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ABSTRACT Diaporthe phaseolorum and Phomopsis longicolla isolates from soybean were examined using traditional mycological characteristics and molecular methods. Cultural characteristics including types of fruiting bodies and conidia were assessed for isolates collected from soybean stems and seeds. Cultures were identified as P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, or D. phaseolorum var. sojae. Molecular markers for these groups were developed and analyzed using polymerase chain reaction restriction fragment length polymorphisms (PCR-RFLP) and DNA sequencing in the internal transcribed spacer (ITS) and the 5.8S ribosomal DNA. The ITS(4) and ITS(5) primers amplified PCR products for all isolates studied. Gel electrophoresis of undigested PCR products and DNA sequencing produced various fragment lengths including 604 bp for P. longicolla, 602 and 603 bp for D. phaseolorum var. caulivora, 603 bp for D. phaseolorum var. meridionalis, and from 597 to 609 bp for D. phaseolorum var. sojae. Digestion of these PCR products with enzymes AluI, HhaI, MseI, RsaI, and ScrFI resulted in distinct bands for identification of P. longicolla and the varieties of D. phaseolorum I. All P. longicolla, D. phaseolorum var. caulivora, and D. phaseolorum var. meridionalis isolates were distinguished using AluI and HhaI with RsaI or ScrFI. The banding patterns of D. phaseolorum var. sojae isolates were complex and were separated into 11 subgroups after digestion with AluI, HhaI, MseI, RsaI, and ScrFI. Phylogenetic analysis of 20 isolates of D. phaseolorum and P. longicolla based on the DNA sequence of the ITS region resolved six clades termed A, B, C, D, E, and F. Clade A included all sequenced D. phaseolorum var. caulivora isolates, two from Italy and one from the United States. Isolates in clade B were exclusively associated with D. phaseolorum var. meridionalis. Clades A and B formed a well-supported monophyletic group. Isolates in clades C, D, E, and F were morphologically defined as isolates of P. longicolla, D. phaseolorum var. sojae, and Diaporthe spp. The ITS sequences similarity of seven geographically diverse P. longi-colla isolates illustrated that P. longicolla isolates have a similar genetic background, with some affiliations to some D. phaseolorum var. sojae isolates. Morphological characteristics of the isolates along with the terminal clades of the ITS phylogeny suggest that P. longicolla is an individual species, D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis are varieties of D. phaseolorum, and D. phaseolorum var. sojae is either several varieties of D. phaseolorum or possibly several distinct species. 相似文献
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ABSTRACT Species-specific detection of Diaporthe phaseolorum and Phomopsis longicolla from soybean seeds was accomplished using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and TaqMan chemistry. To use these detection systems, fungal DNA was released from soybean seed coats using an ultrasonic processor to break the cells. DNA fragment lengths ranged from 200 to 1,200 base pairs (bp), with the majority of fragments <500 bp. Based on DNA sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA, three TaqMan primer/probe sets were designed. Primer/probe set PL-5 amplified a 96-bp fragment within the ITS1 region of P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, and D. phaseolorum var. sojae. Set PL-3 amplified a 86-bp DNA fragment within the ITS2 region of P. longicolla. Set DPC-3 amplified a 151-bp DNA fragment within the ITS2 region of D. phaseolorum var. caulivora. TaqMan primer/probe sets were able to detect as little as 0.15 fg (four copies) of plasmid DNA. When using PCR-RFLP for Diaporthe and Phomopsis detection, the sensitivity was as low as 100 pg of pure DNA. Among 13 soybean seed lots from Italy and the United States, the total Diaporthe and Phomopsis detected using a traditional seed-plating technique ranged from 0 to 32%. P. longicolla was most prevalent, followed by D. phaseolorum var. sojae. D. phaseolorum var. caulivora, which only occurred in 0.5% of the Italian seed lots, was not detected in the U.S. seed lots. D. phaseolorum var. meridionalis was not detected in either the U.S. or Italian seed lots. Using TaqMan primer/probe set PL-3, the frequency of P. longicolla was 18% in seed lot I3, similar to the frequency obtained from PCR-RFLP and potato dextrose agar plating detection. The frequencies of D. phaseolorum and P. longicolla in each seed lot obtained by the different detection methods were comparable with respect to total infection and individual species detection. However, TaqMan detection provided the fastest results of all the methods tested. 相似文献
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基于MAXENT的大豆南北方茎溃疡病菌在中国适生区的预测 总被引：4，自引：0，他引：4
利用MAXENT生态位模型和GIS系统,对大豆南北方茎溃疡病菌在我国的适生区进行预测.模型分析结果表明,这两种病菌在我国的潜在适生区域广泛.大豆北方茎溃疡病菌除了我国的宁夏、海南省外,其它省市均有该菌的适生分布区,其中适生等级高的地区主要在江苏、安徽、浙江、上海、江西和湖北6省.大豆南方茎溃疡病菌除了宁夏、青海、西藏外,我国的其它地区均为该菌的潜在适生区域,其中适生等级高的地区主要集中在江西、上海、江苏、安徽、浙江、河南以及陕西南部、四川东部和重庆的部分地区. 相似文献
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CLIMEX-GIS预测大豆北方茎溃疡病菌在中国的潜在分布 总被引：1，自引：0，他引：1
大豆北方茎溃疡病菌是大豆的重要病原菌,广泛分布于世界主要大豆产区,造成严重的产量和品质损失。本文应用生物模型CLIMEX结合GIS软件预测大豆北方茎溃疡病菌在中国的适生区,并根据EI值划分相应的适生等级。结果表明,大豆北方茎溃疡病菌在我国绝大部分地区适合生长,其中东北地区、华北地区和云贵高原地区处于中适生区或高适生区。该菌在我国还未报道,通过分析其在我国潜在分布区对于防止病菌的传入、传播和蔓延有重要的检疫意义。 相似文献
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The Rdm4 gene from soybean cv. Hutcheson has been extensively used to incorporate resistance to soybean stem canker (SSC), caused by Diaporthe phaseolorum var . meridionalis (Dpm), into soybean commercial cultivars. The objective of this work was to characterize the inheritance of the Rdm4 locus in different populations derived from the cross: J77-339 ( rdm / rdm , susceptible) × Hutcheson ( Rdm4 / Rdm4 , resistant) in independent interactions with two local isolates of Dpm. Four F2 populations were obtained and two were advanced to the F3 generation as separate F2:3 families to perform progeny tests. Each population was inoculated with the CE109 and/or CE112 isolates of Dpm. Within each plant–pathogen interaction, the resistance gene segregated as completely dominant. However, cross resistance, or opposite disease reactions, to CE109 and CE112 isolates of Dpm were observed in four F2:3 families, indicating an intergenic recombination event between two nonallelic genes interacting specifically with each isolate of Dpm. The distance between them, estimated as the recombination fraction, was 29%, suggesting that both genes were not tightly linked, but close enough to segregate together in most crosses. Results indicated the existence of a genomic region in cv. Hutcheson composed of race-specific resistance loci with at least two Rdm genes: the previously recognized Rdm4 and a novel gene, tentatively named Rdm5 , conferring specific resistance to Dpm isolates CE109 and CE112. 相似文献
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Inspection of 16 017 samples of soybean seeds imported into India from 1978 to 2004 resulted in the detection of 21 pathogens, including Peronospora manshurica which is not present in India. Seed-borne fungi of high economic significance included: Ascochyta sojicola , Botryotinia fuckeliana , Cercospora kikuchii , Colletotrichum dematium , Corynespora cassicola , Diaporthe phaseolorum var . sojae , Fusarium oxysporum, Glomerella cingulata , Glomerella glycines , Macrophomina phaseolina , Nectria haematococca , Passalora sojina , Thanatephorus cucumeris as well as other fungal pathogens for which soybean is not a host such as Alternaria padwickii , Cochliobolus sativus , Fusarium culmorum , Fusarium poae , Glomerella graminicola , Setosphaeria rostrata, Verticillium albo-atrum , etc. Some of the fungi detected have very wide host range. Details are presented on the fungi detected, the countries from which the imported consignments originated, and phytosanitary significance. 相似文献
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噻唑膦在不同介质不同pH条件下热贮稳定性 总被引：1，自引：0，他引：1
噻唑膦加工后的稳定性是剂型选择的关键,为了提高其稳定性,延长其持效期,本研究通过噻唑膦在几种介质中的热贮稳定性试验,研究了酸碱度及介质对噻唑膦化学稳定性及稳定剂环氧大豆油对噻唑膦水乳剂热贮稳定性的影响,从而探索出噻唑膦在不同介质中稳定的最佳pH范围。研究结果发现,噻唑膦在同一pH下不同介质中的稳定性表现为硅藻土膨润土水乳剂,当pH为4.5时噻唑膦的稳定性最佳,而且加入0.2%的环氧大豆油做稳定剂可使噻唑膦在水乳剂中的分解率控制在10%以下。总之噻唑膦在酸性介质中较稳定,在硅藻土和膨润土中的稳定性要高于水乳剂中的稳定性。 相似文献
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Says-Lesage V Roeckel-Drevet P Viguié A Tourvieille J Nicolas P de Labrouhe DT 《Phytopathology》2002,92(3):308-313
ABSTRACT Diaporthe/Phomopsis helianthi causes brown stem canker of sunflower (Helianthus annuus) and is responsible for considerable yield loss. This species shows considerable variation for morphological characters, growth, and pathogenicity. Molecular variability of two sample groups was assessed with amplified fragment length polymorphism (AFLP) markers. Isolates of the first sample were collected from infected sunflower tissues from the main regions in France where the crop is grown, whereas isolates from the second sample came from stems within a single field of sunflower. A soybean strain was taken as an outgroup for AFLP analyses. Within sample one, the greatest genetic distance among isolates was 0.97, whereas it was 0.44 within sample two isolates. For the whole of France, the average genetic distance was 0.68, whereas in the one field it was 0.12. Nei's genetic diversity indices were 0.20 and 0.06 for France and for one field, respectively. The greatest genetic distance was found between isolates from the most northern crops. The greatest genetic distance between D. helianthi isolates and the strain isolated from soybean was similar to that observed for D. helianthi isolates from different geographical areas. The problems in defining the genus Phomopsis are discussed. It is shown that internal transcribed spacer sequencing could be a useful criteria for Diaporthe/Phomopsis species determination. The considerable genetic variability of the pathogen could lead to the occurrence of new strains that could be more aggressive or more resistant to chemical control. 相似文献
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Gordon SG Kowitwanich K Pipatpongpinyo W St Martin SK Dorrance AE 《Phytopathology》2007,97(1):113-118
ABSTRACT Molecular analysis of sources of resistance to plant pathogens should expedite and confirm novel gene discovery and consequently the development of disease resistant cultivars. Recently, soybean plant introductions (PIs) were identified that contain putative novel Rps genes for resistance to Phytophthora sojae. The number of resistance genes that confer resistance to P. sojae isolates OH17 (1b,1d,2,3a,3b,3c,4,5,6,7) and OH25 (1a,1b,1c,1k,7) was then determined in several of the PIs. The objective of this study was to determine if the Rps genes present in these PIs were associated with eight described Rps loci that have been mapped on soybean molecular linkage groups F, G, J, and N. Nine F(2:3) soybean populations were genotyped with simple sequence repeat (SSR) markers linked to previously mapped Rps loci. The nine PI populations all had SSR markers associated (P < 0.01) with resistance to P. sojae isolate OH17 in the Rps1 region. Rps1c is a likely candidate in eight PIs but novel genes may also be possible, while novel genes may confer resistance in one PI to P. sojae isolate OHI7. Two or more Rps genes, including some that are potentially novel, confer resistance to P. sojae isolate OH25 in eight of the populations. However, based on the response to these two isolates, virulence already exists for at least some of the novel genes identified in this study. 相似文献
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María A. Chiesa Rosanna N. Pioli Mariana V. Cambursano Eligio N. Morandi 《European journal of plant pathology / European Foundation for Plant Pathology》2013,135(2):351-362
Soybean Stem Canker (SSC), caused by Diaporthe phaseolorum var. meridionalis (Dpm), is an important disease of soybean in Argentina. There are five known dominant genes that confer resistance to SSC, Rdm1 to Rdm5. Particularly, Rdm2 was identified in cv. Tracy-M and then it was stabilized in the breeding line T2. The Rdm4 gene was first identified in cv. Hutcheson. More recently it was found that this gene was linked to the Rdm5 gene, defining the Rdm4-5 resistance region in Hutcheson. The objective of this work was to analyze the behaviour of the dominant Rdm2, Rdm4 and Rdm5 genes interacting with the CE109 and CE112 local physiological races of Dpm, in different susceptible backgrounds (genotypes RA702 and J77-339). Rdm4 and Rdm5 segregated phenotypically as completely dominant genes in the specific interactions with the CE109 and CE112 isolates, respectively, in both susceptible backgrounds. Similarly, Rdm2 segregated as expected for a complete dominant gene in the specific interaction with the CE109 isolate, in both susceptible backgrounds. However, when interacting with the CE112 isolate, the Rdm2 gene did not segregate as expected for a completely dominant gene, neither in RA702 nor in J77-339 susceptible background. The distorted segregation of the Rdm2 gene was due to incomplete penetrance. To the best of our knowledge this is the first report documenting changes in the degree of penetrance of a soybean resistance gene (Rdm2) depending upon the physiological race of Dpm which interacts with and the genetic background in which the Rdm gene is being expressed. 相似文献
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The effect of two isolates of Chaetomium globosum (CgA-1, a producer and CgNA, a non-producer of antibiotic in vitro ) on the sporulation of pycnidia and survival of Diaporthe phaseolorum f. sp. meridionalis ( Dpm ) in soybean stems was determined under conditions of intermittent wetting and drying. Dpm -colonized stems placed on the surface of field soil or of quartz gravel were exposed to a 12- or 24-h wet period followed by a 48-h drying period until a cumulative wetness duration of 120, 240 or 480 h was attained. Both isolates reduced the pycnidia sporulation index to zero within 120 h of cumulative wetness duration if the wet period was 24 h. With a 12-h wet period the sporulation index decreased gradually, to between 4 and 8% after 480 h of cumulative wetness. In general, on the soil surface and with a 12-h wet period, CgA-1 reduced sporulation more efficiently than CgNA whereas with a 24-h wet period there was no difference between the isolates. Recovery of Dpm from C. globosum -treated stems decreased gradually with increase in wet period and cumulative wetness duration. The recovery rate from stems treated with isolate CgA-1 was always significantly less than that from stems treated with CgNA. In glasshouse experiments, disease incidence and the proportion of the stem length colonized by Dpm at the V6 growth stage was reduced significantly when soybean seeds treated with ascospores of C. globosum were sown in Dpm -infested soil. Incorporation of bran inoculum of either of the isolates in the upper soil layer dramatically reduced disease incidence. Isolate CgA-1 reduced the disease more efficiently than CgNA. 相似文献
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Bruna B. Brumer Valéria S. Lopes-Caitar Aline S. Chicowski Jessica D. Beloti Fernanda M. Castanho Danielle C. Gregório da Silva Sandremir de Carvalho Ivani O. N. Lopes Rafael M. Soares Claudine D. S. Seixas Ricardo V. Abdelnoor Francismar C. Marcelino-Guimarães 《European journal of plant pathology / European Foundation for Plant Pathology》2018,151(4):1009-1025
The complex of Diaporthe (asexual morph) species occurring on soybean constitutes an important pathogenic group associated with diseases such as pod and stem blight, seed decay and stem canker. Stem canker, caused by Diaporthe aspalathi, has been reported as the most aggressive form of canker and its occurrence has limited soybean crop productivity in the southern United States. The main form of pathogen control is the use of stem canker resistant soybean varieties. In this study, strains of Diaporthe and Phomopsis were isolated from stem and seeds of soybean in different locations in South America during the years 1989–2014. Genomic DNA from 26 isolates were analyzed by PCR-restriction fragment length polymorphism (RFLP) and Amplified Fragment Length Polymorphism (AFLP) techniques, and sequencing of internal transcribed spacer (ITS) regions of ribosomal DNA. The molecular analysis of ITS sequences by alignment with those of ex-type strains deposited in GenBank and morphological characteristics allowed the identification of Phomopsis longicolla, D. phaseolorum var. sojae, D. caulivora and D. aspalathi. An analysis of the pathogenicity of 13 isolates of D. aspalathi inoculated in soybean genotypes carrying different resistance genes to stem canker (Rdm1, Rdm2, Rdm3, Rdm4, Rdm5 and Rdm?) enabled us to identify the occurrence of at least three races of D. aspalathi occurring in Brazil. Among the isolates identified as D. aspalathi, both molecular and phenotypic analyses showed clustering depending on the date of collection and pathogenicity, which revealed the existence of variability of the pathogen. 相似文献
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大豆疫霉根腐病菌单游动孢子的毒性遗传与变异 总被引：7，自引：1，他引：6
采用离体叶柄伤口接种法测定大豆疫霉根腐病菌44号生理小种单游动孢子连续3代或4代分离后代的毒性,结果表明:从S1中选择与亲本相比毒性不发生变异的1号单游动孢子菌株(44号生理小种)和变异最大的30号单游动孢子菌株(1号生理小种)继续分离2代或3代,单游动孢子毒性变异趋势主要是从44号生理小种变异为3号生理小种,也有变异成毒性公式为7或1a,7的单游动孢子。大豆疫霉根腐病菌无性世代毒性变异几率很高,多数单游动孢子毒性在分离后代中都发生变异,产生不同的小种或毒性公式,并且毒性变异基本不能稳定遗传。 相似文献
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ABSTRACT Phytophthora sojae, which causes Phytophthora root and stem rot of soybean, is a serious disease worldwide and is managed primarily by deploying cultivars with resistance. Thirty-two soybean plant introductions (PIs), all but three of which were from South Korea, were proposed as new sources of single-gene resistance to P. sojae. The objective of this study was to characterize the inheritance of resistance to P. sojae in these PIs. Twenty-two soybean populations from crosses of these PIs and the susceptible cv. Williams were inoculated with P. sojae OH17 (vir 1b, 1d, 2, 3a, 3b, 3c, 4, 5, 6, 7), and OH25 (vir 1a, 1b, 1c, 1k, 7). These isolates were selected because they are virulent on soybeans with all known Rps genes and many Rps gene combinations. Thirteen of the twenty-two populations had consistent segregation responses following inoculations between the two generations. In two PIs, resistance was conferred by two genes to OH17 and three genes to OH25. Resistance to both isolates was conferred by a single gene in PI 398440 although the individual families were not resistant to the same isolates. The data suggest that six of the populations have three-Rps gene combinations as previously proposed, while another four may have either a novel Rps gene or a four-Rps gene combination. Based on this phenotypic analysis, novel and uncharacterized Rps genes may be present in this material. More importantly, these PIs may serve as sources of novel Rps genes that can be used to more effectively manage Phytophthora root and stem rot. 相似文献