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Potato ( Solanum tuberosum L.) ranks fourth among the most important staple food in the world. Ralstonia solanacearum (phylotype [phy] IIB, sequevar [seq] 1 and 2), also known as R3B2, the causal agent of brown rot disease on potato, is extremely damaging, causing great economical losses to potato in temperate regions. It is thought that members of Ralstonia pseudosolanacearum (phy I) are not pathogenic at low temperatures and are usually found in warmer climates. R. pseudosolanacearum strain PD 7123 (seq 33) isolated from roses in the Netherlands, strain P824 (seq 13) isolated from blueberry, and strain P781 (seq 14) from mandevilla in Florida are phylogenetically closely related and could share the same host. The virulence and ability of these novel strains to multiply latently in potato in temperate regions is unknown. The objective of this work was to assess the virulence and presence of latent infections of the mentioned R. pseudosolanacearum strains on three commercial seed potato cultivars under warmer (28°C) and temperate (20°C) temperatures. At 28°C, all three R. pseudosolanacearum strains caused severe symptoms on all potato cultivars. Overall disease severity on potato was lower at 20°C than 28°C, but major differences in virulence of the three strains were observed at 42 days postinoculation (dpi) among potato cultivars. All asymptomatic potato plants and most of their daughter tubers had latent infections at 20°C. Altogether, these results show that the phy I strains from rose, blueberry, and mandevilla may pose a threat to potato production in temperate climates and the worldwide movement of seed potatoes.[Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY 4.0 International license., Competing Interests: The author(s) declare no conflict of interest.

Key Message: Rose has 19 MLO genes. Of these, RhMLO1 and RhMLO2 were shown to be required for powdery mildew infection, which suggests their potential as susceptibility targets towards disease resistance. Powdery mildew, caused by Podosphaera pannosa, is one of the most serious and widespread fungal diseases for roses, especially in greenhouse-grown cut roses. It has been shown that certain MLO genes are involved in powdery mildew susceptibility and that loss of function in these genes in various crops leads to broad-spectrum, long-lasting resistance against this fungal disease. For this reason, these MLO genes are called susceptibility genes. We carried out a genome-wide identification of the MLO gene family in the Rosa chinensis genome, and screened for allelic variants among 22 accessions from seven different Rosa species using re-sequencing and transcriptome data. We identified 19 MLO genes in rose, of which four are candidate genes for functional homologs in clade V, which is the clade containing all dicot MLO susceptibility genes. We detected a total of 198 different allelic variants in the set of Rosa species and accessions, corresponding to 5-15 different alleles for each of the genes. Some diploid Rosa species shared alleles with tetraploid rose cultivars, consistent with the notion that diploid species have contributed to the formation of tetraploid roses. Among the four RhMLO genes in clade V, we demonstrated using expression study, virus-induced gene silencing as well as transient RNAi silencing that two of them, RhMLO1 and RhMLO2, are required for infection by P. pannosa and suggest their potential as susceptibility targets for powdery mildew resistance breeding in rose., (© 2021. The Author(s).)

Replant disease is a worldwide phenomenon affecting various woody plant genera and species, especially within the Rosaceae. Compared to decades of intensive studies regarding replant disease of apple (ARD), the replant disease of roses (RRD) has hardly been investigated. The etiology of RRD is also still unclear and a remedy desperately needed. In greenhouse pot trials with seedlings of the RRD-sensitive rootstock Rosa corymbifera 'Laxa' cultured in replant disease affected soils from two different locations, early RRD symptom development was studied in fine roots. In microscopic analyses we found similarities to ARD symptoms with regards to structural damages, impairment in the root hair status, and necroses and blackening in the cortex tissue. Examinations of both whole mounts and thin sections of fine root segments revealed frequent conspicuous fungal infections in association with the cellular disorders. Particularly striking were fungal intracellular structures with pathogenic characteristics that are described for the first time. Isolated fungi from these tissue areas were identified by means of ITS primers, and many of them were members of the Nectriaceae. In a next step, 35 of these isolates were subjected to a multi-locus sequence analysis and the results revealed that several genera and species were involved in the development of RRD within a single rose plant. Inoculations with selected single isolates (Rugonectria rugulosa and Ilyonectria robusta) in a Perlite assay confirmed their pathogenic relationship to early necrotic host plant reactions, and symptoms were similar to those exhibited in ARD.

Key Message: Two environmentally stable QTLs linked to black spot disease resistance in the Rosa wichurana genetic background were detected, in different connected populations, on linkage groups 3 and 5. Co-localization between R-genes and defense response genes was revealed via meta-analysis. The widespread rose black spot disease (BSD) caused by the hemibiotrophic fungus Diplocarpon rosae Wolf. is efficiently controlled with fungicides. However, in the actual context of reducing agrochemical use, the demand for rose bushes with higher levels of resistance has increased. Qualitative resistance conferred by major genes (Rdr genes) has been widely studied but quantitative resistance to BSD requires further investigation. In this study, segregating populations connected through the BSD resistant Rosa wichurana male parent were phenotyped for disease resistance over several years and locations. A pseudo-testcross approach was used, resulting in six parental maps across three populations. A total of 45 individual QTLs with significant effect on BSD resistance were mapped on the male maps (on linkage groups (LG) B3, B4, B5 and B6), and 12 on the female maps (on LG A1, A2, A3, A4 and A5). Two major regions linked to BSD resistance were identified on LG B3 and B5 of the male maps and were integrated into a consensus map built from all three of the male maps. A meta-analysis was used to narrow down the confidence intervals of individual QTLs from three populations by generating meta-QTLs. Two 'hot spots' or meta-QTLs were found per LG, enabling reduction of the confidence interval to 10.42 cM for B3 and 11.47 cM for B5. An expert annotation of NBS-LRR encoding genes of the genome assembly of Hibrand et al. was performed and used to explore potential co-localization with R-genes. Co-localization with defense response genes was also investigated.

Background: The AP2/ERFs belong to a large family of transcription factors in plants. The AP2/ERF gene family has been identified as a key player involved in both biotic and abiotic stress responses in plants, however, no comprehensive study has yet been carried out on the AP2/ERF gene family in rose (Rosa sp.), the most important ornamental crop worldwide., Results: The present study comprises a genome-wide analysis of the AP2/ERF family genes (RcERFs) in the rose, involving their identification, gene structure, phylogenetic relationship, chromosome localization, collinearity analysis, as well as their expression patterns. Throughout the phylogenetic analysis, a total of 131 AP2/ERF genes in the rose genome were divided into 5 subgroups. The RcERFs are distributed over all the seven chromosomes of the rose, and genome duplication may have played a key role in their duplication. Furthermore, Ka/Ks analysis indicated that the duplicated RcERF genes often undergo purification selection with limited functional differentiation. Gene expression analysis revealed that 23 RcERFs were induced by infection of the necrotrophic fungal pathogen Botrytis cinerea. Presumably, these RcERFs are candidate genes which can react to the rose's resistance against Botrytis cinerea infection. By using virus-induced gene silencing, we confirmed that RcERF099 is an important regulator involved in the B.cinerea resistance in the rose petal., Conclusion: Overall, our results conclude the necessity for further study of the AP2/ERF gene family in rose, and promote their potential application in improving the rose when subjected to biological stress.

Key Message: Rdr3 is a novel resistance gene of black spot in roses that maps to a chromosome 6 homolog. A new DNA test was developed and can be used to pyramid black spot resistance in roses. Diplocarpon rosae, the cause of rose black spot, is one of the most devastating foliar pathogens of cultivated roses (Rosa spp.). The primary method of disease control is fungicides, and they are viewed unfavorably by home gardeners due to potential environmental and health impacts. Planting rose cultivars with genetic resistance to black spot can reduce many of the fungicide applications needed in an integrated pest management system. To date, four resistance genes have been identified in roses (Rdr1, Rdr2, Rdr3, and Rdr4). Rdr3 was never mapped and is thought to be unique from Rdr1 and Rdr2. It is unknown whether it is an allele of Rdr4. To assess the novelty of Rdr3, a mapping population was created by crossing the Rdr3 containing cultivar George Vancouver with the susceptible cultivar Morden Blush. The mapping population was genotyped with the WagRhSNP 68 K Axiom array and mapped using the 'polymapR' package. Rdr3 was mapped to a chromosome 6 homolog confirming it is different from Rdr1 and Rdr2, found on chromosome 1, and from Rdr4, found on chromosome 5. The mapping information was used in conjunction with the Rosa chinensis genome assembly to develop new tightly linked SSRs for marker-assisted breeding. Three markers were able to predict the presence of Rdr3 in a 63-cultivar validation set. Additionally, 12 cultivars appear to have resistance genes other than Rdr3. The improved diagnostic markers will be a great asset to the rose-breeding community toward developing new black spot-resistant cultivars.

Background: The WRKYs are a major family of plant transcription factors that play roles in the responses to biotic and abiotic stresses; however, a comprehensive study of the WRKY family in roses (Rosa sp.) has not previously been performed., Results: In the present study, we performed a genome-wide analysis of the WRKY genes in the rose (Rosa chinensis), including their phylogenetic relationships, gene structure, chromosomal locations, and collinearity. Using a phylogenetic analysis, we divided the 56 RcWRKY genes into three subgroups. The RcWRKYs were unevenly distributed across all seven rose chromosomes, and a study of their collinearity suggested that genome duplication may have played a major role in RcWRKY gene duplication. A Ka/Ks analysis indicated that they mainly underwent purifying selection. Botrytis cinerea infection induced the expression of 19 RcWRKYs, most of which had undergone gene duplication during evolution. These RcWRKYs may regulate rose resistance against B. cinerea. Based on our phylogenetic and expression analyses, RcWRKY41 was identified as a candidate regulatory gene in the response to B. cinerea infection, which was confirmed using virus-induced gene silencing., Conclusions: This study provides useful information to facilitate the further study of the function of the rose WRKY gene family.

In the present study, we investigated the antifungal effects of engineered nanomaterials (ENMs) against Podosphaera pannosa (P. pannosa), a fungal pathogen that causes powdery mildew on plants in the rose family. Four commercial ENMs, including multi-wall carbon nanotubes (MWCNTs), reduced graphene oxide (rGO), copper oxide (CuO) nanoparticles (NPs) and titanium dioxide (TiO 2 ) NPs, were used to prepare 50 or 200 mg/L NP suspensions in deionized water. Rose leaves in water-agar plates were sprayed by different ENM suspensions mixed with P. pannosa conidia. After a 19-day standard infection test, the growth of P. pannosa on rose leaves was evaluated. All four ENMs inhibited infection by P. pannosa at the concentration 200 mg/L, whereas only CuO NPs decreased fungal growth at 50 mg/L. The phytohormone content of the leaves was measured across all treatments to investigate potential ENMs antifungal mechanisms. The results suggest that ENMs increased plant resistance to fungal infection by altering the content of endogenous hormones, particularly zeatin riboside (ZR). Our study demonstrates that ENMs exhibited distinctly antifungal effects against P. pannosa on roses, and could be utilized as a novel plant protection strategy after a comprehensive assessment of potential environmental risk., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Rose black spot is one of the most severe diseases of field-grown roses. Though R-genes have been characterised, little information is known about the molecular details of the interaction between pathogen and host. Based on the recently published genome sequence of the black spot fungus, we analysed gene models with various bioinformatic tools utilising the expression data of infected host tissues, which led to the prediction of 827 secreted proteins. A significant proportion of the predicted secretome comprises enzymes for the degradation of cell wall components, several of which were highly expressed during the first infection stages. As the secretome comprises major factors determining the ability of the fungus to colonise its host, we focused our further analyses on predicted effector candidates. In total, 52 sequences of 251 effector candidates matched several bioinformatic criteria of effectors, contained a Y/F/WxC motif, and did not match annotated proteins from other fungi. Additional sequences were identified based on their high expression levels during the penetration/haustorium formation phase and/or by matching known effectors from other fungi. Several host genotypes that are resistant to the sequenced isolate but differ in the R-genes responsible for this resistance are available. The combination of these genotypes with functional studies of the identified candidate effectors will allow the mechanisms of the rose black spot interaction to be dissected., (Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

During the last two years, greenhouse cultivation of rose (Rosa spp.) in the Netherlands has been challenged by an uncommon bacterial disease. Affected plants suffered from chlorosis, stunting, wilting, and necrosis. The bacterial isolates obtained from the different Rosa spp. cultivars were all identified as phylotype I, sequevar 33 from the 'Ralstonia solanacearum species complex' (RSSC), actually reclassified as Ralstonia pseudosolanacearum. The work in this paper considers the genetic diversity and the phylogenetic position of 129 R. pseudosolanacearum isolates from the outbreak. This was assessed by AFLP based on four different primer combinations and MLP using partial sequences of the egl, mutS, and fliC genes. The AFLP revealed identical profiles for all the isolates, irrespective of their association with Rosa sp. propagating material, Rosa spp. plants for cut flowers, or water used in the different greenhouse cultivations. These AFLP profiles were unique and diverged from profiles of all other reference isolates in the RSSC included. Furthermore, MLP on egl, fliC, and mutS gene sequences clearly demonstrated that all R. pseudosolanacearum isolates clustered in phylotype I, as a distinct monophyletic group. Interestingly, this monophyletic group also included phylotype I strain Rs-09-161 from eggplant (Solanum melongena), isolated in 2009 in India. AFLP and MLP were both efficient in revealing the genetic divergence from the RSSC isolates included. The phylogenetic tree constructed from the AFLP profiles was, in general, in agreement with the one obtained from MLP. Both phylogenetic trees displayed a similar clustering, supported by high posterior probabilities. Both methodologies clearly demonstrated that the R. pseudosolanacearum isolates from Rosa spp. grouped in a monophyletic group inside phylotype I, with a particular correspondence to a strain present in India, as revealed in MLP.

Peronospora sparsa is a downy mildew-causing oomycete that can infect roses, blackberries, and other members of the rose family. During the last 70 years, this disease has become a serious problem for rose growers in the U.S. and worldwide. While much is known about the disease and its treatment, including significant research on molecular identification methods, as well as environmental conditions conducive to disease and the fungicides used to prevent it, significant knowledge gaps remain in our basic comprehension of the pathogen's biology. For example, the degree of genetic relatedness of pathogen isolates collected from rose, caneberries, and cherry laurel has never been examined, and the natural movement of genotypes from host to host is not known. Further work could be done to determine the differences in pathogen population structure over time (using herbarium specimens and fresh collections) or differences in pathogen population structure and pathogen environmental adaptation for specimens from different geographic regions. The oospore stage of the organism is poorly understood, both as to how it forms and whether it serves as an overwintering structure in nurseries and landscapes. In production greenhouses, the detection of the pathogen using infrared thermographic imaging and possible inhibition by ultraviolet light needs to be explored. Further work needs to be done on breeding using wild roses as new sources for resistance and using new methods such as marker assisted selection and RNAi technologies. As roses are one of the most economically important ornamental crops worldwide, a proper understanding of the disease cycle could allow for better use of cultural and chemical controls to manage rose downy mildew in landscapes and in greenhouse and nursery production areas.

Black spot disease, which is caused by the ascomycete Diplocarpon rosae, is the most severe disease in field-grown roses in temperate regions and has been distributed worldwide, probably together with commercial cultivars. Here, we present data indicating that muRdr1A is the active Rdr1 gene, a single-dominant TIR-NBS-LRR (Toll/interleukin-1 receptor-nucleotide binding site-leucine rich repeat) (TNL)-type resistance gene against black spot disease, which acts against a broad range of pathogenic isolates independent of the genetic background of the host genotype. Molecular analyses revealed that, compared with the original donor genotype, the multiple integrations that are found in the primary transgenic clone segregate into different integration patterns in its sexual progeny and do not show any sign of overexpression. Rdr1 provides resistance to 13 different single-spore isolates belonging to six different races and broad field mixtures of conidia; thus far, Rdr1 is only overcome by two races. The expression of muRdr1A, the active Rdr1 gene, leads to interaction patterns that are identical in the transgenic clones and the non-transgenic original donor genotype. This finding indicates that the interacting avirulence (Avr) factor on the pathogen side must be widespread among the pathogen populations and may have a central function in the rose-black spot interaction. Therefore, the Rdr1 gene, pyramided with only a few other R genes by sexual crosses, might be useful for breeding roses that are resistant to black spot because the spread of new pathogenic races of the fungus appears to be slow., (© 2017 BSPP AND JOHN WILEY & SONS LTD.)

Application of 3.6 mm silicon (Si+) to the rose (Rosa hybrida) cultivar Smart increased the concentration of antimicrobial phenolic acids and flavonoids in response to infection by rose powdery mildew (Podosphaera pannosa). Simultaneously, the expression of genes coding for key enzymes in the phenylpropanoid pathway (phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, and chalcone synthase) was up-regulated. The increase in phenolic compounds correlated with a 46% reduction in disease severity compared with inoculated leaves without Si application (Si-). Furthermore, Si application without pathogen inoculation induced gene expression and primed the accumulation of several phenolics compared with the uninoculated Si- control. Chlorogenic acid was the phenolic acid detected in the highest concentration, with an increase of more than 80% in Si+ inoculated compared with Si- uninoculated plants. Among the quantified flavonoids, rutin and quercitrin were detected in the highest concentrations, and the rutin concentration increased more than 20-fold in Si+ inoculated compared with Si- uninoculated plants. Both rutin and chlorogenic acid had antimicrobial effects on P. pannosa, evidenced by reduced conidial germination and appressorium formation of the pathogen, both after spray application and infiltration into leaves. The application of rutin and chlorogenic acid reduced powdery mildew severity by 40% to 50%, and observation of an effect after leaf infiltration indicated that these two phenolics can be transported to the epidermal surface. In conclusion, we provide evidence that Si plays an active role in disease reduction in rose by inducing the production of antifungal phenolic metabolites as a response to powdery mildew infection.

Black spot disease of rose, incited by the fungus Diplocarpon rosae, is found worldwide and is the most important disease of garden roses. A gene-for-gene interaction in this pathosystem is evidenced by the presence of pathogenic races of D. rosae and the previous discovery of a dominant resistance allele at the Rdr1 locus in the diploid Rosa multiflora. The objective of the present study was to genetically analyze resistances to North American black spot races 3, 8, and 9 previously reported in tetraploid roses. Resistance to North American races 3 and 8 segregated 1:1 in multiple F(1) populations, indicating that both are conferred by dominant alleles at single loci and are present in simplex (Rrrr) configuration. Gene pyramiding was demonstrated by combining both resistances into single genotypes. Resistance to race 9 was partial and segregated in a quantitative fashion. Analysis of these populations with microsatellite markers previously developed for Rdr1 revealed that the gene conferring race 3 resistance resides within the same R gene cluster as Rdr1. Race 8 resistance segregated independently and is, therefore, a novel locus for black spot resistance in rose which we have named Rdr3. NBS and LRR profiling were used in a bulked segregant analysis to identify a marker 9.1 cM from Rdr3, which was converted to a SCAR marker form for marker-assisted breeding.

A strategy combining dual-suppression PCR and thermal asymmetric interlaced PCR was used to determine sequences flanking microsatellite regions in Pythium helicoides. The primer pairs were designed to amplify loci containing (AC)n, (GA)n, (AGC)n, (CAC)n(CAA)n, (TCA)n and (CTTT)n repeats from the P. helicoides nuclear genome. The PCR products of each primer pair, amplified from three representative isolates collected from different hosts and locations, were cloned and sequenced. Different degrees of polymorphism were detected among these microsatellite markers. The numbers of alleles were 6, 2, 4, 11, 4 and 4 in YL-AC, YL-AGC, YL-CAA, YL-CTTT, YL-GA and YL-TCA, respectively. Allele analysis of 30 P. helicoides isolates showed length polymorphisms in all loci, except for YL-AC, using capillary electrophoresis. Thus, we have developed a simple method for designing PCR primers to amplify microsatellite markers from P. helicoides.

Powdery mildew (Podosphaera pannoso) is one of the most serious fungal diseases on both greenhouse and field grown roses. Improvement of disease resistance is a major selection aim for garden rose breeders. For rose cultivars, being mostly tetraptoid, it is complicated to develop molecular markers for resistance. Hence, a segregating diploid population was established from a cross between 'Yesterday', a commercial available rose variety susceptible to powdery mildew, and R. wichurana, a rose species with resistance to certain isolates of powdery mildew. A progeny of 94 seedlings was planted in the field. The segregation of powdery mildew resistance was studied in this population by means of a bioassay with two different monoconidial isolates of powdery mildew. Based on the response to these inoculations different groups were selected: a first group of genotypes was susceptible to both isolates, other groups were susceptible to one of both isolates and a last group was resistant to both tested isolates. The disease resistance inherits for both isolates in a quantitative way. A genetic map based on AFLP and SSR markers was established and will be used for QTL analysis of powdery mildew resistance.

Podosphaera pannosa, the causal agent of rose powdery mildew, hampers the production of cut roses throughout the world. A major tool to control this disease is the use of resistant plant material. Single resistance genes, like Rpp1, may be overcome within a few years by high risk pathogens like powdery mildews. Durable resistance could be achieved using quantitative resistances. Here we describe mapping of QTLs for resistance to P. pannosa in six different environments (artificial and natural infections in the greenhouse over 3 years and natural infections in the field over 2 years). AFLPs, RGAs and other marker types were used to construct an integrated linkage map for the diploid population 97/7 containing 233 markers. In a selective genotyping procedure, marker segregation was analysed for 170 of the up to 270 phenotyped individuals. We identified seven linkage groups with an average length of 60 cM, corresponding to seven rose chromosomes in the haploid set. Using an LOD significance threshold of 3.9 we detected a total of 28 QTLs for the nine powdery mildew disease scores under analysis. Using the data from artificial inoculations with powdery mildew race 9, three resistance QTLs explaining about 84% of the variability were mapped. Twelve and 15 QTLs were detected for resistance to naturally occurring infections in the greenhouse and in the field, respectively, over several years.

Among downy mildews occurring on ornamentals in Poland the most dangerous are downy mildew of rose and downy mildew of German statice (Tartarian statice). Downy mildew of rose caused by Peronospora sparsa Berk. is a serious threat to commercial cultivation of cut roses, especially grown under plastic tunnels. Peronospora statices Lobik casual agent of German statice downy mildew can causes the total losses in the second year of statice cultivation more than 70%. Both pathogens are very difficult to control. Effectiveness of azoxystrobine, cymoxanil + famoxate, mancozeb, phosethyl aluminium, phosethyl aluminium + fenamidone, propamocarb in the control of P. sparsa and P. statices was presented. In the control of statice downy mildew none of tested compounds was able to control satisfactory the pathogen. Relatively the best results were obtained with mixture of fenamidone (88 microg/cm3) and phosethyl AL (1334 microg/cm3). In the control of rose downy mildew, the best results were obtained with phosethyl Al at concentration 1600 microg/cm3.

Rust fungi in the genus Phragmidium are frequent pathogens of both wild and cultivated roses. We investigated the occurrence and relationships of rusts on dog roses, Rosa sect. Caninae (Rosa canina, R. corymbifera and R. rubiginosa) in Germany. Two Phragmidium species, P. mucronatum and P. tuberculatum, were able to infect each of the three dog rose species examined. However, the overall infection of R. rubiginosa was significantly lower, which could be important for rose breeding. Despite overlapping host ranges, the evolutionary background of P. tuberculatum and P. mucronatum is quite distinct. Phylogenetic analyses of the D1/D2 region of the LSU rDNA suggest that P. mucronatum shares a common ancestor with other rose rusts, whereas P. tuberculatum evolved from a Rubus-Sanguisorba rust clade and must have undergone a host shift to Rosa spp.

In the control of Sphaerotheca pannosa var. rosae on rose Tolylfluanide + tebuconazole (Folicur Multi 50 WG at concentration 0.1%) was used for spray 2 -times at 14-day-intervals or 4-times at 7-day-intervals. After 4 weeks of plants protection effectiveness of tested product was about 75%. In the control of Diplocarpon rosae, the product was applied when first disease symptoms appeared on rose shrubs. Application was repeated 5-times at 14-day-intervals or 9-times at 7-day-intervals. After 9 week-protection effectiveness of tested product was about 85% and depended on frequency of sprayings. In the control of Puccinia horiana on chrysanthemum, Tolylfluanide + tebuconazole was used as plant spray twice at 14-day-intervals or 4-times at 7-day-intervals. After 4 week-protection the product suppressed of new telia formation about 55%. Application of the product for willow rust (Melampsora epitea) control suppressed formation of new uredia about 86% and half of them were dried. In the control of pelargonium rust (Puccinia pelargonii-zonalis) the product was used as plant spray 4-times at 7-day-intervals. It suppressed formation of new uredia about 90% and 1/3 of them were dried. It was found that 1 or 7 days after rose spray, spores of D. rosae collected from leaf blades only in 6% germinated. Spores taken from nonspraying leaves germinated in 90%. In case of P. pelargonii-zonalis, after 1 or 7 days after spraying, spores collected from protected plants germinated at 3%, compared to 90% on untreated plants. Spores of B. cinerea, collected from protected plants germinated at about 10%, whereas on control leaves at 90%.

Chitosan, (Biochikol 020 PC) a potential elicitor of plant defence and also an active inhibitor of fungal growth was used in experiments. The compound was used at concentrations 0.01 to 0.2% as a plant spray for rose protection against Sphaerotheca pannosa var. rosae, Peronospora sparsa and Diplocarpon rosae, Oidium chrysanthemi and Puccinia horiana on chrysanthemum, against Melampsora epitea on willow. Myrothecium roridum on dieffenbachia and against Lophodermium spp on Scots pine. Effectiveness of this product, applied curatively 2-times at 7-day-intervals after appearance of rose powdery mildew symptoms on most of plant parts, was about 32%. When compound was applied just after the first disease symptoms appearance, after 2-week-protection its effectiveness was about 43-60%. After 4 weeks, effectiveness of chitosan (Biochikol 020 PC) increased and ranged from 67 to 77%. In the control of P. sparsa on rose shrubs, growing in plastic tunnel, chitosan at conc. 0.025% was applied 4 times at weekly intervals. Effectiveness of chitosan against this pathogen was over 72%, similar as standard fungicide. Increase of chitosan (Biochikol 020 PC) concentration resulted in decrease of its effectiveness. In case of D. rosae control, chitosan used 9 times at weekly intervals was applied after first disease symptoms. After 3-week-protection effectiveness of the compound varied from 18 to 60% according to used concentration. After 9 weeks of protection effectiveness of tested product ranged from 16 to 23% and was connected with used concentration. Increase of chitosan concentration resulted in better protection of rose shrubs. Effectiveness of chitosan (Biochikol 020 PC) used 4-times at conc. 0.01 to 0.05% as chrysanthemum spray in the control of Oidium chrysanthemi ranged from 69 to 79%, whereas against Puccinia horiana from 54-97%. Two-time-spraying of willow with rust symptoms (Melampsora epitea) with chitosan at conc. 01-0.04% caused decrease of disease severity from 26 to 47%. Additionally about 10 to 25% of uredinia were dried up. Chitosan (Biochikol 020 PC) applied for leaf spraying inhibited the development of Myrothecium leaf spot on dieffenbachia more than 85%. Curative application of the compound controlled Myrotecium leaf spot at about 40%. Chitosan efficacy was also tested against one-year-old Pinus sylvestris seedlings growing in field conditions. Plants were sprayed fortnightly 14-times (long programme) or 5-times (short programme) against Lophodermium spp. Observations made during next spring showed that effectiveness of chitosan (Biochikol 020 PC) was more than 50% (long programme) and below 50% (short programme).

An antimicrobial protein gene, Ace-AMP1, was introduced into Rosa hybrida cv. Carefree Beauty via Agrobacterium-mediated transformation. A total of 500 putative transgenic plants were obtained from 100 primary embryogenic calli co-cultivated with A. tumefaciens following selection on a regeneration medium containing 100 mg/l kanamycin. Polymerase chain reaction analysis of these putative transgenic lines, using primers for both Ace-AMP1 and neomycin phosphotransferase ( npt II) genes, showed that 62% of these plants were positive for both transgenes. These lines were further confirmed for stable integration of Ace-AMP1 and npt II genes by Southern blotting. Transcription of the Ace-AMP1 transgene in various transgenic rose lines was determined using Northern blotting. Transgenic rose lines inoculated with conidial spores of Sphaerotheca pannosa (Wallr.: Fr.) Lev. var. rosae showed enhanced resistance to powdery mildew using both a detached-leaf assay and an in vivo greenhouse whole-plant assay.

Powdery mildew, caused by Podosphaera pannosa, is one of the most-severe diseases of roses grown under glass. The differentiation into physiological races and the genetic analysis of resistance in a segregating host population was investigated using single conidial isolates of the pathogen. Using ten rose genotypes, all eight isolates of the pathogen could be ascribed to different races. Five races were isolated from one location, which indicates that populations of P. pannosa exhibit a high racial diversity. Infection experiments in a backcross-population of 114 rose plants resulted in a 1:1 segregation, suggesting control by a single dominant gene. Rpp1 is the first resistance gene against rose powdery mildew to be described.

The constant input of chemical fungicides against foliar diseases in glasshouse crops has instigated research on alternative control substances. Various salts have been mentioned in the literature and we have tested several bicarbonates, sulphates and phosphates against powdery mildew fungi in cucumber, tomato, sweet pepper and rose and against Bremia lactucae in lettuce. The experiments were run according to EPPO guidelines for efficacy in glasshouses of Applied Plant Research (PPO). Plants were sprayed weekly and disease and yield were assessed. In cucumber, NaHCO3, KHCO3, MgSO4 and MnSO4 all reduced powdery mildew significantly, with MnSO4 as the most effective agent. In tomato, MgSO4, MnSO4 and KH2PO4 all reduced powdery mildew by approx. 40% whereas NaHCO3 and KHCO3 gave approx. 95% control. In pepper, NaHCO3 controlled Leveillula taurica by 50-90% compared to the untreated control. In rose, the efficacy of KH2PO4, MnSO4, MgSO4, NaHCO3 and KHCO3 against powdery mildew differed between cultivars. In general, the bicarbonates were the most effective compounds. In lettuce, NaHCO3 reduced Bremia severity by 70 to 80%. The conclusion of this range of experiments is that especially bicarbonates can be a good alternative for control of foliar fungal diseases in glasshouse crops. The effective concentration is not causing any symptoms of phytotoxicity, provided that spraying is not done at high irradiation conditions. Bicarbonates have been submitted for the Dutch list of exceptions on the Pesticide Law.

Production of disease-free propagation material is a major means of controlling most bacterial diseases of plants, particularly when neither resistant clones nor effective chemical treatments are available. For this purpose sensitive, specific and rapid detection methods are required. The advent of molecular biology and, in particular, the polymerase chain reaction (PCR) has opened new ways for the characterization and identification of plant pathogens and the development of disease-management strategies. PCR-based detection methods rely on the development of primers for the specific detection of the pathogen. The use of pathogenicity genes as targets for primer design is the preferred procedure for obtaining specific primers but other procedures may also be useful for this purpose. In the present review we describe four examples of procedures for detecting four important bacterial pathogens in Israel: Erwinia herbicola pv gypsophilae in gypsophila, Xanthomonas campestris pv pelargonii in geranium, Agrobacterium tumefaciens in asters and roses, and Xanthomonas campestris pv campestris in crucifers. Procedures for constructing specific PCR primers for each bacterium are illustrated and discussed as well as the combination of PCR with other methods.