Metalaxyl sensitivity of isolates of Phytophthora erythroseptica in Prince Edward Island

Isolates of Phytophthora erythroseptica , causal agent of pink rot of potato, were obtained from diseased tubers collected in Prince Edward Island (PEI), Canada. In an in vitro assay of field and single-zoospore isolates, all isolates of the pathogen from PEI were sensitive to metalaxyl (EC₅₀ < 0·5  µ g mL⁻¹), unlike moderately or highly resistant reference isolates obtained from Maine, USA. Allozyme-banding patterns at the glucose-6-phosphate isomerase ( Gpi ) locus were identical (91/91) for all isolates of P. erythroseptica examined from PEI and Maine, but could be used to distinguish isolates of P. erythroseptica from local isolates of Phytophthora infestans . Inoculation of potato tubers from plants treated with foliar applications of metalaxyl in the field indicated that compounds inhibitory to metalaxyl-sensitive isolates of P. erythroseptica were present in the periderm, even after 4 months' storage. By contrast, moderately or highly resistant isolates of the pathogen caused significantly ( P  ≤ 0·05) more disease than sensitive isolates in tubers from plants treated with metalaxyl. The effectiveness of metalaxyl in controlling pink rot in a particular region will depend on resistance levels of local populations of P. erythroseptica . Preventing the development and spread of resistant strains of P. erythroseptica will be critical in maintaining metalaxyl as an effective chemical for control of this pathogen.     

贵州省烟草黑胫病菌对甲霜灵的抗药性

为了明确贵州地区烟草黑胫病菌对甲霜灵的抗药性水平, 从贵州省12个市、县采集黑胫病病株, 经分离纯化鉴定, 获得98株黑胫病菌株, 采用生长速率法测定其对甲霜灵的敏感性。结果表明, 采自不同地区的菌株对甲霜灵的敏感性差异较大, 各供试菌株的EC50 值分布范围为0.419 2～20.486 1 μg/mL; 各地区平均EC50 在0.551 9～10.986 0 μg/mL之间; 最低抗药性水平为1.00, 最高抗药性水平为48.87, 相差48.87倍, 各地区平均抗药性水平在1.32～26.21之间。遵义、兴仁、贵定、赫章和兴义有抗甲霜灵菌株出现, 除遵义部分菌株的抗性水平表现为高抗外, 其余菌株对甲霜灵产生的抗性均处于中抗水平。     

六省市致病疫霉交配型及其对几种杀菌剂的敏感性

测定了2000-2002年采自四川、重庆、吉林、黑龙江、福建、北京六省(市)致病疫霉菌株的交配型及其对几种杀菌剂的敏感性。结果显示,来自六省市的74个菌株中73株为典型的A1交配型,1株采自四川的菌株为可自育的A1交配型,未发现A2交配型。测定的37株代表菌中,对甲霜灵表现敏感、中抗、高抗的分别占27.0％、27.0％和46.0％。吉林、福建、黑龙江及重庆都存在高抗菌株。研究中还发现甲霜灵可促进部分抗性菌株的菌丝生长。随机抽取15个对甲霜灵表现敏感、中抗和高抗的菌株,测定其对几种杀菌剂的敏感性,结果全部对腈嘧菌酯和烯酰吗啉表现敏感;其中2株对丙酰胺表现敏感,11株表现中抗,2株表现高抗。结果也显示:烯酰吗啉和腈嘧菌酯与甲霜灵之间不存在交互抗性,具有应用潜力。     

Selection for decreased sensitivity to phosphite in Phytophthora cinnamomi with prolonged use of fungicide

To test the hypothesis that resistance in Phytophthora cinnamomi to control by the fungicide phosphite (phosphonate) would arise in sites with prolonged use of phosphite, 30 P. cinnamomi isolates were collected from a range of sites with different phosphite‐use histories, including phosphite‐treated and untreated avocado orchards, and phosphite‐treated and untreated native vegetation sites. The colonizing ability of these isolates was tested by different inoculation methods against a range of host tissues, treated and untreated with phosphite, including mycelial stem inoculation on clonally propagated Leucadendron sp., mycelial root inoculation of lupin seedlings and zoospore inoculation of Eucalyptus sieberi cotyledons. Isolates from avocado orchards with a long history of phosphite use were, on average, more extensive colonizers of the phosphite‐treated Leucadendron sp., lupin seedling roots and Eucalyptus sieberi cotyledons. These isolates did not colonize untreated plant tissue (Leucadendron sp.) more extensively than isolates from sites with no history of phosphite use and no isolates were resistant to control by phosphite. Analysis of all isolates with microsatellite markers revealed the majority were from a single clonal lineage. Selection for decreased sensitivity to phosphite in planta has taken place within asexual clonal lineages of P. cinnamomi in sites with prolonged use of phosphite.     

In vitro somatic growth and reproduction of phenylamide-resistant and -sensitive isolates of Phytophthora erythroseptica from infected potato tubers in Idaho

Pink rot of potato, most commonly caused by Phytophthora erythroseptica , is a major field and post-harvest problem in southern Idaho, USA, particularly since 1998 when isolates resistant to the phenylamide fungicide metalaxyl-M (mefenoxam) were detected. Isolates of P. erythroseptica were collected from infected tubers in 2001 and 2002 from six Idaho counties and tested for resistance to metalaxyl-M on amended agar. Metalaxyl-M resistant (MR) and metalaxyl-M-sensitive (MS) isolates were identified in six counties; 160 isolates were highly resistant, seven moderately resistant and 57 sensitive to metalaxyl-M with mean EC₅₀ values of 182, 23 and 0·5 mg L⁻¹ ai metalaxyl-M, respectively. Mycelial growth rates and oospore production in agar were assessed for 20 MS and 20 MR isolates at 10, 15, 20, 25 and 30°C. Growth rates of MR isolates were between 2·5 and 3·1 times greater ( P  < 0·05) than those of MS isolates at 10, 15, 20 and 25°C, and oospore production was between 6·8 and 20·5 times greater ( P  < 0·0001) for MR than for MS isolates at the same temperatures. Colony growth in V8 broth at 18°C was greater for MR than MS isolates ( P  < 0·0032). However, zoospore production at 18°C was greater for MS than for MR isolates ( P  < 0·0109), and zoospore production m m ⁻¹ of colony circumference was also greater for MS than for MR isolates, 14 191 and 9959, respectively ( P  = 0·0109). Sexual reproduction of MR isolates in nature may be greater than MS isolates, but MS isolates may be more asexually fit based on the fitness parameters studied.     

北方五省(区)马铃薯晚疫病菌对甲霜灵和精甲霜灵的敏感性检测

为了明确我国不同地区马铃薯晚疫病菌对甲霜灵和精甲霜灵的抗性状况,2007-2009年从河北、黑龙江、吉林、辽宁和内蒙古马铃薯主产区采集马铃薯晚疫病菌,采用菌丝生长速率法和叶盘漂浮法测定其对甲霜灵的敏感性,采用叶盘漂浮法测定其对精甲霜灵的敏感性.结果表明:以菌丝生长速率法检测的380株晚疫病菌中和以叶盘漂浮法检测的222株晚疫病菌中对甲霜灵的抗性菌株分别占80％和73.5％;菌丝生长速率法检测结果显示,河北省对甲霜灵的抗性菌株所占频率从2007年的100％降为2008年的66.4％,2009年又回升至74.2％,而吉林和黑龙江两省对甲霜灵的抗性菌株分布频率呈上升趋势.叶盘漂浮法检测的95个菌株中对精甲霜灵的抗性菌株占54.5％,其中河北省40个菌株对精甲霜灵的中间型菌株占优势(62.5％),抗性菌株仅占34.9％,而其他4省55个菌株对精甲霜灵的抗性菌株占优势(69.1％).受检测的北方五省(区)马铃薯晚疫病菌群体中对甲霜灵和精甲霜灵的抗性菌株已占优势,马铃薯晚疫病菌已普遍对甲霜灵及精甲霜灵产生抗性.在对甲霜灵和精甲霜灵普遍产生抗性的地区,应优先选用与甲霜灵、精甲霜灵作用机理不同的药剂防治马铃薯晚疫病.     

Genetic diversity,sensitivity to phenylamide fungicides and aggressiveness of Phytophthora ramorum on Camellia,Rhododendron and Viburnum plants in Spain

Phytophthora ramorum has been detected in official plant health surveys on Rhododendron, Viburnum and Camellia in ornamental nurseries in northern Spain since 2003. A collection of 94 isolates of P. ramorum was obtained from 2003 to 2008 from plants with symptoms at different geographical locations. Isolates were identified based on morphology and sequence of the rDNA ITS region. Mating type, genetic variation, sensitivity to phenylamide fungicides and aggressiveness of these isolates were determined. All isolates belonged to the A1 mating type, ruling out the possibility of genetic recombination. Seven microsatellite markers were used to study genetic diversity; three out of the seven microsatellite markers were polymorphic within the Spanish population of P. ramorum. This study confirms that all Spanish isolates of P. ramorum belonged to the EU1 lineage. Twelve intralineage genotypes were detected, five that are unique to Spain (EU1MG38, EU1MG41, EU1MG37, EU1MG39 and EU1MG40) and seven that are also present in at least one other European country (EU1MG1, EU1MG29, EU1MG22, EU1MG13, EU1MG2, EU1MG18 and EU1MG26). Genotypes EU1MG37, EU1MG39 and EU1MG40 were isolated from Rhododendron from one region; EU1MG38 and EU1MG41 were isolated from Camellia from two different regions. Isolates of genotype EU1MG38 were resistant to metalaxyl and mefenoxam. The level of genetic diversity within the Spanish population of P. ramorum is limited and indicates a relatively recent clonal expansion.     

泸州烟草黑胫病病原生理小种组成及对甲霜灵和烯酰吗啉的敏感性

烟草黑胫病(Phytophthora nicotianae)是四川省泸州烟区最为严重的病害之一,为明确泸州烟草黑胫病生理小种组成及其对甲霜灵和烯酰吗啉的敏感性,从泸州主产烟区采集样品,分离得到24株烟草黑胫病菌株;通过鉴别寄主法结合TTZ生化反应,发现24株菌株中,0号生理小种15株,占62.50%,1号生理小种9株,占37.50%。对24株菌株进行甲霜灵和烯酰吗啉的药剂敏感性检测,发现甲霜灵对24株菌株的EC50范围在0.052~0.513μg/mL之间,敏感菌株和中抗菌株分别占41.67%和58.33%,未发现高抗菌株;烯酰吗啉对24株菌株的EC50范围在0.110~1.615μg/mL之间,敏感菌株和中抗菌株分别占37.50%和62.50%,未发现高抗菌株。上述结果表明,泸州烟区烟草黑胫病菌同时存在0号生理小种和1号小种,而且该烟区的黑胫病菌已对甲霜灵和烯酰吗啉产生不同程度的抗性。     

Evaluating diverse systems of tobacco genetic resistance to Phytophthora nicotianae in Yunnan,China

Black shank, caused by the soilborne pathogen Phytophthora nicotianae, is one of the most devastating diseases affecting tobacco production in China. The most effective strategy for reducing economic loss from this pathogen is development and use of resistant tobacco varieties. Multiple sources and systems of resistance have been developed in the Western Hemisphere; however, populations of P. nicotianae are variable around the world, including the predominance of different races. Different P. nicotianae isolates may react differently on tobacco plants with different systems of resistance, a possibility that could complicate the breeding of cultivars with resistance that is effective in different tobacco production regions worldwide. The objective of this research was to evaluate an array of tobacco germplasm possessing different systems of genetic resistance to black shank disease in tobacco-growing regions of Yunnan, China. Resistance types included simply inherited resistance mechanisms introgressed from wild Nicotiana relatives and polygenic partial resistance systems of N. tabacum origin. The loci of Wz exhibited high level resistance to black shank in the five diverse disease environments in Yunnan, China. K326 Php/−Wz/− genotype and Beinhart 1000 exhibited the greatest levels of resistance in both 2015 and 2016. Field observed results for 13 tobacco genotypes were highly correlated with those tested in growth chamber evaluation. These findings suggest that both Wz− and Beinhart 1000-mediated resistance have important commercial value in flue-cured tobacco breeding programmes in China. Cultivars developed for black shank resistance in China may also have utility in other tobacco-growing areas.     

Sensitivity of Phytophthora infestans to flumorph: in vitro determination of baseline sensitivity and the risk of resistance

The sensitivity of 127 Phytophthora infestans isolates to flumorph was determined in 2003 and 2004. The isolates originated from two geographical regions and showed similar levels of sensitivity in both years. Baseline sensitivities were distributed as a unimodal curve with EC₅₀ values for growth of mycelia ranging from 0·1016 to 0·3228  µ g mL⁻¹, with a mean of 0·1813 (± 0·0405) µ g mL⁻¹. There was no cross-resistance between flumorph and metalaxyl. Laboratory studies were conducted to evaluate the risk of P. infestans developing resistance to flumorph. Mutants resistant to metalaxyl or flumorph were obtained by treating mycelium of wild-type isolates with ultraviolet radiation. Metalaxyl-resistant mutants were obtained with a high frequency and exhibited resistance factor values (EC₅₀ resistant/EC₅₀ sensitive phenotypes) of more than 100, while flumorph-resistant mutants were obtained at much lower frequencies and had very small resistance factors (1·5–3·2). There was cross-resistance between flumorph and dimethomorph, but not with azoxystrobin or cymoxanil. Most flumorph-resistant mutants showed decreases in hyphal growth in vitro and in sporulation both in vitro and on detached leaf tissues. These studies suggested that the risk of resistance developing was much lower for flumorph than metalaxyl. However, as P. infestans is a high-risk pathogen, appropriate precautions against resistance development should be taken.     

Monitoring and characterization of Magnaporthe grisea isolates with decreased sensitivity to scytalone dehydratase inhibitors

Rice blast fungus isolates were collected in Kyushu to investigate resistance to scytalone dehydratase inhibitors of melanin biosynthesis (MBI-D). In 2001, failure of control of rice blast was reported in the Saga prefecture, where MBI-Ds have been used since 1998. At that time, the distribution of resistant isolates was mainly limited to that area. However, in 2002, resistant isolates were detected in all prefectures of Kyushu. DNA fingerprinting analysis showed that the mutation causing resistance to MBI-Ds had arisen independently in each area. These data suggest that resistant isolates may occur in any area and become dominant under continuous selection pressure for MBI-Ds. Nevertheless, resistant strains can be controlled by reductase inhibitors of melanin biosynthesis (MBI-R) or commercial rice seed disinfectants.     

Analysis of Colletotrichum musae populations from Brazil reveals the presence of isolates with high competitive ability and reduced sensitivity to postharvest fungicides

In this study, the sensitivity of 218 isolates of Colletotrichum musae to imazalil and thiabendazole was evaluated, as well the fitness and competitive ability of less sensitive isolates. There was a positive correlation between the sensitivity to the two fungicides, but the isolates were more sensitive to imazalil. The estimated effective concentration of the fungicide able to inhibit mycelial growth by 50% (EC₅₀) was used to select four isolates with the lowest and the highest values for both fungicides, which were considered as sensitive (S) and less sensitive (LS), respectively. The level of sensitivity was maintained after 10 successive transfers on fungicide-free medium. Both fungicides were effective in controlling the disease caused by S isolates of C. musae in detached banana fruit when recommended doses were used. However, only imazalil was able to control the disease caused by LS isolates. For both fungicides, analysis of fitness-related variables (mycelial growth, sporulation, germination, and virulence) showed no difference between the groups of S and LS isolates, but a large variation was observed within the group. The LS isolates to thiabendazole that showed a mutation (F200Y) in the β-tubulin gene did not have fitness penalties. Our results allow a better understanding of the sensitivity and fitness of isolates of C. musae from Brazil, and demonstrate the importance of periodic monitoring to determine the frequency of LS isolates in populations, aiming at more effective management of anthracnose in banana orchards in Brazil.     

中国4省份葡萄霜霉病菌群体对烯酰吗啉的抗性及适合度

为明确我国山西、山东、河北和云南省葡萄霜霉病菌对烯酰吗啉的抗性以及田间病菌群体的适合度,本研究采用Taqman-MGB技术检测了2020年-2021年采自上述4省份292株葡萄霜霉病菌对烯酰吗啉的抗性频率;采用叶盘法测定了田间抗性菌株与敏感菌株的适合度。结果表明,上述4个地区的平均烯酰吗啉抗性频率及抗性等位基因频率分别为64.0%和74.7%,其中山东省蓬莱区最高(91.2%和96.3%),河北省廊坊市(73.8%和85.0%)和云南省宾川县(75.0%和82.6%)次之,山西省清徐县(16.7%和34.7%)最低。适合度分析发现,田间抗性菌株的产孢能力明显高于敏感菌株,但二者的复合适合度指数无显著差异。上述结果表明,4省份霜霉病菌群体均对烯酰吗啉产生了不同程度的抗性,田间抗性菌株与敏感菌株相比无明显适合度变化。     

Phenotypic and genotypic diversity of Phytophthora infestans populations in Scotland (1995–97)

In a survey of Scottish potato late blight ( Phytophthora infestans ) populations from 1995 to 1997, nearly 500 isolates were collected from over 80 disease outbreaks in commercial potato crops and gardens/allotments. The isolates were characterized by mating type, resistance to the fungicide metalaxyl and almost 300 were examined by DNA-based AFLP fingerprinting. These data were examined alongside cropping details to determine the population structure in the context of existing disease management strategies. A1 and A2 mating type isolates were present in both commercial potato crops and gardens or allotments although they coexisted more frequently in the latter sites. One-fifth of the isolates collected were of the A2 mating type and the frequency was similar over the 3 years and amongst sites. In 1995 the proportions of isolates that were sensitive and resistant to metalaxyl were equal (∼40%) but, over the following 2 years, the frequency of resistant isolates decreased and that of intermediate isolates increased. The mating type response to metalaxyl differed markedly, with 52% of A1 and only 5% of A2 isolates being resistant. Considerable molecular diversity was observed, with over half of the isolates having unique AFLP patterns. Analysis of the molecular and phenotypic data revealed a broad clustering of the population into three groups. Many factors point to an A2 population restricted by its sensitivity to phenylamides. The majority of the A2 isolates were found in a single AFLP group, but the presence of mixed mating type samples, an increasing frequency of isolates of intermediate metalaxyl resistance and the extent of the AFLP diversity suggest occasional sexual recombination, and thus gene flow, between groups.     

Occurrence of Corynespora cassiicola isolates resistant to boscalid on cucumber in Ibaraki Prefecture, Japan

A total of 651 isolates of cucumber corynespora leaf spot fungus ( Corynespora cassiicola ) collected from cucumber in Japan, either with (438 isolates) or without (213 isolates) a prior history of boscalid use, were tested for their sensitivity to boscalid by using a mycelial growth inhibition method on YBA agar medium. Additionally, seven isolates of C. cassiicola obtained from tomato, soybean, eggplant (aubergine) and cowpea in different locations in Japan were tested before boscalid registration. Minimum inhibitory concentration (MIC) and 50% effective concentration (EC₅₀) values for 220 isolates from crops without a prior history of boscalid use ranged from 0·5 to 7·5 μg mL⁻¹ and from 0·04 to 0·59 μg mL⁻¹, respectively. Two hundred and fourteen out of 438 isolates collected from ten cucumber greenhouses in Ibaraki Prefecture, Japan, which received boscalid spray applications showed boscalid resistance, with MIC values higher than 30 μg mL⁻¹. Moreover, resistant isolates were divided into two groups: a moderately resistant (MR) group consisting of 189 isolates with EC₅₀ values ranging from 1·1 to 6·3 μg mL⁻¹, and a very highly resistant (VHR) group consisting of 25 isolates with EC₅₀ values higher than 24·8 μg mL⁻¹. MR isolates were detected from all ten greenhouses, but VHR isolates were detected from only three. As a result of fungus inoculation tests which used potted cucumber plants, control failures of boscalid were observed against resistant isolates. Efficacy of boscalid was remarkably low against VHR isolates in particular. This is the first known report on boscalid resistance in Japan.     

北京地区草莓灰霉病菌对异菌脲的抗性及抗性分子机制

由灰葡萄孢(Botrytis cinerea Pers.)引起的草莓灰霉病是草莓上危害严重的病害之一。为了解北京地区草莓灰霉病菌对二甲酰亚胺类常用杀菌剂异菌脲的抗性,本研究采用最低抑制浓度法分别检测了2013年和2014年从北京地区15个草莓园采集的共计121株草莓灰霉病菌对异菌脲的抗性。结果表明,北京地区草莓灰霉病菌对异菌脲存在较高的抗性频率,2014年较2013年的抗性频率略有上升,由40.4%上升为45.3%。不同草莓园菌株的抗性频率差异很大,可能与用药水平有关。2014年的低抗、中抗和高抗菌株数分别占检测菌株总数的9.4%、28.1%和7.8%。利用PCR技术扩增编码组氨酸激酶基因BcOS1中与二甲酰亚胺抗性相关的区段,对抗性菌株的分子机制进行了初步研究,结果表明BcOS1基因第1 214位核苷酸发生了2类突变:以第Ⅰ类为主,菌株的抗性水平为中抗和高抗,由ATC突变为AGC,导致第365位氨基酸由异亮氨酸变为丝氨酸;第Ⅱ类菌株为低抗,由ATC突变AAC,导致第365位氨基酸由异亮氨酸变为天冬酰胺。     

Sterol composition of isolates of Erysiphe graminis f.sp. tritici differing in sensitivity to fenpropimorph

Isolates of Erysiphe graminis f.sp. tritici with wild-type or reduced sensitivity to fenpropimorph were similar in sterol composition, viz. ergosta-5,24(28)-dienol (±90%) and episterol (±10%). Following treatment with fenpropimorph, the relative content of episterol increased in conidia of all isolates tested, while that of ergosta-5,24(28)-dienol decreased. These results suggest that fenpropimorph, under the test conditions used, does not inhibit activity of sterol Δ¹⁴-reductase or Δ⁸→Δ⁷-isomerase but probably interferes with the final part of the demethyl sterol synthesis. However, modifications in this part of the pathway are probably not responsible for the decreased sensitivity of the pathogen to the fungicide. © 1998 SCI.     

Sensitivity to silthiofam, tebuconazole and difenoconazole of Gaeumannomyces graminis var. tritici isolates from China

BACKGROUND: Wheat take‐all caused by Gaeumannomyces graminis var. tritici (Ggt) has become an emerging threat to wheat production in the last few years. Silthiofam is very effective against Ggt, and recently it has been widely used for the control of take‐all in China. However, farmers have noted a decline in control efficacy with this compound in some wheat fields, suggesting that the pathogen may have developed resistance to silthiofam. RESULTS: Of the 66 Ggt isolates collected from different locations in China, 27 were resistant to silthiofam. There was no cross‐resistance between silthiofam and tecuconazole or difenoconazole. The effectiveness of silthiofam in controlling take‐all was compromised on wheat inoculated with silthiofam‐resistant isolates. Based on the DNA fingerprinting generated by microsatellite PCR, two predominant genetic clusters were found among these isolates and were clearly associated with the sensitivity to silthiofam. CONCLUSION: Silthiofam has a high risk in the development of resistance in Ggt. Tebuconazole and difenoconazole show great potential for control of take‐all on wheat. Results from this study provide useful information for take‐all control and the management of fungicide resistance. Copyright © 2012 Society of Chemical Industry     

Sensitivity of Podosphaera aphanis isolates to DMI fungicides: distribution and reduced cross‐sensitivity

BACKGROUND: Management of strawberry powdery mildew, Podopshaera aphanis (Wallr.), requires numerous fungicide treatments. Limiting epidemics is heavily dependent on sterol demethylation inhibitors (DMIs) such as myclobutanil or penconazole. Recently, a noticeable reduction in the efficacy of these triazole fungicides was reported by strawberry growers in France. The goal of this study was to investigate the state of DMI sensitivity of French P. aphanis and provide tools for improved pest management. RESULTS: Using leaf disc sporulation assays, sensitivity to myclobutanil and penconazole of 23 isolates of P. aphanis was monitored. Myclobutanil EC₅₀ ranged from less than 0.1 to 14.67 mg L^?1 and for penconazole from 0.04 to 4.2 mg L^?1. A cross‐analysis and a Venn diagram showed that there was reduced sensitivity and a positive correlation between the less sensitive myclobutanil and penconazole isolates; 73.9% of isolates were less sensitive to a DMI and 47.8% exhibited less sensitivity to both fungicides. CONCLUSION: The results show that sensitivity to myclobutanil and, to a lesser extent, penconazole has become less efficient in strawberry powdery mildew in France. Therefore, urgent action is required in order to document its appearance and optimise methods of control. Copyright © 2009 Society of Chemical Industry