Partial resistance to Wheat dwarf virus in winter wheat cultivars

Four Hungarian winter wheat cultivars were investigated for their susceptibility to the geminivirus Wheat dwarf virus (WDV). Previously, two cultivars (Mv Regiment and Mv Emese) were assessed by breeders to exhibit virus symptoms in the field, whereas Mv Dalma and Mv Vekni showed few symptoms. Two inoculation techniques for WDV, vector transmission with the leafhopper Psammotettix alienus and agroinoculation, were used. Leafhopper transmission was more efficient than agroinoculation. However, irrespective of the technique used, no Mv Dalma or Mv Vekni plants showed clear WDV symptoms. In contrast, 3/30 Mv Emese and 4/36 Mv Regiment plants showed dwarfing and chlorosis after agroinoculation and 13/17 and 14/15 plants, respectively, had clear WDV symptoms after vector transmission. WDV‐specific PCR showed that Mv Vekni and Mv Dalma plants could be infected, especially following vector transmission (approximately 50% infection), but at significantly lower frequency than Mv Emese or Mv Regiment plants (100% infection). Furthermore, real‐time PCR showed that WDV DNA accumulated to much lower levels in infected Mv Vekni and Mv Dalma plants than in infected Mv Regiment and Mv Emese plants. The data strongly suggest that Mv Vekni and Mv Dalma are partially resistant to WDV infection. As WDV resistance has not previously been identified in wheat, and because WDV can cause significant yield losses, the resistance of Mv Vekni or Mv Dalma will provide a valuable breeding resource.     

Studies on the host range of the barley strain of Wheat dwarf virus using an agroinfectious viral clone

Comparative analysis of the host ranges of the barley and wheat strains of Wheat dwarf virus (WDV; family Geminiviridae ; genus Mastrevirus ) in Europe has been severely hampered by the lack of an infectious clone of the barley strain. To remedy this situation an agroinfectious clone of a Hungarian isolate of the barley strain (WDV-Bar[HU]) was constructed and its virulence tested in barley ( Hordeum vulgare ), wheat ( Triticum aestivum ), rye ( Secale cereale ) and oat ( Avena sativa ) by agroinoculation. Although all four species could be systemically infected by the isolate, infections were asymptomatic in the rye and oat cultivars tested. WDV-Bar[HU] induced chlorosis and stunting symptoms typical of WDV in barley, while in wheat low infection rates but high mortality of infected seedlings were observed. In contrast, a much higher percentage of wheat plants agroinoculated with a wheat-strain isolate (WDV-[Enk1]) became systemically infected. WDV-[Enk1] in wheat caused symptoms similar to those caused by WDV-Bar[HU] in barley. WDV-Bar[HU] was leafhopper-transmissible to barley seedlings, in which it caused typical WDV symptoms; geminate virus particles were isolated from the infected leaves. Comparison of the genomic sequences of 11 barley strain isolates from Europe and Turkey revealed that whereas WDV-Bar[HU] represents a typical barley-strain isolate that is not detectably recombinant, the Turkish barley isolate (WDV-Bar[TR]) is probably a recombinant between a barley-strain isolate and an as-yet-undescribed WDV-like mastrevirus species.     

Phylogenetic and serological analyses reveal genetic diversity of Agrobacterium vitis strains in Japan

Twenty-eight strains of Agrobacterium vitis , including both tumorigenic and nonpathogenic phenotypes involving 26 isolated in Japan and strains NCPPB 3554^T and NCPPB 2562 isolated in Australia and Greece, respectively, were characterized by means of a slide agglutination test (SAT) using antisera prepared against somatic antigens. Phylogenetic analyses were also carried out, using the results of repetitive sequence-based polymerase chain reaction and the partial nucleotide sequences of pyrG , recA and rpoD . The A. vitis strains separated into four serogroups (A to D) based on the SAT reactivity. The phylogenetic analyses showed A. vitis strains separated into four tumorigenic groups (A to D) and one nonpathogenic group (E). Serogroups A to C corresponded exactly to genetic groups A to C, respectively, whereas serogroup D further divided into two distinct genetic groups, D and E. Genetic group E was isolated in Okayama Prefecture, Japan, and all strains of it have been found to coexist with tumorigenic strains belonging to the other groups within the same galled grapevine tissues. These results suggest that A. vitis strains are genetically heterogeneous and can be separated into several genetic groups. The differences between the nucleotide sequences of pyrG , recA and rpoD of the genetic groups will enable the development of a simple and convenient monitoring method, which will increase understanding of the dynamics of each genetic group of A. vitis in the natural environment.     

Geographic distribution and genetic diversity of Fusarium graminearum and F. asiaticum on wheat spikes throughout China

A large number of Fusarium graminearum and F. asiaticum isolates were collected from wheat spikes from all regions in China with a history of fusarium head blight (FHB) epidemics. Isolates were analysed to investigate their genetic diversity and geographic distribution. Sequence characterized amplified region (SCAR) analyses of 437 isolates resolved both species, with 21% being F. graminearum (SCAR type 1) and 79% being F. asiaticum (SCAR type 5). AFLP profiles clearly resolved two groups, A and B, that were completely congruent with both species. However, more diversity was detected by AFLP, revealing several subgroups within each group. In many cases, even for isolates from the same district, AFLP haplotypes differed markedly. Phylogenetic analyses of multilocus DNA sequence data indicated that all isolates of SCAR type 1, AFLP group A were F. graminearum , whilst isolates of SCAR type 5, AFLP group B were F. asiaticum , demonstrating that it is an efficient method for differentiating these two species. Both species seem to have different geographic distributions within China. Fusarium graminearum was mainly obtained from wheat growing in the cooler regions where the annual average temperature was 15°C or lower. In contrast, the vast majority of F. asiaticum isolates were collected from wheat growing in the warmer regions where the annual average temperature is above 15°C and where FHB epidemics occur most frequently. This is the first report of the distribution of, and genetic diversity within, F. graminearum and F. asiaticum on wheat spikes throughout China.     

Genetic diversity of the wheat yellow rust population in Pakistan and its relationship with host resistance

Yellow rust, caused by Puccinia striiformis f.sp. tritici (PST), is an important disease that threatens wheat production in Pakistan. This study was designed to explore the virulence and simple sequence repeat (SSR) diversity of the Pakistani PST population and the ongoing selective pressures of widely grown wheat cultivars. Analyses of 49 isolates sampled from the North‐West Frontier Province of Pakistan led to the identification of 12 distinct pathotypes. The virulence frequencies of v2 (virulent against Yr2), v6, v7, v9, vSU and v27 ranged from 63% to 100%. Virulences v3, v4, v17 and vSD were uncommon, whilst v5, v10, v15, v24, v32 and vSp were not detected. The pathotypes thus described were then classified into 27 distinct genotypes. Bayesian structure analysis clustered these genotypes into five groups (in addition to one hybrid isolate) which represent three distinct lineages of the SSR‐based phylogenetic tree. Of the studied isolates, 80%, represented by three predominant pathotypes (P1–P3), belonged to the same characteristic Pakistani lineage, whilst the other isolates were close to either a Mediterranean lineage or a Northern European lineage. Genetic recombination was detected within P2 isolates. Resistance genes postulated in 40 Pakistani wheat cultivars indicated the high frequency of Yr2, Yr6, Yr7, Yr9, Yr27 and YrSU. Only 11 cultivars were found to be resistant to P1–P3. Migration and varietal diversity factors might contribute to maintaining the currently high genetic diversity in Pakistani PST, and have serious regional implications for wheat improvement programmes.     

Effect of combining two genes for partial resistance to Barley yellow dwarf virus-PAV (BYDV-PAV) derived from Thinopyrum intermedium in wheat

In this study two sources of resistance to Barley yellow dwarf virus (BYDV), both originating from Thinopyrum intermedium , were combined in a single genotype, line ZT, using GISH (genomic in situ hybridization) and a new molecular marker of the partial resistance gene Bdv2 . Susceptible wheat cv. Sunstar, the translocation line TC14 carrying Bdv2 , the addition line ZH with the group-2 chromosome arm carrying resistance, and line ZT with both resistances were inoculated with five strains of Barley yellow dwarf virus -PAV. The tests confirmed the resistance of TC14 and ZH and revealed an additive effect of the two sources of resistance in ZT. The resistance of line ZT was characterized by a proportion of infected plants significantly lower than the parental lines TC14 and ZH (42% vs. a mean of 76% for the parents) and a very low virus titre (area under the virus concentration progress curve of 1·2 vs. a mean on 6·3 for the parents).     

山茶褪绿矮缩伴随病毒杭州分离物的序列和进化分析

Through sequencing and assembly of small RNAs, a Camellia chlorotic dwarf-associated virus (CaCDaV) was identified on a camellia plant showing chlorosis and distorting leaves from Hangzhou (CaCDaV-HZ1). The full genome of CaCDaV-HZ1 was determined and compared with that of a CaCDaV isolate from Chongqing (CaCDaV-CA1). Results showed that they shared 85.7% overall nucleotide sequence identity. Insert and delete mutations were mainly distributed in the non-coding region, whereas the coding regions mainly contained synonymous substitution. All genes except V3 have identical length among the two isolates. Additionally, the small RNA profile of CaCDaV-HZ1 was also analysed and results showed that 24 nt is the major class of viral-derived siRNA indicating that methylation is a major resistance mechanism of camellia plants against geminiviruses.     

山茶褪绿矮缩伴随病毒杭州分离物的序列和进化分析

<正>双生病毒(Geminivirus)是一类基因组为单链环状DNA的植物病毒,具有孪生的病毒粒子。国际病毒分类委员会(International Committee on Taxonomy of Viruses,ICTV)最新的分类报告将双生病毒分为9个属[1]。随着测序技术的发展,利用小RNA或转录组测序可鉴定植物中的已知和未知病毒,新的双生病毒也不断被发现和鉴定[2]。山茶褪绿矮缩伴随病毒(Camellia chlorotic dwar     

BYDV PREDICTOR: a simulation model to predict aphid arrival, epidemics of Barley yellow dwarf virus and yield losses in wheat crops in a Mediterranean-type environment

BYDV PREDICTOR, a simulation model, was developed to forecast aphid outbreaks and Barley yellow dwarf virus (BYDV) epidemics in wheat crops in the grainbelt region of southwest Australia, which has a Mediterranean-type climate. The model used daily rainfall and mean temperature to predict aphid ( Rhopalosiphum padi ) buildup in each locality before the commencement of the cereal-growing season in late autumn, and to forecast the timing of aphid immigration into crops. The introduction of BYDV by aphid immigrants, aphid buildup within the crop, spread of BYDV, and yield losses were predicted for different sowing dates. The model simulations were validated with 10 years' field data from five different sites in the grainbelt, representing a wide range of scenarios. When first aphid arrival dates ranging from 1 June to 2 September were compared with predictions, 65% of the variation between sites and years was explained. Progress curves for the predicted percentage of plants infected with the serotype BYDV-PAV closely resembled the starting point and shape of those recorded in 14 out of 18 scenarios. Sensitivity analysis confirmed that the combination of a high proportion of immigrants vectoring BYDV, early sowing of crops and early start to aphid arrival relative to sowing date led to the most BYDV spread and greatest yield loss. The model was incorporated into a decision support system used by farmers in targeting sprays against aphids to prevent virus spread in autumn and winter. BYDV PREDICTOR could serve as a template for modelling similar virus/aphid vector pathosystems in other regions of the world, especially those with Mediterranean-type climates.     

Genetic diversity of Melon necrotic spot virus and Olpidium isolates from different origins

The geographic incidence, genetic diversity and phylogenetic relationships of Melon necrotic spot virus (MNSV) and Olpidium isolates were studied in three cucurbit species from several Latin American and European countries on different collecting dates. Of the 112 cucurbit samples analysed, 69 from Guatemala, Honduras, Mexico, Panama and Spain were DAS‐ELISA‐positive for MNSV. Olpidium bornovanus and O. virulentus infections, and MNSV infections mixed with these Olpidium species, were observed for all these countries. Twenty‐nine MNSV isolates from all the origins where the virus was detected were selected and amplified by RT‐PCR. The resulting RT‐PCR of the p29, p89, p7A, p7B and p42 proteins was used to estimate the genetic diversity and the phylogenetic relationships of the MNSV population. The sequences obtained in this study were compared with the MNSV sequences of the NCBI database, and three groups were recovered by nucleotide composition according to geographical origins: the EU‐LA genotype group (with two subgroups: EU and LA, European and Latin American isolates, respectively), the JP melon genotype group (Japanese melon reference isolates) and the JP watermelon genotype group (Japanese watermelon reference isolates). The genetic diversity in the entire p7A and p7B proteins of MNSV suggests that these coding regions are under strong selective pressure. Additionally, the rDNA‐ITS region was analysed in 40 O. bornovanus and O. virulentus isolates associated with each geographical location and host examined. Phylogenetic analysis showed two groups for each Olpidium species, and these groupings were related to the host from which they were originally isolated.     

Effects on growth of wheat plants of isolates of Gaeumannomyces/Phialophora-complex fungi in different conditions of soil moisture, temperature, and photoperiod

The effects on spring wheat (Triticum aestivum L., cv. Mario) of nine isolates of the Gaeumannomyces/Phialophora complex, ranging from non-pathogenic to pathogenic, were studied under different conditions of soil moisture, soil temperature, and photoperiod in growth chambers which simulated different autumn weather conditions. The experimental conditions were based on data (e.g. temperature) from representative sites (loamy sand, Muencheberg, Northeast Germany) collected in the last three decades. The results of seedling inoculation tests for four non-pathogenic isolates were partly in agreement with results from field trials done over 4 years. One non-pathogenic G. graminis var. tritici isolate (G 33) increased consistently dry weight of shoots in the simulation, and grain yield in field experiments. For non-pathogenic isolates, warm temperatures with moderate soil moisture most often stimulated plant growth, with less effect in cold dry soil conditions. The decrease in seedling growth caused by pathogenic isolates was influenced only slightly by temperature changes, but was often enhanced by increased soil moisture.     

Relative importance of different types of inoculum to the establishment of Mycosphaerella graminicola in wheat crops in north‐west Europe

The contribution of wheat debris to the early stages of septoria leaf blotch epidemics was assessed in a 3‐year field experiment. First lesions were detected very early (December) in the case of an early sowing (mid‐October), showing that the first contamination could occur as soon as the seedlings emerge. The tested debris management options (chopped debris, removal of debris followed by tillage, or tillage in absence of debris) had a strong effect, although transient, on the epidemic dynamic: the more debris present on the soil surface, the more severe initial disease was. The magnitude of differences between treatments differed substantially between years. The relative production of pycnidiospores and ascospores was measured on the chopped debris. Peaks in pycnidiospore and ascospore production coincided in October–November. Both types of spores can be involved as primary inoculum in north‐west European conditions. The local amount of pycnidiospores available on debris in the field, estimated per square metre, was 1000‐fold the local ascospore production. Moreover, inoculum production was quantified on debris exposed to different environmental conditions. Autumnal conditions, characterized by moderate temperature with alternating wet and dry periods, were favourable for the production of both pycnidiospores and ascospores, as shown by the high inoculum production on debris exposed to field or outdoor conditions. By late autumn, the canopy became the most important source of pycnidiospores, and this period, characterized by the decreasing role of debris as a local source of inoculum compared to distant potential sources, can be considered as the end of the early epidemic stages.     

陕西韩城严重发生的小麦矮缩病病原鉴定与原因分析

2007年在陕西韩城发现一种新的小麦病毒病害,症状表现为严重矮缩、黄化、条斑和分蘖增多等,发病面积约0.07万hm²,病田减产达50%,严重地块甚至绝收。本研究通过对采集自我国陕西韩城的7个样品进行PCR鉴定、全基因组序列测定及比较,证实陕西韩城样品确是小麦矮缩病毒(WDV)侵染所致,并对发病原因及其流行趋势进行了分析。这是小麦矮缩病在我国麦田大发生的首次报道。     

Morphological and molecular analysis of genetic variability within isolates of <Emphasis Type="Italic">Corynespora cassiicola</Emphasis> from different hosts

Twenty-two isolates of Corynespora cassiicola obtained from cucumber, papaya, eggplant, tomato, bean, Vigna, sesame and Hevea rubber (Hevea brasiliensis) were analysed by morphological features, the differences of the ribosomal DNA internal transcribed spacer (rDNA-ITS) region sequence and the inter simple sequence repeat (ISSR) technique. Variability of morphological features was observed among the isolates. Pathogenicity tests showed that isolates from different hosts attacked Hevea rubber. Sequences of two outgroup taxa, C. proliferata and C. citricola, were downloaded from GenBank. The phylogenetic trees were constructed by using the rDNA-ITS region sequences from 24 Corynespora spp. isolates. In this analysis, the 24 sequences grouped into two clusters (A and B). Cluster A consists of sequences from all isolates of C. cassiicola; whereas cluster B consists of the two outgroup taxa, C. proliferata and C. citricola. However, the ITS region is conservative, and is not fit for studying differences among isolates. A total of 114 DNA fragments was amplified with 16 ISSR primers, among which 102 were polymorphic (89.5%). A dendrogram was created by the unweighted pair-group method with arithmetic averaging (UPGMA) analysis, and 22 isolates grouped into three clusters (C, D and E). Cluster C is composed of all of the Hevea rubber isolates, whereas cluster D is composed of nine isolates: four from papaya, five from cucumber, eggplant, bean, vigna and sesame. Cluster E is composed of two isolates from cucumber and tomato. These analyses showed that the genetic diversity was very rich among the tested isolates. There are no correlations between the morphological characteristics or rDNA-ITS region sequences of the 22 isolates and their host or geographical origin, but there is a link between ISSR clusters and their host origins. ISSR markers appear to be useful for intra-species population study in C. cassiicola.     

A controlled environment test for resistance to Soil-borne cereal mosaic virus (SBCMV) and its use to determine the mode of inheritance of resistance in wheat cv. Cadenza and for screening Triticum monococcum genotypes for sources of SBCMV resistance

Several wheat genotypes, including eight with known field responses, were evaluated for their reaction to Soil-borne cereal mosaic virus (SBCMV, genus Furovirus) by growing in naturally infested soil under controlled environment conditions. Virus antigen titres in the foliage 8–9 weeks after sowing mostly reflected the field responses, showing that growth chamber-based tests can be used to improve the speed and reliability of germplasm screening. Such tests were used to determine the mode of inheritance of the SBCMV resistance in cv. Cadenza, commonly used in UK wheat-breeding programmes. One hundred and eleven doubled haploid (DH) lines derived from an F ₁ of a cross between cvs Cadenza (resistant) and Avalon (susceptible) were evaluated. This DH population segregated for the reaction to SBCMV in a ratio of 1 : 1 (resistant : susceptible). This suggests that the SBCMV resistance is controlled by a single gene locus. As a first step towards identification of new sources of improved SBCMV resistance (e.g. immunity) as well as sources of the resistance to the virus vector, Polymyxa graminis , a set of 26 Triticum monococcum lines of diverse geographical origin was also screened. Most lines were susceptible to SBCMV, but one line of Bulgarian origin was resistant to the virus and possibly partially resistant to the virus vector.     

Brazilian Potato virus Y isolates identified as members of a new clade facilitate the reconstruction of evolutionary traits within this species

Potato virus Y (PVY) is a plant virus distributed worldwide that causes damage to several species of the Solanaceae family. It was established long ago that groups of PVY isolates defined by phylogenetic analyses correlate strongly with those demarcated by differential biological properties. Consequently, life‐history traits of this viral species can be inferred by phylogenetic analysis. In this study, characteristics of PVY isolates sampled in different tobacco fields in Brazil were analysed and most of the tested Brazilian PVY isolates were assigned to the recently described unconventional serogroup Y^U. The analysis of molecular diversity of the coat protein (CP) cistron from some Y^U isolates made it possible (i) to identify specific amino acid residues in the N‐terminal of the CP protein and (ii) to assign some Y^U isolates to a new PVY clade. The symptoms caused by isolates belonging to this new PVY ‘Brazilian’ clade and their ability to infect selected susceptible hosts led to the conclusion that neither veinal necrosis symptoms expressed on infected tobacco plants nor adaptation to potato or pepper hosts are ancestral characteristics of PVY. These observations suggest that PVY has gained a remarkable new biological property and broadened its host range over time.     

Influence of sowing density and spatial pattern of spring wheat (Triticum aestivum) on the suppression of different weed species

To better understand the potential for improving weed management in cereal crops with increased crop density and spatial uniformity, we conducted field experiments over two years with spring wheat ( Triticum aestivum ) and four weed species: lambsquarters ( Chenopodium album ) , Italian ryegrass ( Lolium multiflorum ), white mustard ( Sinapis alba ), and chickweed ( Stellaria media ). The crops were sown at three densities (204, 449, and 721 seeds m⁻²) and in two spatial patterns (normal rows and a highly uniform pattern), and the weeds were sown in a random pattern at a high density. In most cases, the sown weeds dominated the weed community but, in other cases, naturally occurring weeds were also important. There were strong and significant effects regarding the weed species sown, the crop density, and the spatial distribution on the weed biomass in both years. The weed biomass decreased with increased crop density in 29 out of 30 cases. On average, the weed biomass was lower and the grain yield was higher in the uniform compared to the row pattern in both 2001 and 2002. Despite the differences in weed biomass, the responses of L. multiflorum , S. media , and C. album populations to crop density and spatial uniformity were very similar, as were their effects on the grain yield. Sinapis alba was by far the strongest competitor and it responded somewhat differently. Our results suggest that a combination of increased crop density and a more uniform spatial pattern can contribute to a reduction in weed biomass and yield loss, but the effects are smaller if the weeds are taller than the crop when crop–weed competition becomes intense.     

Variation in the Fusarium section Liseola: pathogenicity and genetic studies of isolates of Fusarium moniliforme Sheldon from different hosts in Ghana

Fusarium moniliforme , the imperfect stage of the ascomycete Gibberella fujikuroi , is an economically important pathogen with a very wide host range. The genetic characteristics of isolates of the fungus collected from different regions of Ghana from maize, rice and sorghum were determined using the random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) techniques. The pathogenicity of the isolates was also compared on maize and rice. DNA fingerprints detected as RFLPs of ribosomal DNA and RAPDs separated the isolates into discrete groups which were generally host-related. The possibility of a sub-structuring of the maize population of the fungus into tissue-related subgroups was suggested by the results. A dendrogram of the relatedness of the isolates is presented. However, the pathogenicity of the isolates on rice, measured by their ability to cause 'bakanae' symptoms, did not resolve the isolates into the clearly defined groups suggested by the genetic studies, and maize isolates of the fungus could cause 'bakanae' symptoms to the same extent as rice isolates. Similarly, some isolates identified as rice-type isolates caused as much shoot stunting in maize as maize isolates. However, the effects of the isolates on root growth of maize seedlings showed a broad correlation with the defined genetic groups, with maize isolates of the fungus showing the greatest tendency to cause root stunting     

Genetic analyses of Pseudomonas syringae isolates from Belgian fruit orchards reveal genetic variability and isolate-host relationships within the pathovar syringae, and help identify both races of the pathovar morsprunorum

A collection of Pseudomonas syringae and viridiflava isolates was established between 1993 and 2002 from diseased organs sampled from 36 pear, plum and cherry orchards in Belgium. Among the 356 isolates investigated in this study, phytotoxin, siderophore and classical microbiology tests, as well as the genetical methods REP-, ERIC- and BOX- (collectively, rep-) and IS50-PCR, enabled identification to be made of 280 isolates as P. syringae pv. syringae (Pss), 41 isolates as P. syringae pv. morsprunorum (Psm) race 1, 12 isolates as Psm race 2, three isolates as P. viridiflava and 20 isolates as unclassified P. syringae. The rep-PCR methods, particularly BOX-PCR, proved to be useful for identifying the Psm race 1 and Psm race 2 isolates. The latter race was frequent on sour cherry in Belgium. Combined genetic results confirmed homogeneities in the pvs avii, and morsprunorum race 1 and race 2 and high diversity in the pv. syringae. In the pv. syringae, homogeneous genetic groups consistently found on the same hosts (pear, cherry or plum) were observed. Pathogenicity on lilac was sometimes variable among Pss isolates from the same genetic group; also, some Psm race 2 and unclassified P. syringae isolates were pathogenic to lilac. In the BOX analyses, four patterns included 100% of the toxic lipodepsipeptide (TLP)-producing Pss isolates pathogenic to lilac. Many TLP-producing Pss isolates non-pathogenic to lilac and the TLP-non-producing Pss isolates were classified differently. Pseudomonas syringae isolates that differed from known fruit pathogens were observed in pear, sour cherry and plum orchards in Belgium.