Carbendazim resistance in field isolates of Sclerotinia sclerotiorum in China and its management

The ascomycete fungus Sclerotinia sclerotiorum is a devastating necrotrophic plant pathogen with an unusually broad host range and worldwide distribution. The benzimidazole fungicide carbendazim has been widely used to control S. sclerotiorum in China for more than three decades and high levels of carbendazim resistance have been reported in eastern China. In this study, carbendazim sensitivity was assessed in a total of 5042 field isolates of S. sclerotiorum collected from different geographical regions of China from 2008 to 2013. Results showed that no carbendazim resistant isolates could be detected in northwestern and northeastern China. In 2013, 0.77% and 0.72% of the field isolates assayed were highly resistant to carbendazim in Hunan and Hubei provinces of central China, respectively. In Anhui province of eastern China, the frequencies of carbendazim resistance were 18.18%, 7.05% and 7.25% in 2008, 2011 and 2012, respectively. The decrease in frequency of carbendazim resistance in Anhui province from 2008 to 2012 was statistically significant (P = 0.001). Nevertheless, no significant differences (P ≥ 0.16) were found in fitness parameters such as mycelial growth on PDA media, virulence to oilseed rape plants and oxalic acid production between carbendazim resistant and sensitive isolates. A negative cross resistance pattern was detected between carbendazim and diethofencarb. Mixture of carbendazim and diethofencarb (4:1) applied at 200 μg/mL provided 100% and 91.5% preventive efficacy against carbendazim resistant and sensitive isolates, respectively, and 87.1% and 81.7% curative efficacy against resistant and sensitive isolates, respectively.     

Sensitivity of Cercospora beticola isolates from Serbia to carbendazim and flutriafol

Cercospora beticola, causal agent of Cercospora leaf spot (CLS) of sugar beet, is primarily controlled by fungicides. Benzimidazole and demethylation inhibiting fungicides, including carbendazim and flutriafol, have been widely used in Serbia. Since these fungicide groups have a site-specific mode of action, there is a high risk for developing resistance in target organisms, which is the most important limiting factor in Cercospora leaf spot chemical control. A rapid identification of flutriafol and carbendazim resistance can help researchers in examining the potential of different fungicide resistance management practices, as well as in selection of fungicides for use in the areas where resistance has occurred. One hundred single-conidia isolates were collected from 70 representative locations of the sugar beet production region in Serbia. Evaluation of the isolates' sensitivity was based on the reduction of mycelial growth on medium amended with 1.25 μg mL⁻¹ flutriafol and 5 μg mL⁻¹ carbendazim. Resistance to flutriafol and carbendazim was detected in 16% and 96% of the tested isolates, respectively. All isolates resistant to flutriafol were resistant to carbendazim as well, which is the first report of a double resistance to fungicides in C. beticola. Detection of the isolates resistant to flutriafol and carbendazim using Cleaved Amplified Polymorphic Sequence (CAPS) markers confirmed the results of the in vitro tests. The efficacy of carbendazim, flutriafol, azoxystrobin, and tetraconazole at commercially recommended doses was evaluated in field trials where sugar beet plants in plots were inoculated with a mixture of isolates either sensitive and/or resistant to flutriafol and carbendazim. Carbendazim and flutriafol efficacy was very low in plots inoculated with isolates resistant to these fungicides. Presented results will contribute to development of a pathogen population sensitivity monitoring strategy that could be used for an effective CLS management in the region.     

Multiple-fungicide resistance to carbendazim,diethofencarb, procymidone,and pyrimethanil in field isolates of Botrytis cinerea from tomato in Henan Province,China

Botrytis cinerea, the fungal pathogen responsible for causing gray mold in tomatoes, frequently exhibits multiple fungicide resistance. In this study, the sensitivity to carbendazim (Car), diethofencarb (Die), procymidone (Prc), and pyrimethanil (Pyr) was determined for 263 isolates of B. cinerea from diseased fruit and leaves of tomato plants from 87 commercial greenhouses in Henan Province of China during 2013 and 2014. Of the isolates tested, 89%, 93%, 85%, and 86% were resistant to Car, Die, Prc, and Pyr, respectively. Twelve phenotypes were found having different sensitivity patterns, and multi-fungicide resistance was found to one, two, three or four fungicides. The percentage of multiple fungicide resistant isolates (Car^RDie^RPrc^RPyr^R) was 68%. Mycelial growth, spore production, and mycelial dry weight were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. However, the virulence of Car^SDie^RPrc^SPyr^S isolates was higher than that of the other fungicide-resistance phenotypes. All of the carbendazim and diethofencarb resistant phenotypes carried a point mutation at codon 198 (E198A or E198K) or 200 (F200Y) in the β-tubulin gene, and all of the procymidone resistant phenotypes carried point mutations at codons 369 (Q369P) and 373 (N373S) in the Bos1 gene. These results provided important reference data for assessment of resistance risk of B. cinerea isolates from commercial tomato transplants in Henan Province, indicating the existence of a high frequency of resistance to carbendazim, diethofencarb, procymidone, and pyrimethanil revealing the necessity of a more integrated control of gray mold.     

Characterization of fungicide resistant isolates of Phaeoacremonium aleophilum infecting grapevines in Spain

Fungicide resistance in Phaeoacremonium aleophilum, one of the most frequent fungal pathogens associated with grapevine trunk diseases, was investigated and found to exist in some isolates of the pathogen against a commercial formulation, Escudo^®. The effect of this compound and its two active substances, carbendazim and flusilazole, was first evaluated on the mycelial growth of P. aleophilum. Escudo^®-resistant isolates were estimated at a frequency of 24% in Spanish vineyards. Then, the two active substances were used individually to test their effect on mycelia growth of twelve single-spore cultures originating from six Escudo^®-resistant isolates. Flusilazole (DMI-triazole) did not inhibit mycelia growth of any single-spore cultures of P. aleophilum. Carbendazim (benzimidazole) used alone allowed the growth of the same single-spore cultures that were also resistant to Escudo^®. AFLP characterization of sensitive and resistant single-spore cultures showed genetic diversity within P. aleophilum isolates but no AFLP markers were associated with resistance. New primers set (L2/R1) were designed to partially amplify the exon 6 of the beta-tubulin gene of P. aleophilum. Two different point mutations resulted in glycine (GGC) or lysine (AAA) replacing the glutamic acid (GAG) at codon 198 of the beta-tubulin gene in some of the resistant single-spore cultures studied. Resistant single-spore cultures of P. aleophilum were shown to have different aggressiveness levels as sensitive single-spore cultures by inoculation of wood segments of Vitis vinifera in the presence and absence of fungicide.     

Sensitivity to fluopicolide of wild type isolates and biological characteristics of fluopicolide-resistant mutants in Pseudoperonospora cubensis

Cucurbit downy mildew caused by the oomycete pathogen Pseudoperonospora cubensis is a devastating disease that is distributed worldwide and affects cucumber in open fields and greenhouses. Fluopicolide, which was a novel systemic fungicide and was released in 2008, it is very effective in controlling downy mildew on cucumber and grape, potato late blight and pepper Phythophthora blight and reduces the loss caused by the diseases, but so far the potential for P. cubensis to develop resistance to fluopicolide has not been investigated. Hence, a laboratory study was undertaken to assess the risk of P. cubensis developing resistance to fluopicolide. Baseline sensitivity to fluopicolide was determined by using 75 P. cubensis isolates collected from cucumber-growing greenhouses in Hebei province, where no fluopicolide had been used for control of cucumber downy mildew before. Values of effective concentrations for 50% inhibition (EC₅₀) of sporulation ranged from 0.02 to 0.40 μg ml⁻¹ and were distributed as a unimodal curve, indicating that all 75 isolates were sensitive to fluopicolide. Sporangia of nine sensitive isolates were ultraviolet (UV)-irradiated, and four fluopicolide-resistant mutants were acquired at a mutation frequency of 7.4 × 10⁻⁷. Seven mutants resistant to fluopicolide were obtained from seven isolates by sporangia adaptation on fluopicolide-treated leaves of cucumber. The EC₅₀ values for all eleven fluopicolide-resistant mutants ranged from 3.37 to 13.06 μg ml⁻¹ with mean resistance factors of 7.9–118.0. After 10 sporangia transfers on fungicide-free leaves of cucumber, all eleven resistant mutants remained resistant to fluopicolide with mean resistance factors of 8.2–81.3. Seven resistant mutants from the selection for resistance and one resistant mutant from UV mutagenesis exhibited stable resistance; however, the other three resistant mutants from UV irradiation became significantly less resistant. Compared to their respective sensitive parents, the eleven resistant mutants exhibited diversity in latent period, infection frequency, lesion extension and sporulation ability. Five out of the eleven resistant mutants exhibited prolonged latent period and three out of the eleven resistant mutants provided decreased infection frequency (IF) compared to their respective parents, indicating that in some cases, resistance mutation might affect the latent period and IF of P. cubensis. There were significant differences in pathogenicity and ability to produce sporangia, but this seemed not to be caused by resistance mutation. No cross-resistance was detected between fluopicolide and azoxystrobin, metalaxyl, dimethomorph, or cymoxanil. In all, there could be a moderate to high risk of field populations of P. cubensis developing resistance to fluopicolide, and populations of P. cubensis should be monitored regularly for their shift of sensitivity over years of application.     

Effect of leaf rust (Puccinia recondita tritici) on yield of four isogenic wheat lines

Four near-isogenic lines of wheat (Thatcher and Thatcher backcross lines with Lr3b, Lr16 and Lr9) with different levels of resistance to leaf rust were used to determine the relationship between disease severity and yield loss in the greenhouse and the field. Larger areas under the disease progress curves were associated with greater host susceptibility. Yield losses varied from 21% to 47% depending on the susceptibility of the lines and the severity of the epidemics. Yield loss measured varied with the size of the plot harvested. Percentage yield losses from each line in the field experiment were pooled and regressed against the area under the disease progress curve of disease severity on flag leaf and the mean severity per tiller. Both regressions were highly significant with R² of 0·89 and 0·88, respectively. The ratio of area under the disease progress curve to yield loss was higher in susceptible than resistant cultivars; thus it was difficult to ascertain whether a linear or S-shaped curve was more appropriate for the disease-loss relationship.     

Efficacy of Bacillus subtilis MBI 600 Against Sheath Blight Caused by Rhizoctonia solani and on Growth and Yield of Rice

Rice sheath blight disease(ShB),caused by Rhizoctonia solani,gives rise to significant grain yield losses.The present study evaluated the efficacy of Integral?,the commercial liquid formulation of Bacillus subtilis strain MBI 600,against rice ShB and for plant growth promotion.In greenhouse studies,four log concentrations of Integral(from 2.2×106 to 2.2×109 cfu/mL) were used as seed treatment(ST).After 25 d,seedlings were dipped(SD) into Integral prior to transplanting.At 30 d after transplanting(DAT),leaf sheaths were inoculated with immature sclerotia of the pathogen.At 45 DAT,a foliar spray(FS) with Integral was applied to some treatments.The fungicide control was 50% carbendazim at 1.0 g/L,and a nontreated control was also included.Overall,there were 10 treatments,each with five replications.ShB severity was rated at 52 DAT,and seedling height and number of tillers per plant were rated at 60 DAT.In 2009,two field trials evaluated Integral at 2.2×108 and 2.2×109 cfu/mL.Integral was applied as ST,and seedlings were produced in a nursery bed.After 32 d,seedlings were treated with Integral as SD and transplanted into 10 m2 blocks.Foliar sprays were given at 45 and 60 DAT.There were seven treatments,each with eight replications arranged as a factorial randomized complete block design.At 20 DAT,the plots were broadcast inoculated with R.solani produced on rice grains.Seedling height before transplanting,ShB severity at 90 DAT,and grain yield at harvest were recorded.Integral at 2.2×109 cfu/mL provided significant increase of seedling heights over other treatments under greenhouse conditions.The Integral treatments of ST + SD + FS at 2.2×109 cfu/mL significantly suppressed ShB over other treatments.In field studies,Integral provided significant increase of seedling height in nursery,and number of tillers per plant,compared with the control.ShB severity was significantly suppressed with higher concentrations of Integral compared to lower concentrations.Grain yield were the highest at an Integral concentration of 2.2×109 cfu/mL.Overall,Integral significantly reduced ShB severity,enhanced seedling growth,number of tillers per plant and grain yield as ST + SD + FS at the concentration of 2.2×109 cfu/mL under the conditions evaluated.     

Baseline sensitivity of Monilinia laxa from Greece to fenhexamid and analysis of fenhexamid-resistant mutants

Fenhexamid is a hydroxyanilide fungicide with excellent performance against Botrytis cinerea but also effective against Monilinia spp. which cause brown rot disease in apple and stone fruit. A total of 75 Monilinia laxa field isolates were utilized to determine baseline sensitivity while a number of fenhexamid-resistant laboratory mutants were used to evaluate the resistance risk associated with the longevity of the effectiveness of fenhexamid. Fenhexamid was found to be highly effective against all field isolates. EC₅₀ values ranged from 0.02 to 1 μg mL⁻¹ and were distributed unimodaly around an average of 0.1 μg mL⁻¹. M. laxa laboratory strains with moderate and high resistance levels to fenhexamid were isolated after UV mutagenesis. All fenhexamid-resistant strains showed parental sensitivity to carbendazim, iprodione, fludioxonil, pyraclostrobin, flusilazole and prochloraz. Interestingly, some of the mutant strains were also resistant to tridemorph, fenpropimorph and spiroxamine. Studies on fitness parameters of fenhexamid-resistant strains revealed a fitness cost on sporulation and pathogenicity but not on mycelial growth. These results suggest that fenhexamid should be a good alternative site-specific fungicide for the control of brown rot disease caused by M. laxa. However, appropriate anti-resistance strategies should be considered to ensure the successful commercial use of fenhexamid in the long run.     

An <Emphasis Type="Italic">In Vitro</Emphasis> Assay Method for Resistance to Bacterial Wilt (<Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>) in Potato

To develop an in vitro assay method for bacterial wilt resistance in potato, resistant and susceptible standard genotypes were grown in vitro, and different conditions of inoculation with Ralstonia solanacearum phylotype I/biovar 4 were examined. The optimal condition was the inoculation of 6–8 leaf stage plants with a bacterial concentration of 10² CFU ml^?1 and an incubation temperature of 28 °C. Evaluation of stem wilting was more reliable than that of leaf wilting. Using this method, nine genotypes with different resistance levels in the field were evaluated. Lower disease indices were obtained for genotypes with high resistance levels in the field, suggesting that this assay is useful for evaluating bacterial wilt resistance in a controlled environment.     

Yield traits of six clones of Miscanthus in the first 3 years following planting in Poland

Miscanthus species are highly productive with low inputs and are excellent candidates for bioenergy feedstock production. A field experiment was conducted to characterize phenotypic differences in selected clones generated from interspecific hybrids of Miscanthus sinensis × Miscanthus sacchariflorus and intraspecific hybrids within M. sinensis. The field experiment was planted in plots of 20 m² at a density of 1 plant m⁻² in three randomized blocks. The trial was monitored for 3 years for traits important to biomass production including plant height, tiller density, tuft diameter and shoot diameter. ANOVA showed significant genotypic variation in these traits once the stand was 2 years old. This study shows that tillering and tuft diameter in years 1 and 2 are the most important traits influencing biomass yield, but over 3 years when the highest yielding potential is reached, tillering and tuft diameter have the highest correlation with biomass yield. These results identifying high-yielding Miscanthus clones will be utilized in our plant improvement program.     

Larval survival and plant injury of Cry1F-susceptible, -resistant,and -heterozygous fall armyworm (Lepidoptera: Noctuidae) on non-Bt and Bt corn containing single or pyramided genes

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target of transgenic corn, Zea mays L., expressing Bacillus thuringiensis (Bt) proteins in both North and South America. A highly Cry1F-resistant strain of S. frugiperda was established from a field collection in Puerto Rico in 2011. In this study, three greenhouse trials were conducted to evaluate larval survival and leaf injury of Cry1F-susceptible, -resistant, and -heterozygous genotypes of S. frugiperda on whole plants of five non-Bt and eight Bt corn hybrids. The Bt corn products included two single-gene Bt corn hybrids containing Herculex^®I (Cry1F) and YieldGard^® (Cry1Ab) traits and six pyramided Bt corn hybrids representing four traits: Genuity^® VT Double Pro™, Genuity^®VT Triple Pro™, Genuity^® SmartStax™, and Agrisure^® Viptera™ 3111. In each trial, neonates of S. frugiperda were placed into the plant whorls at vegetative plant stages (V6–V10). Larvae of the three insect genotypes on non-Bt corn hybrids survived well and caused serious plant injury. Cry1Ab corn was ineffective against all three insect genotypes. On Cry1F corn plants, resistant larvae survived on 72.9% plants after 12–15 d and caused a leaf injury rating (Davis' 1 to 9 scales) of 5.7 after 7 d and 7.6 after 12–15 d. Both the larval survivorship and leaf injury rates of the resistant larvae on Cry1F corn plants were not significantly different from those observed on non-Bt corn hybrids. In contrast, no live larvae and little or no leaf injury were observed on the Cry1F corn plants that were infested with susceptible or heterozygous genotypes, or on the pyramided Bt plants. The results demonstrated that the Cry1F-resistant S. frugiperda was highly resistant to whole plants of Cry1F corn and the resistance was recessive. Hybrids that contained one of the four pyramided Bt traits were effective for managing the Cry1F resistance in S. frugiperda.     

New host resistances to Pseudocercosporella capsellae and implications for white leaf spot management in Brassicaceae crops

Effective host resistance is the most cost-effective long term prospect for successful management of white leaf spot disease (Pseudocercosporella capsellae) in Brassicaceae. In two separate field trials, 168 genotypes were screened. In the first trial, lines of Brassica oleracea var. capitata (59), Brassica napus (34), Brassica juncea (6) and B. juncea containing wild weedy Brassicaceae introgression(s) (14) were arranged; and in the second, Australian historic and current B. napus (45) and B. juncea (10) varieties were screened. There was wide variation in expression of resistance, from complete resistance to highly susceptible as assessed by two disease parameters, viz. (i), Area Under Disease Progress Curve (AUDPC) for percent leaves diseased (values 0–221.2) and (ii) Percent Leaf Collapse Index (%LCI) values for leaf collapse due to disease (0–38.7). Brassica oleracea var. capitata was overall the most resistant species, while B. juncea the most susceptible with the majority having AUDPC values >75 and B. napus was intermediate. Five B. oleracea var. capitata genotypes were completely resistant, with 0 AUDPC and %LCI values. Pioneer^® 45Y22 (RR) ‘Mystic’ and ‘Wahoo’ were also highly resistant, with the least %LCI (<3.7) and AUDPC (<20) of the Australian B. napus varieties. In contrast, ‘Thunder TT’ (AUDPC -133.6; %CLI – 15.6) and ‘Carbine’ (AUDPC – 73.8; %CLI – 12.5) were the most susceptible lines in first and second trials, respectively. The particularly high susceptibility of newly released B. juncea varieties such as ‘Xceed OasisCL’ highlights the risk of significant losses in such susceptible varieties when deployed in areas with high degree of pressure for white leaf spot disease. There was no association between AUDPC or % CLI with year of Australian varietal release, indicating that Australian breeding programs not made improvement for resistance to white leaf spot over the past two or more decades. Resistant varieties identified in this experiment can now not only be utilized in breeding programs to significantly improve overall crop resistance and management of white leaf spot disease, but also directly deployed to lower the severe inoculum load challenging current varieties.     

Resistance of Botrytis cinerea Pers. and Plasmopara viticola (Berk. & Curt.) Berl. and de Toni to fungicides in French vineyards

The control of Botrytis cinerea and Plasmopara viticola, two of the most important parasites on grapevines, is achieved by using fungicides. Recently several highly active compounds have been introduced but with some of them, failures of disease control have been observed in French vineyards. These phenomena are due to the build-up of resistance to benzimidazoles (benomyl, carbendazim, thiophanate-methyl) or to di-carboximides (iprodione, procymidone, vinclozolin) in Botrytis cinerea and to phenylamides (metalaxyl, ofurace) in Plasmopara viticola. The characteristics of the various resistant strains (levels of resistance, patterns of cross-resistance, fitness) are described. In 1982 and 1983 there has been monitoring for resistance in most French vineyards: the tests used and the results with regard to the frequencies of resistant strains are discussed.     

Inheritance of early blight resistance from open-pollinated 4×-2× potato hybrids

Early blight, caused byAlternaria solani Sorauer, is a serious disease of potatoes that occurs in most potatogrowing regions in the world. There is little resistance to early blight among commercial potato cultivars. However, resistance to early blight in diploid (Solanum phureja-S. stenotomum) potatoes has been identified, and was found to be highly heritable and readily transferred to the tetraploid level via4×-2× crosses. The purposes of this study were to identify good levels of early blight resistance in open-pollinated4×-2× (Solanum tuberosum×S. phureja-S. stenotomum) hybrids selected for horticultural characteristics, to estimate broad-sense heritability for early blight resistance in these hybrids, and to investigate the general and specific combining ability for resistance to early blight from some of these early blight resistant tetraploid selections. Four early blight resistant clones were crossed as female parents with four different male parents in a design II mating scheme to generate 16 families. Approximately 20 randomly chosen offspring per family were visually evaluated for early blight resistance in 1995 and 1996 in a randomized complete block design in Pennsylvania. Broad-sense heritability for early blight resistance among the open-pollinated4×-2× hybrids which were originally selected for horticultural characteristics was 0.91 with a 95% confidence interval of 0.80, 0.94. There were significant differences among female and male parents for area under the disease progress curve, indicating the importance of general combining ability for early blight resistance in this germplasm. The female x male source of variation was not significant, indicating that specific combining ability was not important. The greatest number of resistant progeny were observed in families where both parents were derived from the early blight resistant population; however, at least one highly resistant progeny was produced in all families. These results suggest that the early blight resistance in these clones can be readily incorporated into the commercial tetraploid breeding population.     

A simple and efficient method to assess susceptibility of potato to stem rot byErwinia carotovora subspecies

Limited development of blackleg resistant potato cultivars has been ascribed to the lack of an effective and reliable test for large scale blackleg resistance screening. A method using leaf petioles was developed for assessing susceptibility of potato stems to blackleg pathogens (Erwinia carotovora subspeciesatroseptica andE. c. subspeciescarotovora). Fifty grams of sterile autoclaved silica sand in Magenta jars (GA-7 vessel, Magenta Corp., Chicago, Illinois) was drenched with bacterial suspension, (17 ml, 2.6 × 10 to 2.6 × 10⁷ cfu/ml) and freshly cut petioles from 5 to 6 week-old plants were inserted to about 5 mm into the sand. The Magenta jars with lids closed were kept on a laboratory bench (20 ±2 C, 16 hours light). The arrangement provided near ideal post-inoculation incubation environment of high humidity for pathogen multiplication and rot development in the petioles. Length of rot was recorded 72 hours after inoculation. Linear regression of rot development on inoculum concentration had R² ≥ 0.80. The petioles inoculated and incubated by this method produced measurable rot lesions with a bacterial concentration as low as 2.6 × 10⁴ cfu/ml and differentiated between the resistant cultivar Russet Burbank and the susceptible cultivar Sangre at all the inoculum concentrations used. Furthermore, susceptibility ranking of five cultivars obtained by this method agreed in general with their reported field reactions. The method is simple and sensitive, and could be used for large scale screening for stem rot resistance in wildSolanum species and early generation breeding populations.     

Effects of fungicides used to control Rhynchosporium secalis where benzimidazole resistance is present

The effects of fungicides used to control Rhynchosporium secalis in winter barley were investigated in five field trials in Co. Down, Northern Ireland. Benzimidazole resistance was present in the R. secalis population of each site. Fungicide performance was evaluated in terms of disease control, maintenance of green leaf area, grain yield and grain quality. Samples of R. secalis isolates taken before and after treatment in each field were assessed for sensitivity to carbendazim in the laboratory. Carbendazim contributed little to disease control, the maintenance of green leaf area, grain yield or grain quality. Carbendazim was associated with a significant increase in disease severity in some plots. Treatments containing carbendazim, either alone or in mixture, caused an increase in the proportion of carbendazim-resistant isolates within R. secalis populations. Propiconazole-containing treatments performed well in terms of disease control, maintenance of green leaf area and grain quality. They caused no change in carbendazim resistance frequency.     

Soft rot and blackleg reactions in diploid potato hybrids inoculated withErwinia spp.

From 1993 to 1996 three groups of potato genotypes were evaluated for resistance toErwinia spp.: (1) 31 interspecific diploid hybrids (28 resistant and three susceptible), (2) five hexaploid or pentaploid somatic hybrids ofSolanum tuberosum (tbr) ×S. brevidens (brd), and (3) eight cultivars. Two evaluation methods were applied: tuber point inoculation with eitherErwinia carotovora ssp.atroseptica (Eca) orE. chrysanthemi (Ech) to test tuber soft rot resistance and stem base inoculation with Eca to test blackleg resistance. Some resistant diploid hybrids and somatic hybrids oftbr × brd were superior to cultivars for both tuber and stem resistance. Tuber and stem resistance toErwinia spp. in the most resistant diploid hybrids were equal to the highly resistant somatic hybrids oftbr xbrd. Tuber resistance to Eca was highly correlated to tuber resistance to Ech (r=0.815***). In two years of evaluation for stem resistance, three diploid hybrids and a derivative of one of the somatic hybrids (USA M 264) failed to develop symptoms of blackleg following inoculation with Eca, Analyses of variance for tuber and stem resistance indicated significant effects of genotype, year and genotype × year interaction. A positive relationship between tuber and stem resistance toErwinia spp. has been found, however the genetic control of resistance in tuber and stem is partially independent. In the case of Eca the correlation coefficient was r = 0.725***. Therefore it should be possible to obtain resistant genotypes to both blackleg and tuber soft rot. Several resistant diploid hybrids were selected from among those tested, which also have several other characters desirable for potato breeders.     

PCR-RFLP detection of the E198A mutation conferring resistance to benzimidazoles in field isolates of Monilinia laxa from Greece

Sensitivity to benzimidazoles in isolates of the brown rot pathogen (Monilinia laxa) collected from stone fruit in central and northern Greece was evaluated and the molecular basis for resistance was investigated. M. laxa isolates were classified as benzimidazole – sensitive (S) or highly resistant (HR) based on their sensitivity profiles to carbendazim. Thirty seven percent of the isolates belonged to the HR phenotype, carried no apparent fitness penalties and exhibited resistance factor values (based on EC₅₀ values) greater than 500. Highly resistant isolates were also less sensitive to the benzimidazoles benomyl and thiophanate-methyl but more sensitive to the N-phenylcarbamate diethofencarb and the benzamide zoxamide compared to isolates belonging to the S phenotype. Fungitoxicity tests with fungicides belonging to other chemical classes revealed no cross resistance relationships between benzimidazoles and the dicarboximide iprodione, the phenylpyrrole fludioxonil, the hydroxyanilide fenhexamid, the carboxamide boscalid, the triazole tebuconazole and the strobilurin-type fungicide pyraclostrobin, indicating that a target site modification is probably responsible for the resistant phenotypes observed. Comparison of the β-tubulin gene DNA sequences between resistant and sensitive isolates revealed a point mutation resulting from the E198A substitution of the corresponding protein in all HR isolates tested. An Eco31I restriction site in the β-tubulin gene, which was destroyed in HR M. laxa isolates, allowed the development of a PCR-RFLP diagnostic for the detection of the E198A resistance mutation.     

Progress report: Resistance to the Colorado potato beetle and potato leafhopper inSolanum tuberosum subsp.andigena

An extensive collection of 1527 clones ofS. tuberosum subsp.andigena was screened in 1973 in unreplicated plots to test for resistance to the Colorado potato beetle and potato leafhopper. All clones were susceptible to the beetle. Of 17 clones selected in the 1973 tests for resistance to the potato leafhopper, 7 selections were significantly more resistant than the control (Katahdin) in a replicated experiment in 1974.     

Resistance of Botrytis cinerea Pers. to dicarboximide fungicides — a literature review

The dicarboximides (iprodione, procymidone, vinclozolin) are a relatively new group of protectant fungicides which are used mainly for the control of Botrytis cinerea Pers. The literature documenting the emergence of dicarboximide-resistant Botrytis strains and the current situation, is reviewed. Although dicarboximide-resistant Botrytis can be produced readily in the laboratory, extensive monitoring did not reveal the emergence of such strains in the field until after 2–3 years of intensive use of dicarboximides. Today, resistant strains are foubd in many areas where the dicarboximides have been in constant use. Nevertheless, loss of control by these fungicides has not, so far, become a major field problem, possibly because of a loss of vigour and pathogenicity that is often linked to dicarboximide resistance in Botrytis strains. Because resistant strains also produce varying numbers of sensitive conidia, such strains are rapidly resensitized in the absence of dicarboximide fungicides. These findings suggest that the nature of dicarboximide resistance is quite different to that of carbendazim resistance, in that carbendazim-resistant Botrytis strains show no loss of vigour and are, moreover, very stable.