Field evaluation of mungbeans for resistance to Cercospora leaf spot and powdery mildew

Abstract

Cercospora leaf spot, caused by Cercospora canescens and powdery mildew, caused by Erysiphe polygoni are two of the most important fungal diseases of mungbeans. Nearly 4000 accessions of the global mungbean collection at the Asian Vegetable Research and Development Center were screened for resistance to these two pathogens. Less than 4% and 12% of the accessions showed resistance to Cercospora leaf spot and powdery mildew, respectively. The level of resistance to Cercospora leaf spot varied and highly resistant lines were not found. A few lines were rated highly resistant to powdery mildew over several years but others were rated moderately resistant or susceptible in other years.     

Cryptic diversity,multilocus phylogeny,and pathogenicity of cercosporoid fungi associated with common bean and cowpea

In recent years, common bean (Phaseolus vulgaris) and cowpea (Vigna unguiculata) plants in the north of Iran have exhibited symptoms resembling Cercospora leaf spot (CLS) disease. This study was initiated to elucidate the taxonomy and pathogenicity of cercosporoid taxa associated with leaf spot diseases of these two legume crops in Iran. A total of 138 samples with CLS symptoms were collected from cultivated common bean and cowpea species in northern Iran and subjected to microscopic examination, resulting in identification of 98 Cercospora and 59 Pseudocercospora samples. A six-locus phylogenetic analysis (ITS, actA, tef1, gapdh, his3, and cmdA) coupled with examination of the morphology of 42 representative isolates from these samples confirmed that several cercosporoid fungi occur on common bean and cowpea in Iran. Five Cercospora species (C. iranica, C. cf. flagellaris, Cercospora sp. G, Cercospora sp. T, and C. vignigena) and two Pseudocercospora species (P. griseola f. griseola and P. cf. cruenta) were found; of these, C. cf. flagellaris was the dominant species, occurring on both common bean and cowpea. Pathogenicity tests confirmed that all seven species could infect leaves of common bean and/or cowpea. This is the first report of C. iranica, Cercospora sp. G, and Cercospora sp. T associated with common bean and/or cowpea in the world. In addition, C. vignigena was recorded for the first time in Iran. Results achieved in this study will assist strategies for the management of CLS disease of common bean and cowpea.     

Spatial pattern of Cercospora leaf spot of sugar beet in fields in long- and recently-established areas

Spatial disease pattern of Cercospora beticola was characterised during natural epidemics of Cercospora leaf spot (CLS) in sugar beet. We applied linear regression and geostatistical analyses to characterise CLS spatial patterns in three field trials, in long-established and recently-established CLS-areas, during two consecutive years. Linear regression showed a positive influence of average disease severity of within-row neighbouring plants (0.38 < < 0.88). Semi-variograms modelled the spatial dependence of disease severity for two directions per week in both years. Disease severity displayed strong spatial dependence over time. The within-row spatial dependence was the largest, but across-row dependence was irregular and weaker. Both long- and recently established areas showed strong spatial dependence of disease severity within row, decrease in variability between years and within the second trial year and a relation between and the relative nugget. Observed differences were more field than area specific. These spatial and temporal analyses indicated that disease severities of adjacent plants were dependent; hence, we concluded that C. beticola is dispersed mainly over short distances from plant to plant.     

Root infection of sugar beet by <Emphasis Type="Italic">Cercospora beticola</Emphasis> in a climate chamber and in the field

Sugar beet root infection by Cercospora beticola, the causal agent of Cercospora leaf spot (CLS), was studied in a climate chamber and in the field. In the climate chamber, root incubation of susceptible seedlings with a conidial suspension resulted in disease incidences that were significantly different for two sugar beet cultivars (Auris: 0.8 ± 0.14 and A00170: 0.5 ± 0.18; P < 0.05) with regard to the control treatment 35 days after root incubation in a standard potting soil-fine river sand mixture. In a field trial with susceptible cv. Savannah with soil-incorporated CLS-infested leaf material, disease developed four weeks earlier in the infested plots than in the control plots. The probability that disease develops in the field was significantly higher for the infested than for the control plots (P < 0.05). Symptomless plants from infested field plots transferred to the glasshouse to induce leaf spot symptoms showed a significantly higher probability to induce symptom development (0.4 ± 0.08), than plants from control plots (0.02 ± 0.02) (P <0.05) 14 days after transfer. This probability was significantly higher than for plants that remained in three of the infested field plots (0.2 ± 0.04; 0.2 ± 0.05 and 0.2 ± 0.04 respectively), except for one infested field plot (0.4 ± 0.05) on July 5. We conclude that C. beticola is able to infect sugar beet seedlings through their roots and that latent CLS infections in sugar beet lead to symptom development at high temperatures (> 20 °C) and high relative humidity (> 95) in our climate chamber or after canopy closure in the field. Quantification of root infection and long term survival in soil is necessary to assess its contribution to the epidemiology and life cycle of Cercospora beticola. Cultural methods such as a wider crop rotation, management of crop debris and ploughing systems may provide control strategies alternative to or reducing fungicide input.     

Identification of the G143A mutation associated with QoI resistance in Cercospora beticola field isolates from Michigan,United States

BACKGROUND: Cercospora leaf spot (CLS), caused by the fungus Cercospora beticola, is the most serious foliar disease of sugar beet (Beta vulgaris L.) worldwide. Disease control is mainly achieved by timely fungicide applications. In 2011, CLS control failures were reported in spite of application of quinone outside inhibitor (QoI) fungicide in several counties in Michigan, United States. The purpose of this study was to confirm the resistant phenotype and identify the molecular basis for QoI resistance of Michigan C. beticola isolates. RESULTS: Isolates collected in Michigan in 1998 and 1999 that had no previous exposure to the QoI fungicides trifloxystrobin or pyraclostrobin exhibited QoI EC₅₀ values of ?0.006 µg mL^?1. In contrast, all isolates obtained in 2011 exhibited EC₅₀ values of > 0.92 µg mL^?1 to both fungicides and harbored a mutation in cytochrome b (cytb) that led to an amino acid exchange from glycine to alanine at position 143 (G143A) compared with baseline QoI‐sensitive isolates. Microsatellite analysis of the isolates suggested that QoI resistance emerged independently in multiple genotypic backgrounds at multiple locations. A real‐time PCR assay utilizing dual‐labeled fluorogenic probes was developed to detect and differentiate QoI‐resistant isolates harboring the G143A mutation from sensitive isolates. CONCLUSION: The G143A mutation in cytb is associated with QoI resistance in C. beticola. Accurate monitoring of this mutation will be essential for fungicide resistance management in this pathosystem. Copyright © 2012 Society of Chemical Industry     

Lack of influence of host plant disease resistance on the evolution of resistance to sterol demethylation-inhibiting (DMI) fungicides in<Emphasis Type="Italic">Cercospora beticola</Emphasis>

Field experiments were conducted during 1997 and 1998 to determine the effects of sugar beet cultivar susceptibility to Cercospora leaf-spot on the sensitivity ofCercospora beticola isolates to the triazole fungicide flutriafol. Four cultivars with different levels of disease resistance were treated in experimental plots with six spray applications of flutriafol. Disease assessments were carried out at 15-day intervals. Sensitivity to flutriafol was measured on isolates collected from the plots ∼15 days after the last flutriafol application. Measurements of disease severity and calculations of AUDPC (area under disease progress curve) values showed a distinct differentiation among cultivars, reflecting their level of disease resistance. Disease severity was significantly lower in cvs. ‘Bianca’ and ‘Areth’ than in ‘Univers’ and ‘Rizor’ both in the untreated and in the flutriafol-treated plots. Fungal isolates from flutriafol-treated plots were less sensitive to the fungicide than were isolates from untreated plots. However, no differences in isolate sensitivity were observed among the cultivars, as regards their level of disease resistance. Despite the fact that the use of resistant cultivars cannot eliminate selectively the resistant strains, it can eliminate both resistant and sensitive isolates. Reducing the number of treatments with DMIs, by applying them only when environmental conditions are favorable for disease development, is a prerequisite for successful resistance management; therefore, the use of disease-resistant varieties could aid toward management of DMIs resistance inC. beticola. http://www.phytoparasitica.org posting May 6, 2003.     

Interactive Effects of Host, Pathogen and Mineral Nutrition on Grey Leaf Spot Epidemics in Uganda

Grey leaf spot incited by Cercospora zeae-maydis is a new devastating foliar disease of maize in East Africa. For effective control, elucidation of the most critical elements of the grey leaf spot disease pyramid is important. This study investigated the role of mineral nutrition, pathogen variability and host resistance in the epidemic. Trials were conducted under field and controlled environments. The 28 isolates used in the controlled environment varied significantly (P 0.05) in parasitic fitness measured indirectly as disease efficiency, but no infection pattern could be attributed to known C. zeae-maydis pathotypes. Data from field trials showed that host resistance and mineral nutrition significantly (P 0.05) affected disease efficiency, with highest disease development occurring in nitrogen-augmented plots. Exclusive phosphorus application had no clear effect on grey leaf spot epidemics but combined application with nitrogen significantly (P 0.05) reduced the predisposition effects of nitrogen to the disease. Overall, treated plots had less disease than unfertilised plots. Fertiliser application had no effect on sporulation capacity, while cultivars significantly affected it. Geographic differences in amount of disease were observed, suggesting environment influences on grey leaf spot incidence. The results suggest that the current grey leaf spot epidemics in East Africa are due to favourable cultivars, poor mineral nutrition and environmental interactions.     

Dynamics of cercospora leaf spot disease determined by aerial spore dispersal in artificially inoculated sugar beet fields

Cercospora beticola is one of the most important fungal pathogens of sugar beet, causing cercospora leaf spot (CLS) disease. Due to the decreasing efficacy of various fungicides caused by resistance traits, the development of a sustainable disease management strategy has become more important. Therefore, detailed knowledge about the epidemiology of the pathogen is crucial. Until now, little was known about the spatiotemporal dispersal of C. beticola spores from the primary inoculum source. Rapid detection of C. beticola spores could facilitate a more precise and targeted disease control. Therefore, a TaqMan real-time PCR assay for detection and quantification of C. beticola spores caught with Rotorod spore traps was established. In 2016 and 2017, field trials were conducted to monitor C. beticola aerial spore dispersal and disease development within an inoculated field and in the adjacent noninoculated area. With the established detection method, C. beticola spores were successfully quantified and used as a measure for aerial spore dispersal intensity. The analysis of the spatiotemporal spread of C. beticola spores revealed a delay and decrease of aerial spore dispersal with increasing distance from the inoculated area. Consequently, disease incidence and severity were reduced in a similar manner. These results imply that spore dispersal occurs mainly on a small scale within a field, although long distances can be overcome by C. beticola spores. Moreover, secondary aerial spore dispersal from sporulating leaf spots seems to be the main driver for CLS disease development. These results provide an important basis for further improvement of CLS control strategies.     

玉米抵御玉蜀黍尾孢菌侵入的生理机制

为深入探讨玉米抗灰斑病的机制,以抗/感玉米灰斑病自交系78599-1、OH43Ht N和掖478、K12为材料,采用比色皿法和RT-PCR法相结合研究了接种玉蜀黍尾孢菌毒素后玉米叶片中防御酶活性、丙二醛(MDA)含量及防御酶合成相关基因SOD、CAT、APX和GR的表达量。结果显示,接种该病菌毒素后,抗、感材料叶片中防御酶活性均升高,多数在接种7 d时达到峰值,且抗病材料防御酶活性峰值高于感病材料;供试材料叶片中MDA含量均降低,且抗病材料低于感病材料;供试材料叶片中SOD、CAT和GR的表达量均上升,其中CAT的表达量在78599-1、掖478和K12接种5d时达到峰值,灰度值分别为228.67、161.33和178.00,与其酶活性变化趋势一致;SOD和GR的变化规律与其酶活性变化不一致;APX的表达量仅在OH43Ht N中上升,接种7 d后达到峰值。表明抗病材料调控防御酶活性的能力强,防御酶基因的表达与其酶活性变化存在关联性。     

The effect of timing of fungicide applications on control of frogeye leaf spot and grain yield of soybeans

Soybean cultivar Samsoy 1, and the breeding lines TGx 849-313D and TGx 996-26E, grown in a field with a heavy epidemic of frogeye leaf spot caused byCercospora sojina, were treated with double foliar applications of the fungicide benomyl. The treatments were made using four application schedules at six different growth stages, starting from V₃ (fully developed leaves, beginning with trifoliate nodes) to R₅ (beginning seed_, to determine the effect of the fungucide timing on frogeye leaf spot severity, soybean grain yield and grain quality. Generally, applications at R₁ (beginning bloom) and R₃ (beginning pod) significantly (P<-0.05) reduced disease severity in the 2 susceptible genotypes, Samsoy 1 and TGx 849-313D. Plot yields of these genotypes were also significantly greater than the untreated controls when the fungicide applications were made at R₁ and R₃. There was no significant difference in diseave severity or grain yield, between the untreated control and the different times of application, on the resistant genotype TGx 996-26E. Improved seed germination and lower levels of seed infection byC. sojina occurred for all fungicide timings in the susceptible genotypes. The results suggest that fungicide spraying initiated at R₁ and followed up at R₃ is most effective in frogeye leaf spot control and can also result in higher grain yields, than applications made earlier or later in the season. Control of frogeye leaf spot, however, is best achieved by growing resistant cultivars.     

Resistance in leaf blades assessed by counting conidia correlates with whole-plant-specific resistance in leaf sheaths in a compatible rice–Magnaporthe oryzae interaction

Resistance in the leaf blades of rice plants against a virulent race of the rice blast fungus Magnaporthe oryzae was quantitatively examined using a modified spot inoculation method. Numbers of conidia produced in the lesions were affected by plant age and paralleled the frequency of resistance infection types, which is indicative of whole-plant-specific resistance (WPSR), in the inoculated leaf sheaths of the corresponding plants. Exogenous abscisic acid treatment, which suppresses WPSR, also increased the susceptibility of the leaf blades. These results indicate a correlation between the resistance of the leaf blades and the WPSR in the leaf sheaths.     

<Emphasis Type="Italic">Cercospora zeina</Emphasis> is the causal agent of grey leaf spot disease of maize in southern Africa

The aim of our study was to identify the causal agent of grey leaf spot disease of maize in southern Africa. Single-conidial cultures were recovered from maize leaves with typical disease symptoms sampled from several fields in South Africa, Zambia and Zimbabwe. Morphology, cultural characteristics, and a PCR-based test using Cercospora zeae-maydis and C. zeina-specific primer sets identified all single-conidial cultures as C. zeina. In addition, sequence alignment of DNA fragments of the internal transcribed spacer region (ITS1, ITS2, and the 5.8S gene) and elongation factor 1-α grouped all cultures in the same clade as the C. zeina ex-type culture CBS 118820. To by-pass cultivation of the slow-growing fungus, a rapid method to isolate DNA directly from lesions was successfully applied for PCR identification of C. zeina with species-specific ITS and histone primers. Koch’s postulates were fulfilled for C. zeina by artificially inoculating maize plants in a greenhouse, re-isolating conidia emerging from lesions and verifying pathogen identity with molecular techniques. These results provide evidence that confirms the presence of C. zeina and absence of C. zeae-maydis in commercial maize plantations in southern Africa.     

棉花对硃砂叶螨抗性的鉴定和机制研究

1988-1991年进行的棉花抗硃砂叶螨试验表明,海岛棉和二倍体栽培棉(中棉和草棉)对硃砂叶螨具有较高水平的抗性,它们的抗螨性已通过远缘杂交技术转移到了感螨的陆地棉栽培品种中。来自四川省和河南省的一些品种以及来自美国的PD种质系对硃砂叶螨的抗性比湖北省当地品种要差。叶片有毛,无腺体和高含淀粉的棉花,硃砂叶螨为害较重;而叶片具有致密茸毛、棉酚、可溶性糖、叶绿素和类黄酮含量高以及叶片鲜重高的品种抗螨性较强。     

Identification of a new pathotype of Puccinia hordei with virulence for the resistance gene Rph7

Barley leaf rust resistance gene Rph7, derived from barley accession Cebada Capa, is the most effective R-gene for resistance to Puccinia hordei. Virulence for this gene was known in the USA, Israel and Morocco but not yet in Europe. We found an unexpected leaf rust infection in the field at Córdoba, Spain in 2004 on Rph7 carrying lines. This virulence for Rph7 was confirmed in growth chamber experiments, being the first report of Rph7 virulence in European populations of P. hordei. A collection of 680 barley accessions was screened for resistance against this new isolate. Twelve accessions showed segregation with individual plants showing resistance based on hypersensitivity (low infection type). These individual resistant plants were selected and grown in the greenhouse to obtain seeds.     

烟粉虱传播的番茄褪绿病毒和番茄黄化曲叶病毒对不同番茄品种的复合侵染

为明确烟粉虱传播的番茄褪绿病毒(Tomato chlorosis virus,ToCV)与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)对不同番茄品种的复合侵染情况,于2015年11月在山东省寿光市温室内采集13个番茄品种共390份疑似发病植株叶片,对不同番茄品种的TYLCV抗性和2种病毒的复合侵染以及温室内发病番茄植株上烟粉虱成虫的带毒率进行检测。结果表明,采集的13个番茄品种经分子标记检测鉴定均为TYLCV杂合抗性;不同番茄品种ToCV与TYLCV的复合侵染率存在明显差异,大果番茄粉宴和贝瑞上复合侵染率最高可达73.3%,而樱桃番茄八喜上未检测到这2种病毒的复合侵染。此外,在发病番茄植株上采集的烟粉虱成虫体内可检测到2种病毒,其中烟粉虱ToCV带毒率为90.7%,TYLCV带毒率为80.0%,同时检测到ToCV与TYLCV的概率为71.3%。表明ToCV和TYLCV的复合侵染在山东省番茄生产中普遍发生,烟粉虱可同时携带这2种病毒并广泛传播。     

The rapid and accurate determination of germ tube emergence site by Blumeria graminis conidia

The development of appressoria by germinating Blumeria graminis conidia depends on its germ tubes making contact with the host surface. Low angle, low temperature scanning electron microscopy showed that 80% or more of first-formed germ tubes of f. spp. hordei, tritici and avenae conidia emerged from close to the host leaf surface, and so made contact with it allowing them to become functional primary germ tubes. Light microscopy of f. sp.hordei confirmed this result and, in addition, showed that germ tubes frequently emerged close to, and contacted, various hydrophobic and hydrophilic artificial substrata. Geometric models of conidium-substratum interfaces were developed and a “surface point method” was derived to predict the frequency with which contact would result if germ tube emergence was a random phenomenon. However, observed contact frequencies were far higher (c. three to eight times) than predicted. Thus, the germ tube emergence site was determined as a response to substratum contact. In part, this appeared to be a non-specific response. Nevertheless, germ tube contact frequencies were greater on the curved surface of leaf epidermal cells than on planar surface, suggesting that specific recognition of leaf surface characteristics may promote directional emergence. The area of contact required to stimulate directional germ tube emergence was very small: contact with a microneedle tip or with a spiders’ suspension thread was sufficient for many conidia. Similarly, on leaves, the only contact is between the tips of a limited number of conidial wall projections and the edges of epicuticular leaf wax plates. Micromanipulation to roll conidia so that their original site of contact with a leaf was rotated away from it, led to the majority of first-formed germ tubes growing away from the leaf, i.e. emerging close to the site of original contact. The experiments indicated that the site of germ tube emergence is determined within 1 min of deposition. This implicated the release of conidial extracellular materials in recognition of the conidium-leaf surface contact site.     

基于高通量测序检测甜菜生尾孢中真菌病毒多样性

为明确甜菜生尾孢Cercospora beticola中真菌病毒的多样性,2020年自新疆维吾尔族自治区、内蒙古自治区、黑龙江省和北京市采集感染褐斑病甜菜病叶,鉴定获得78株甜菜生尾孢菌株,运用高通量的宏转录组测序技术检测甜菜生尾孢所携带的真菌病毒种类;将分离到的甜菜生尾孢按照菌落生长表型分为4组,进行宏转录组测序,将得到的序列经过拼接、聚类和基因功能注释,筛选出真菌病毒相关同源序列。序列分析结果显示,4组样品共比对到6个真菌病毒科,包括双分体病毒科Partitiviridae、双核糖核酸病毒科Birnaviridae、低毒病毒科Hypoviridae、裸露RNA病毒科Narnaviridae、泛欧尔密病毒科Botourmiaviridae和转座病毒科Metaviridae,以及未分科的负义单链RNA病毒。研究结果丰富了甜菜生尾孢真菌病毒资源库。     

Development and Evaluation of an in vitro Detached Leaf Assay forc Pre-Screening Resistance to Fusarium Head Blight in Wheat

An in vitro detached leaf assay, involving the inoculation of detached leaves with Microdochium nivale, was further developed and used to compare with whole plant resistance ratings to Fusarium head blight (FHB) of 22 commercial cultivars and published information on 21 wheat genotypes, identified as potential sources for FHB resistance. An incubation temperature of 10 °C and isolates of M. nivale var. majus of intermediate pathogenicity were found to be the most suitable for the differential expression of several components of partial disease resistance (PDR), namely incubation period, latent period and lesion length, in wheat genotypes used in the detached leaf assay. There were highly significant differences (P < 0.001) for each component of PDR within commercial cultivars and CIMMYT genotypes. Positive correlations were found between incubation period and latent period (r = 0.606; P < 0.001 and r = 0.498; P < 0.001, respectively, for commercial cultivars and CIMMYT genotypes), inverse correlations between incubation period and lesion length (r = -0.466; P < 0.01 and r = –0.685; P < 0.001, respectively) and latent period and lesion length (r = –0.825; P < 0.001 and r = –0.848; P < 0.001, respectively). Spearman rank correlations between individual PDR components and UK 2003 recommended list ratings were significant for incubation period (r_s = 0.53; P < 0.05) and latent period (r_s = 0.70; P < 0.01) but not for lesion length (r _s = –0.26). Commercial cultivars identified with high resistances across all three PDR components in the detached leaf assay also had high whole plant FHB resistance ratings, with the exception of cv. Tanker which is more susceptible than the results of the detached leaf assay suggested, indicating an additional susceptibility factor could be present. Agreement between resistances found in the detached leaf assay and resistance to FHB suggests resistances detected in detached leaves are under the same genetic control as much of the resistances expressed in the wheat head of the commercial cultivars evaluated. In contrast, high resistances in each of the PDR components were associated with higher susceptibility across 19 CIMMYT genotypes previously evaluated as potential breeding sources of FHB resistance (incubation period: r = 0.52; P < 0.01, latent period: r = 0.53; P < 0.01, lesion length: r = –0.49; P < 0.01). In particular, the CIMMYT genotypes E2 and E12 together with Summai #3, known to have high levels of whole plant FHB resistance, showed low levels of resistance in each PDR component in the detached leaf assay. Such whole plant resistances, which are highly effective and not detected by the detached leaf assay, do not appear to be present in Irish and UK commercial cultivars. The most resistant Irish and UK commercial cultivars were comparable to the genotype Frontana and the most resistant CIMMYT germplasm evaluated in the leaf assay.     

Relating plant and pathogen development to optimise fungicide control of phoma stem canker (<Emphasis Type="Italic">Leptosphaeria maculans</Emphasis>) on winter oilseed rape (<Emphasis Type="Italic">Brassica</Emphasis><Emphasis Type="Italic">napus</Emphasis>)

In winter oilseed rape experiments at Rothamsted in 2000/01 to 2002/03 growing seasons, the severity of phoma stem canker epidemics in summer depended on the timing of phoma leaf spot epidemics in the previous autumn, and hence on the timing of Leptosphaeria maculans ascospore release. The first major release of L. maculans ascospores was earlier in 2000 (26 September) and 2001 (18 September) than in 2002 (21 October). Consequently, the autumn phoma leaf spot epidemic was also earlier in 2000 and 2001 than in 2002. The resulting stem canker epidemics were severe by harvest (July) in 2001 and 2002 but not in 2003. No correlation was found between the severity or duration of phoma leaf spotting (lesion days or lesion °C-days) and the subsequent severity of phoma stem canker epidemics. Rates of leaf production and loss were similar in the three growing seasons. Out of ca. 25 leaves produced on plants during each season, leaf numbers 10–14 generally remained on plants for the longest. Treatment with flusilazole + carbendazim in autumn decreased the severity of phoma leaf spotting for several weeks after treatment, decreased the severity of stem canker the following summer and increased yield significantly in 2001 and 2002 but not in 2003. The most effective timings for flusilazole + carbendazim application were when leaves 7–11 were present on most plants and at least 10% of plants were affected by phoma leaf spot. Two half-dose applications of fungicide reduced phoma stem canker and increased yield more than a single full dose application when phoma leaf spot epidemics were early (<800 °C-days after sowing).     

Effects of thiophanate-methyl and azoxystrobin on the composition of Cercospora kikuchii populations with thiophanate-methyl-resistant strains

Azoxystrobin was recently registered in Japan for the control of purple seed stain of soybean caused by Cercospora kikuchii, because the pathogen has developed resistance to thiophanate-methyl. To investigate the effects of these fungicides on the frequency of C. kikuchii strains resistant to thiophanate-methyl and on the genotype structure of the population, we sowed purple-stained seeds, approximately 40% of which were infected with resistant strains, as inocula with asymptomatic seeds and applied thiophanate-methyl and azoxystrobin during the reproductive growth of soybeans. The isolation frequency of resistant strains increased more than 99% by thiophanate-methyl but was not significantly increased by azoxystrobin. In amplified fragment length polymorphism (AFLP) DNA fingerprinting, genotypic diversity was significantly decreased by thiophanate-methyl but was not affected by azoxystrobin. In addition, the similarity of the AFLP genotype structure was increased by thiophanate-methyl but not by azoxystrobin. These results suggest that thiophanate-methyl selectively inhibited the proliferation of sensitive strains, which resulted in a small number of genotypes, most of which were resistant strains. Azoxystrobin was found to nonselectively inhibit proliferation of the pathogen, which retained a large number of genotypes including thiophanate-methyl-sensitive or thiophanate-methyl-resistant strains or both. The nucleotide sequence data for the cytochrome b gene are available in the DDBJ/EMBL/GenBank databases under accession number AB231863.