members of the last two groups might serve as useful candidates for growing under IPM conditions. Among the traditional Hungarian varieties tested, we found high resistance in ‘Sikulai’ and ‘Szemes alma’, which could be used as sources of fireblight resistance in breeding programmes and also grown in organic orchards. Furthermore, among the offspring of the apple ‘Prima’ (scab‐resistant), we have found highly resistant lines.     

Resistance and Resistance Breeding in Ornamentals 1

Heritability studies in ornamentals with regard to Erwinia amylovora (Burrill) Winslow et al. were started shortly after the introduction of the pathogen into northern Europe. Since that time, approximately 8,000 plants from 17 genera of Rosaceae have been tested; 7 genera were reported to be susceptible: the economically important Cotoneaster, Crataegus, Cydonia, Pyracantha, Sorbus, and, in addition, Chaenomeles and Stranvaesia. Moreover, 9,000 Cotoneaster seedlings comprising 15 cultivars were tested and the surviving individuals used for breeding purposes. The variation between these first results reported in the different European countries and some American findings is considerable because of different inoculation procedures and various epidemiological conditions. However, resistance sources have been identified and it would be desirable to start co–operation in this field on an international level.     

Effectiveness of the resistance inducer prohexadione‐Ca against fireblight in shoots of apple trees inoculated with Erwinia amylovora1

Due to the lack of effective, non‐phytotoxic and publicly acceptable materials for controlling fireblight in pome fruit trees, novel strategies against Erwinia amylovora are being sought. Resistance‐inducing compounds, such as prohexadione‐Ca, represent promising alternatives. Prohexadione‐Ca is the active substance of the bioregulator Regalis, currently being introduced in several European countries and overseas. Prohexadione‐Ca reduces shoot elongation due to inhibition of gibberellin biosynthesis. Furthermore, it leads to significant changes in the spectrum of flavonoids and their phenolic precursors in pome fruits, which causes reduced susceptibility to fireblight and other pathogens. In 2002 and 2003, container‐grown apple trees of the cultivars ‘Idared’ and ‘Freedom’ were treated with different dosages of prohexadione‐Ca two weeks before inoculation with E. amylovora. The effect of prohexadione‐Ca against shoot blight was determined by measuring the lengths of necrotic lesions and symptoms on vascular bundles caused by the pathogen. Treatments with prohexadione‐Ca turned out to be much superior to the ones with streptomycin, kasugamycin and a bacterial antagonist, which were used for comparison. Acibenzolar‐S‐methyl (Bion), another resistance‐inducing compound, was included in some of the experiments and gave intermediate results. The simultaneous control of excessive shoot growth and shoot infections by fireblight is seen as a major advantage of using prohexadione‐Ca in pome fruit trees.     

Official survey,in 2002, for detection of Erwinia amylovora in a proposed protected zone in Jihomoravsky region (Czech Republic)1

Inspectors of the Czech NPPO surveyed the occurrence of fireblight Erwinia amylovora in an area of Jihomoravsky region (South Moravia) proposed as an EU protected zone, including 16 designated buffer zones around nurseries. The disease was not detected in communes where fireblight hosts are grown (nurseries, variety testing stations, orchards) or in buffer zones around nurseries. In 902 communes where fireblight hosts are grown only in orchards or not at all, wild host plants were inspected at 2.629 observation points (2137 located by GARMIN GPS). In Vy?kov district, suspected fireblight was confirmed at one observation point on Crataegus, growing by a railway in Rousínov commune and, in the course of a delimiting survey outside observation points, in four other communes (Drnovice, Habrovany, Komo?any and Vy?kov).     

Tomato early blight (Alternaria solani): the pathogen, genetics, and breeding for resistance

Alternaria solani causes diseases on foliage (early blight), basal stems of seedlings (collar rot), stems of adult plants (stem lesions), and fruits (fruit rot) of tomato. Early blight is the most destructive of these diseases and hence receives considerable attention in breeding. For over 60 years, breeding for early blight resistance has been practiced, but the development of cultivars with high levels of resistance has been hampered by the lack of sources of strong resistance in the cultivated tomato and by the quantitative expression and polygenic inheritance of the resistance. In some accessions of wild species, high levels of early blight resistance have been found, but breeding lines still have unfavorable horticultural traits from the donor parent. Recently, the first linkage maps with loci controlling early blight resistance have been developed based on interspecific crosses. These maps may facilitate marker-assisted selection. This overview presents the current knowledge about the A. solani–tomato complex with respect to its biology, genetics, and breeding.     

Novel metrics to classify fire blight resistance of 94 apple cultivars

Fire blight (Erwinia amylovora), a potentially devastating disease in apple, can cause floral, fruit and structural damage and even tree death. Most commercial apple cultivars are susceptible and the resistance/susceptibility of many modern cultivars has not been evaluated. Fire blight resistance/susceptibility is difficult to phenotype due to quantitative resistance, impacts of tree vigour and environment on susceptibility, and the erratic nature of the disease. Resistance/susceptibility levels were determined for 94 apple cultivars and important breeding parents. In 2016 and 2017, multiple actively growing shoots per tree (about three trees per cultivar) were challenged with E. amylovora Ea153n via a cut-leaf inoculation method. Proportion of current season's shoot length blighted (SLB) was calculated for each shoot. To classify cultivar responses, estimated marginal SLB means were compared to four controls, representing highly susceptible (HS) to highly resistant (HR), via Dunnett's tests. Cultivar responses ranged from HS to HR with estimated marginal SLB means of 0.001–0.995 in 2016 and 0.000–0.885 in 2017. Most cultivars demonstrated similar resistance/susceptibility levels in both years (ρ = 0.657, P < 0.0001). K-means clustering was used to classify cultivars into three resistance/susceptibility groups based on incidence, average severity (SLB), and maximum severity values (maximum SLB and age of wood infected). Sixteen cultivars were consistently moderately resistant (MR) to HR while the remainder ranged from HS to MR. An updated comparison of susceptibility of important cultivars is provided. Resistance/susceptibility information gained could be used to identify genetic loci associated with resistance/susceptibility and/or inform parental selection in apple scion breeding programmes.     

Distribution of streptomycin-resistant strainsof Erwinia amylovora in Israel and occurrence of blossom blight in the autumn

Following failure in control of fire blight with streptomycin, the distribution of streptomycin-resistant strains ofErwinia amylovora in Israel was surveyed. During 1994–1997 109 pear, apple, loquat and quince orchards were monitored. Streptomycin-resistant strains ofE. amylovora were recovered from flowers and from infected branches collected at 18 locations in the Sharon, Galilee and Golan Heights regions. In the Sharon region all the isolated strains ofE. amylovora were streptomycin-resistant, whereas in the Galilee and Golan Heights, resistant as well as sensitiveE. amylovora strains were recovered at different locations. In the southern coastal plain no resistance could be detected. Streptomycin-resistant strains ofE. amylovora did not hybridize with the DNA probe SMP3, and resistance could not be transferred by mating to a sensitive strain, suggesting that streptomycin resistance in Israel is not plasmid-mediated. Fire blight symptoms were observed, for the first time, on pear blossoms during the autumn of 1994. A high population of 2x 10⁶-6x 10⁷ CFU/flower in the autumn of 1995 and of 1996 was correlated with the appearance of blossom blight symptoms.     

Experience with fireblight in the Czech Republic1

Erwinia amylovora has been spreading in the Czech Republic since 1986 regardless of emergency phytosanitary measures which have been taken. Its spread follows the predominant wind direction (west to east). The infested area now covers two thirds of the area of the country. Wild Crataegus spp. are the most important and widespread host plants. E. amylovora is still regulated as a quarantine pest, and areas free from fireblight are subject to special phytosanitary measures.     

Fireblight Inoculum: Sources and Dissemination1

Understanding the disease cycle of fireblight requires knowledge of the sources of inoculum and means of dissemination. In regions where the disease is endemic, hold–over cankers are the most important source of primary inoculum. Dissemination of primary inoculum occurs by rain or by crawling or flying insects. Often, blossoms infested with Erwinia atnylovora (Burrill) Winslow et al. provide secondary inoculum that is disseminated by pollinating insects or by rain. Later in the growing season, secondary inoculum produced in infected tissues may be disseminated by rain or by sucking and biting insects. Relatively rare sources of inoculum and inefficient means of dissemination may be responsible for introducing the pathogen to areas where fireblight has not been found previously. E. amylovora may be carried to new locations in intimate association with host tissues (infected nursery stock), in association with vectors (insects, birds or man), or by wind as “strands” or in airborne water drops. Every effort should be made to prevent introduction of E. amylovora into areas now thought to be free of fireblight. Attempts to eradicate the disease from several new areas in northern Europe have been unsuccessful. Because weather conditions and the host plants present in southern Europe are especially conducive to severe damage from fireblight, plant protection personnel are advised to prepare now for the possible introduction of disease. Personnel should be trained to recognize the disease and plans should be drawn to contain it or to manage host plants to minimize disease losses.     

Fireblight in Békés County (Hungary) in 1996/20021

Fireblight (Erwinia amylovora) appeared in Hungary in 1996. Most damage occurred on apple, pear, quince and medlar, and also on the ornamentals Pyracantha, Sorbus, Cotoneaster and Crataegus. In 1996–2006, an official programme for elimination of infected parts of plants started in Békés county. This mainly concerned trees in towns and villages, since there are few pome‐fruit orchards in the county. Work teams under official direction pruned back or cut down trees. In total, some 13 000 trees were pruned back and nearly 11 000 were cut down. In addition, 21 villages were subjected to special phytosanitary measures. Infection decreased considerably between 1996 and 2002, but over 90% of the inhabited areas in the county remained subject to special measures, because of the very dispersed occurrence of fireblight.     

Variability and selection of verticillium wilt resistant genotypes in cultivated olive and in the Olea genus

Developing verticillium wilt resistant genotypes is currently a major objective in olive breeding. In this study, 6017 genotypes derived from 48 crosses obtained by open pollination and crosses between olive cultivars, wild olive genotypes and other Olea species and Olea europaea subspecies were individually evaluated for verticillium wilt resistance. More than 800 genotypes were identified as resistant to the disease based on the absence of symptoms. High genetic variability and wide segregation in resistance were observed. The inheritance of resistance was studied, and the best parents and crosses to breed resistant genotypes were identified. According to the results, verticillium wilt resistance in olive appears to be a quantitative trait. The results obtained by comparing the level of resistance between different crosses as well as by estimating heritability suggest that it is possible to breed for verticillium wilt resistance in olive.     

Protective Effects Against Erwinia amylovora Induced by hrp Mutants in the Whole Plant are Only Partially Mimicked in Cultivated Cells

A virulent strain of Erwinia amylovora, the causal agent of fire blight of Maloideae, and two of its non-virulent hrp mutants (a secretory and a regulatory mutant) were inoculated into apple cell suspensions either alone or in mixed inoculations. In single inoculations, death of 4- to 5-day-old apple cells occurred only when the concentration of the virulent strain of E. amylovora reached a threshold inoculum concentration of 10⁴CFUml^–1, while high concentrations of the hrp mutants were unable to kill apple cells. When mixed inoculated with the virulent parental strain, both hrp mutants protected apple cells from death caused by the virulent strain. The protective effect was associated with a decrease in the population level of the virulent strain and it was dependent on the non-virulent to virulent inoculum concentration suggesting a competition between the non-virulent mutant and the virulent strain. However, no differential protective ability between the two types of mutants could be noticed, contrary to previous results obtained with apple seedlings or apple flowers in which the regulatory mutant was significantly more effective than the secretory mutant. In conclusion, inoculation of apple cell cultures with E. amylovora does not seem to be a model suitable for investigating mechanisms leading to protection.     

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.     

Identification of avirulence genes in the rice blast fungus corresponding to three resistance genes in Japanese differentials

The pathogenicity of progeny from crosses among three Chinese isolates of Magnaporthe grisea collected from rice was tested on three Japanese differentials (Ishikarishiroke, Aichiasahi, K 59) having the blast resistance genes Pii, Pia, and Pit, respectively. Monogenic control was demonstrated for avirulence to the differentials. To identify resistance genes corresponding to the avirulence genes, the resistance and susceptibility in F₃ lines of the cultivars in response to the parents of the crosses were analyzed genetically. The three avirulence genes identified, designated Avr-Pii, Avr-Pia, and Avr-Pit, appear to correspond to resistance genes Pii, Pia, and Pit, respectively. The monogenic control of avirulence in the fungus and monogenic dominant resistance in rice cultivars supports a gene-for-gene relation in the Pii-, Pia-, or Pit-dependent resistance to the rice blast fungus in rice cultivars.     

Significance of PWT4–Rwt4 interaction in the species specificity of Avena isolates of Magnaporthe oryzae on wheat

We examined whether PWT4, an avirulence gene of Avena isolates of Magnaporthe oryzae toward wheat, corresponded to Rwt4, a resistance gene identified in wheat cultivar Norin 4, in a one-to-one manner. Twelve wheat cultivars were inoculated with 65X1, an F₁ culture with PWT4 derived from a cross between an Avena isolate (Br58) and a Triticum isolate (Br48). Three wheat cultivars (Norin 26, Shin-chunaga, Cheyenne) were resistant and therefore selected as possible carriers of Rwt4. The three cultivars were then inoculated with a population derived from a backcross of 61M2 carrying PWT4 with Br48 carrying pwt4. Segregation analyses revealed that PWT4 operates against the three cultivars. If PWT4 corresponds to Rwt4 in a one-to-one manner, all three cultivars should carry Rwt4. To test if this is the case, the three cultivars were crossed with Chinese Spring (a noncarrier of Rwt4) and Norin 4. When F₂ seedlings from Chinese Spring × Norin 26, Chinese Spring × Shin-chunaga, and Chinese Spring × Cheyenne were inoculated with 61M2, resistant and susceptible seedlings segregated in a 3 : 1 ratio. On the other hand, crosses between the three cultivars and Norin 4 yielded no susceptible F₂ seedlings. These results indicate that all three cultivars carry Rwt4. Considering all results, we concluded that PWT4 corresponds to Rwt4 in a one-to-one manner. An inoculation test with Chinese Spring–Cheyenne chromosome substitution lines indicated that Rwt4 is located on chromosome 1D.     

Evaluation Of Tests to Determine Resistance of Zantedeschia spp. (Araceae) to Soft Rot caused by Erwinia carotovora subsp. carotovora

Bacterial soft rot caused by Erwinia carotovora subsp. carotovora is a major disease in Zantedeschia spp., particularly in cultivars from the section Aestivae. The disease can be partly controlled by cultural measures, but by combining cultural methods with resistant plant material a promising strategy for control of soft rot can be developed. No tests are available for resistance testing in breeding Zantedeschia spp. Therefore, three tests developed for use in potato breeding were adapted for use on eight cultivars of Zantedeschia spp. Variation was found in all three tests. Resistant control cultivar Zantedeschia aethiopica Crowborough scored most resistant in all three tests. Within the section Aestivae, degrees of susceptibility were identified that were in agreement with each other and with field observations, indicating reliability of two of the methods in which tubers were used. The correlation coefficient of these two tests was high. A new non-destructive test method was developed for use on seedlings which involved immersion of leaf disks in a bacterial suspension. The percentage of decayed leaf area was a measure of resistance and results were in general agreement with the other tests. These methods will be useful for breeding for soft rot resistance and performing genetic analyses.     

Molecular and biological characterization of two potyviruses infecting lettuce in southeastern France

Several potyviruses affect lettuce (Lactuca sativa) and chicory (Cichorium spp.) crops worldwide and are important constraints for production because of the direct losses that they induce and/or because of their seed transmission. Here, the molecular and biological properties are described of two potyviruses that were recently isolated from lettuce plants showing mosaic or strong necrotic symptoms in an experimental field in southeastern France. The first potyvirus belongs to the species Endive necrotic mosaic virus and is present in a large number of wild plant species, especially Tragopogon pratensis. It is unable to infect lettuce cultivars with a resistance to Turnip mosaic virus that is present in many European cultivars and probably conferred by the Tu gene. The second potyvirus belongs to the tentative species lettuce Italian necrotic virus and was not observed in wild plants. It infected all tested lettuce cultivars. Wild accessions of Lactuca serriola, Lactuca saligna, Lactuca virosa and Lactuca perennis were identified as resistant to one or the other potyvirus and could be used for resistance breeding in lettuce. No resistance against these two potyviruses was observed in the tested Cichorium endivia cultivars. In contrast, all tested Cichorium intybus cultivars or accessions were resistant.     

Occurrence of bacterial black shoot disease of European pear in Yamagata Prefecture

Black lesions on shoots of European pear trees observed in an orchard in Yamagata Prefecture in May 2007 were suspected to be caused by a bacterial pathogen. The surface of the colonies isolated on a high sucrose medium did not have the crater morphology that is characteristic of E. amylovora bvs. 1–3, and a specific DNA fragment was amplified from the isolates in the PCR using the EprpoD primer set. The partial sequences of the 16S rRNA gene placed the isolates in the genus Erwinia. The isolates differed serologically from E. amylovora biovars and E. pyrifoliae in an Ouchterlony double-diffusion test although their bacterial properties suggested that they are closely related to E. amylovora biovars and E. pyrifoliae. In a DNA–DNA hybridization test, the relatedness between the isolates and E. amylovora biovars or E. pyrifoliae did not exceed 70% level, indicating that they are independent species. Thus, the isolates belongs to the genus Erwnia but are not E. amylovra or E. pyrifoliae. After succulent pear shoots were injected with bacterial suspensions (10⁹, 10⁸, 10⁷ and 10⁶ cfu/ml) of the isolates, lesions formed with 10⁹ and 10⁸ cfu/ml, but the disease incidence with 10⁸ cfu/ml was much lower than with E. amylovora and E. pyrifoliae. Virulence of the present isolates is thus thought to be very weak. On the basis of these results, we consider that this is a new shoot disease of European pear. In the 2007 season, all affected trees were pulled out after harvest. No symptoms have been observed in field surveys since the fruitlet season in 2007.     

Comparing rice germplasm groups for growth, grain yield and weed-suppressive ability under aerobic soil conditions

Germplasm and cultivars need to be selected as parents for breeding weed‐competitive aerobic rice in the tropics. Forty rice (Oryza sativa) cultivars belonging to the aus, indica and tropical japonica germplasm groups, or derived from crosses among them, were evaluated in adjacent weed‐free and weedy trials in aerobic soil conditions during the wet seasons of 2001–2003. The objectives of this study were to assess vegetative growth, grain yield under weed‐free (Y_F) and weedy (Y_W) conditions, and weed‐suppressive ability (WSA) of different germplasm groups. In the first 4 weeks after sowing, indica cultivars had faster growth in height, tillering and crop biomass than other groups. They also had high Y_F, Y_W and strong WSA. Aus cultivars were similar to the indica types in early growth and WSA, but were poor in Y_F. Tropical japonica groups and the group derived from indica/tropical japonica crosses were generally inferior to aus and indica groups in early growth and WSA. Both of their Y_F and Y_W were lower than that of the indica group. Therefore, indica germplasm seemed to be most suitable for breeding high‐yielding and weed‐suppressive aerobic rice for the tropics. The relationship of WSA with various traits within tropical japonica germplasm revealed that fast early growth, rather than plant erectness, is crucial to WSA.     

Spot blotch disease of wheat: the current status of research on genetics and breeding

The spot blotch disease of wheat is caused by Bipolaris sorokiniana, which is an anamorph (teleomorph Cochliobolus sativus). The disease mainly occurs in warm, humid wheat‐growing regions, and the Eastern Gangetic Plains (EGP) of South Asia is a hotspot. Significant progress has been made in recent years in characterizing the host–pathogen interaction. The study of the pathogen's life cycle and diversity have been an active area of research. A number of resistance sources have also been identified, characterized and used for breeding. Although immunity has not been observed in any genotype, cultivars displaying a relatively high level of resistance have been developed and made available to farmers. Further progress will require regular use of marker‐assisted breeding, genomic selection, gene editing and transgenic interventions. This review summarizes the current state of knowledge about genetic and breeding efforts on the wheat–B. sorokiniana pathosystem and discusses ways in which emerging tools can be used for future research to understand the mechanism involved in infection and for developing cultivars exhibiting a high level of resistance.