Intrapathotype diversity for aggressiveness and pathogen evolution in cultivar mixtures

ABSTRACT A model was developed and used to study the consequences of diversity for aggressiveness within pathotypes on pathogen evolution in two-component and four-component cultivar mixtures. It was assumed that, within a pathotype, a proportion of the isolates would have higher or lower spore efficacy than the average on a given host genetic background. Two situations were examined in which the pathogen can have either independent or negatively correlated values for spore efficacy on different cultivars. In the latter case, a pathogen genotype more aggressive than the average on a host genotype was always less aggressive on other host genotypes. In the simulations, isolates with greater aggressiveness relative to a host genotype were selected for and increased in frequency. However, because simple pathotypes always reproduced on the same host genotype whereas complex pathotypes were able to grow on several hosts, selection was faster for simple pathotypes. Pathotypes with two different levels of diversity for aggressiveness were compared with nondiversified pathotypes. In order to make comparisons, the effect of a 5 and 10% cost of virulence on the development of complex pathotypes was simulated. In general, increased diversity within pathotypes reduced the rate of increase of complex pathotypes in host mixtures, and this effect was stronger with greater frequencies of autodeposition of pathogen spores.     

Diversity,spatial variation,and temporal dynamics of virulences in the German leaf rust (<Emphasis Type="Italic">Puccinia recondita</Emphasis> f. sp. <Emphasis Type="Italic">secalis</Emphasis>) population in winter rye

A large collection of German rye leaf rust isolates was analysed to characterize the diversity, spatial variation and temporal dynamics of virulences. Virulence-avirulence phenotypes (=pathotypes) were determined on 23 host differentials. We found 93 pathotypes among 177 single-uredinial isolates in 2000, 201 pathotypes among 437 isolates in 2001, and 125 pathotypes among 213 isolates in 2002. In total, the 827 analyzed isolates represented 317 pathotypes. Frequency of virulences on the individual differentials varied from 2% to 97%. Eight of the differentials showed a high resistance level with virulence frequencies <10%. Virulence complexity of the isolates ranged from 3 to 21 with a mean of nine. The percentages of highly virulent isolates (>14 virulences) increased from 4 to 15% during the sampling period. A high level of virulence diversity was observed within and between individual sampling sites with Simpson indices around 0.9. Evenness indices ranged from 0.88 to 0.92. Four of the five most frequent pathotypes were found in each year but their frequency never exceeded 10%. Isolates with unusual virulence combinations could be clearly separated by principal component analysis. Location-specific pathotype frequencies were revealed in each year, but the frequency patterns varied across years. On four fields a considerable increase of highly virulent pathotypes occurred within 6 weeks during the epidemic. The high diversity of pathotypes as well as the fast accumulation of highly virulent pathotypes favour the adaptation of the pathogen to race-specific host resistances. More durable resistance might be achievable by combining new effective race-specific resistances with adult-plant and/or race-non-specific quantitative resistances.     

Selection for increased spore efficacy by host genetic background in a wheat powdery mildew population

ABSTRACT A field experiment was designed to test the hypothesis of for increased reproductive ability on different host genetic backgrounds within a wheat powdery mildew population. Studies have suggested that, in host mixtures, such selection could increase the reproduction rate of simple patho-types that always develop on the same host genetic background, whereas complex pathotypes should not be affected because they infect different host genotypes. In our experiment, the Erysiphe graminis population reproduced for successive generations on cvs. Orkis and Etecho, either grown as pure stands or in a mixture. In an additional treatment, the host cultivar changed after each generation. Isolates were sampled in April and, after seven pathogen generations, in July. At the second sampling date and for pure stands only, mean spore efficacy was greater on the host from which isolates were sampled than on the other one. This was attributed to selection within the pathogen population for better spore efficacy on the host genetic background. This selection was independent of the virulence genes carried by the isolates. The possibility of a phenotypic plasticity effect was tested and rejected.     

Effect of wheat cultivar mixtures on populations of Puccinia striiformis races*

This study quantifies the frequency of simple and complex races (races that can infect two or more components) of Puccinia striiformis in mixtures of wheat cultivars possessing different race-specific resistance genes. Treatments were designed so that the complex race changed depending on the host mixture, thus enabling us to observe the influence of pathogen complexity in different genetic backgrounds. Six cultivar mixtures and one pure stand of winter wheat were inoculated with three races of P. striiformis at two locations for two seasons. Potted plants of three winter wheat cultivars, each susceptible to one of the three races of the pathogen, were used to sample the pathogen during the field epidemics. Disease incidence on the differential cultivars was used to calculate the proportion of the three races in each treatment. The specific cultivars included in the mixtures influenced the frequencies of the three races. Increasing the number of virulent races in a mixture reduced the frequency of the complex race relative to the other two races. The results suggest that genetic background of the pathogen race, host composition, and interaction among pathogen races may be as important as cost of virulence in determining race frequencies in mixtures.     

Utility of defeated resistance genes to powdery mildew,Erysiphe graminis f. sp. hordei,in spring barley variety mixtures

In 1980 and 1981 experiments have been performed to study the utility of defeated resistance genes to powdery mildew in spring barley variety mixtures. For this purpose the epidemic development of powdery mildew in pure stands and mixtures of four spring barley varieties was monitored. In three varieties the resistance was overcome several years ago. One variety is still resistant. Changes in frequencies of corresponding virulence genes were also studied.It was found that the variety mixtures slowed down the mildew epidemic only at the beginning of the season. Some reduction was lost later on since the pure stands reached their saturation level of infection at an earlier date than the more healthy mixtures. The level of reduction of infection depended on the composition of the mixtures and the number of components. Two-way mixtures appeared to be very risky due to the high frequency of corresponding combinations of virulence genes in the pathogen. Furthermore, two-way mixtures appeared to be unable to reduce the infection rate sufficiently when the infection pressure was high.In 1981 the frequencies of combinations of corresponding virulence genes increased very much in the Diva-Mazurka mixture but declined in the pure stands of both varieties.On the basis of these results it is concluded that the utility of defeated resistance genes involved in this study is very limited.The low number and the irregular distribution of the Dutch spring barley varieties over the different resistance groups prohibit the use of variety mixtures in the Netherlands.Samenvatting In 1980 en 1981 is onderzoek verricht over de bruikbaarheid van doorbroken resistentiegenen tegen meeldauw,Erysiphe graminis f. sp.hordei, in zomergerstrassenmengsels. Daartoe werd de epidemieontwikkeling in monocultures en mengsels van vier zomergerstrassen bestudeerd. Van drie rassen is de resistentie reeds verscheidene jaren doorbroken. Eén ras is nog resistent. De frequenties van compatibele meeldauwvirulentiegenen werden eveneens bepaald.Het bleek dat in vergelijking met de monocultures in de rassenmengsels de meeldauwepidemie alleen in het begin van het seizoen werd afgeremd. Later in het seizoen ging weer enige reductie in aantasting van de mengsels verloren doordat in de monocultures de aantasting eerder het verzadigingspunt bereikte dan in de minder aangetaste mengsels. De mate waarin de aantasting gereduceerd werd was afhankelijk van de aard en het aantal componenten in het mengsel. Twee-componentenmengsels bleken vrij riskant te zijn vanwege de aanwezigheid van combinaties van corresponderende virulentiegenen in de meeldauwpopulatie. Verder bleken tweecomponentenmengsels niet in staat de infectiesnelheid afdoende af te remmen wanneer de infectiedruk hoog werd.De frequentie van combinaties van corresponderende virulentiegenen nam sterk toe in het Diva-Mazurka-mengsel doch nam af in de monocultures van deze beide rassen.Gezien de verkregen resultaten lijkt de bruikbaarheid van de in dit onderzoek betrokken doorbroken resistentiegenen erg beperkt. Het geringe aantal zomergerstrassen op de rassenlijst en de ongelijke verdeling over de te onderscheiden resistentiegroepen belemmeren de toepassing van rassenmengsels in de Nederlandse akkerbouw.     

Distribution of pathotypes with regard to host cultivars in French wheat leaf rust populations

ABSTRACT Isolates of wheat leaf rust collected from durum and bread wheat cultivars in France during 1999-2002 were analyzed for virulence on 18 Thatcher lines with single genes for leaf rust resistance (Lr genes). Sampling focused on the five most widely grown bread wheat cultivars (two susceptible and three resistant) to allow statistical comparison of diversity indexes between the cultivars. Leaf rust populations from durum and bread wheats were different. The diversity of the bread wheat leaf rust pathotypes, as measured by the Shannon index, ranged from 2.43 to 2.76 over the 4 years. Diversity for wheat leaf rust resistance was limited in the host since we postulated only seven seedling resistance genes in the 35 cultivars most widely grown during 1999-2002. Leaf rust populations were strongly differentiated for virulence within bread wheat cultivars, and diversity was higher on those that were resistant, mainly due to a more even distribution of virulence phenotypes than on susceptible cultivars. The pathogen population on the susceptible cv. Soissons was largely dominated by a single pathotype (073100), whereas all other pathotypes virulent on cv. Soissons either decreased in frequency or remained at a low frequency during the period studied. Several pathotypes including the most complex one were found only on resistant cultivars, even though most of them were virulent on the susceptible cv. Soissons. Specific interactions were necessary, but not always sufficient, to account for pathotype distribution and frequencies on the cultivars, suggesting that selection for virulence to host resistance genes is balanced by other selective forces including selection for aggressiveness.     

Selection of pathotypes of Erysiphe graminis f.sp. hordei in pure and mixed stands of spring barley

Pathotypes of Erysiphe graminis f.sp. hordei were monitored at fortnightly intervals in pure and mixed stands of spring barley during the course of mildew epidemics in two field trials. Mixtures were composed of cultivars with Arabische (gene Mla12 ), Laevigatum ( Ml(La) ), and Monte Cristo ( Mla9 ) resistance, respectively. The three-way mixtures were either random or, in 1989, laid out as one-row mixtures (i.e., regularly alternating rows of different genotypes) or three-row mixtures (i.e., regularly alternating three-row strips of different genotypes), respectively. In 1990 only random mixtures and six-row mixtures were compared with pure stands. The virulence complexity (i.e., the average number of virulence factors per isolate with reference to Mla12, Ml(La) , and Mla9 ) was always maximal in the random mixtures. In 1989, linear regression of complexity on mildew generations gave significant b -values (slopes) of 0·049, 0·031, and 0·025 in the random mixture, one-row mixture, and three-row mixture, respectively; the b -value from pure stands was not significant. In 1990, another sampling technique allowed selection to be observed on each genotype in the mixtures separately. In the random mixture b -values were 0·048, 0·064 and 0·017 (not significant) on Mla12, Ml(La) , and Mla9 cultivars respectively. In six-row mixtures and in pure stands, there was no significant increase in complexity ( b > 0) on any of the mixture components. Although the frequency and relative fitness of complex pathotypes were higher in all types of mixtures than in pure stands, selection towards complex races was much less intense in row mixtures than in random mixtures in both field trials.     

Resistance to Turnip mosaic virus in Brassica rapa and B. napus and the analysis of genetic inheritance in selected lines

Forty-two Brassica rapa and Brassica napus lines were tested for resistance to Turnip mosaic virus (TuMV) isolates representing the three major pathotypes in Europe. Of these lines, 11 were susceptible to all pathotypes; nine were resistant to one pathotype; eight were resistant to two pathotypes; and 14 were resistant to all three pathotypes. Of the lines tested, 23 were either able to, or had the potential to, discriminate between two different pathotype-3 isolates. Genetic models for inheritance of resistance were proposed for four B. rapa lines: Jong Bai No. 2 had dominant resistance to pathotype 1 conferred by a single allele; PI418957C and Jin G 55 had recessive resistance to pathotype 4 where a single allele was required; PI418957C also had recessive resistance to pathotype 3 where a model with one of two epistatic, unlinked loci was proposed. Jong Bai No. 1 also had recessive resistance to pathotype 3, apparently conferred by alleles at three loci, where any two of the three loci were epistatic and required for resistance.     

Evolution of a pathogen population in host mixtures: simple race–complex race competition

The evolution of the genetic structure of a pathogen population was studied in a varietal mixture with an epidemic simulator based on the model EPIMUL. The pathogen population was composed of simple races able to develop on only one component of the mixture and a complex race which developed on all mixture components. The effects on the simple race–complex race competition of a cost of virulence, of density dependence and of differential adaptation were studied. The selection for simple or complex races in the pathogen population did not depend on initial race frequencies. For a given multiplication rate, complex race frequency increased faster when the spore dispersal gradient was shallow, when distribution of initial disease was generalized, when amount of initial disease was reduced and when the number of mixture components was increased. This was attributed to a better efficacy of the mixture in controlling simple races, resulting in a higher relative fitness of the complex race. For measured values of density dependence or differential adaptation effects, the complex race was at a higher frequency after a mean number of pathogen cycles between 2.5 and 5. The effect of the cost of virulence was stronger and, in certain situations, could result in selection for simple races. In the conditions of our simulations and with the effects tested, stabilization of the pathogen population in host mixtures was unlikely to occur. However, more information is needed concerning the rate at which complex races could evolve and how quickly mixture resistance could be eroded.     

Effects of Wheat Cultivar Mixtures on Epidemic Progression of Septoria Tritici Blotch and Pathogenicity of Mycosphaerella graminicola

ABSTRACT The effects of host genotype mixtures on disease progression and pathogen evolution are not well understood in pathosystems that vary quantitatively for resistance and pathogenicity. We used four mixtures of moderately resistant and susceptible winter wheat cultivars naturally inoculated with Mycosphaerella graminicola to investigate impacts on disease progression in the field, and effects on pathogenicity as assayed by testing isolate populations sampled from the field on greenhousegrown seedlings. Over 3 years, there was a correspondence between the mixtures' disease response and the pathogenicity of isolates sampled from them. In 1998, with a severe epidemic, mixtures were 9.4% less diseased than were their component pure stands (P = 0.0045), and pathogen populations from mixtures caused 27% less disease (P = 0.085) in greenhouse assays than did populations from component pure stands. In 1999, the epidemic was mild, mixtures did not reduce disease severity (P = 0.39), and pathogen populations from mixtures and pure stands did not differ in pathogenicity (P = 0.42). In 2000, epidemic intensity was intermediate, mixture plots were 15.2% more diseased than the mean of component pure stands (P = 0.053), and populations from two of four mixtures were 152 and 156% more pathogenic than the mean of populations from component pure stands (P = 0.043 and 0.059, respectively). Mixture yields were on average 2.4 and 6.2% higher than mean component pure-stand yields in 1999 and 2000, respectively, but the differences were not statistically significant. The ability of mixtures challenged with M. graminicola to suppress disease appears to be inconsistent. In this system, host genotype mixtures evidently do not consistently confer either fitness benefits or liabilities on pathogen populations.     

Seasonal changes in pathotype complexity in French populations of barley powdery mildew

Airborne conidia of Erysiphe graminis f.sp. hordei were sampled in three regions and a single locality in the northern part of France for 2 years. Sampling was carried out in early spring, in late spring and in autumn, in order to separate the effects of winter barley cultivars, carrying few specific resistance alleles, and of spring barley cultivars, carrying diverse resistance alleles, on the structure of the pathogen population. Although complex pathotypes with three to 10 virulences were selected by spring cultivars, simple pathotypes, including a pathotype with the single unnecessary virulence allele Va22 , which formed a clear majority of the samples, remained dominant in early spring, when winter but not spring cultivars were growing. In early spring, simple pathotypes were more prevalent in the north, where the winter cultivars represented 90% of the barley acreage, than in the east, where winter cultivars represented 65%. In the west, the frequency of simple pathotypes was limited compared to the north, possibly because of the resistance allele Mlg in winter cultivars. The high frequency of simple pathotypes in early spring could be explained by a differential adaptation between simple and complex pathotypes or by delayed epidemics on spring cultivars compared to winter cultivars.     

Host frequency and density effects on powdery mildew and yield in mixtures of barley cultivars

The effects of frequency and density of susceptible plants on barley powdery mildew epidemics were studied in a combined set of addition and replacement series of field trials. In the addition series, plant densities in pure stands of three cultivars, Rambo, Rodos and Grosso (susceptible, moderately resistant and immune, respectively) were varied six-fold. In the replacement series, the three possible two-way mixtures were analysed at different frequencies but at a density corresponding to the maximum pure stand density. Disease and yield were assessed on a per-plant basis. In the pure stands, tillering reduced the range of densities from six-fold to between three- and four-fold, while in the mixtures, frequencies changed only slightly over time, indicating that competitive interactions among the cultivars were roughly equal. Yield per plant decreased logarithmically with increasing density as expected. However, yield per seed head was not correlated with the final number of heads per plot, indicating low competition among heads even at the highest density. Disease in susceptible pure stands increased strongly with decreasing density in 1994 and to a lesser degree in 1995. These differences could have been caused by differences in plant nutritional status and consequent epidemiological effects. Disease reduction on the susceptible cultivars in mixtures varied between 33% and 71% among years. Depending on the length and strength of the epidemic, the effects of host density and frequency on disease severity varied substantially among years.     

2013-2016年甘肃省小麦条锈菌生理小种变异监测

2013-2016年对1 000份甘肃省小麦条锈菌标样进行生理小种鉴定和监测, 结果表明:共监测到56个生理小种及致病类型。贵农22致病类群已占据优势地位, 出现频率由2013年的28.72%上升到2016年的66.76%。新小种条中34号(原代号贵22-9)出现频率由2013年的11.28%上升到2016年的34.85%, 居第一位; 条中32号出现频率由2013年的15.38%下降到2016年的9.12%, 居第二位; 贵22-14致病类型2016年出现频率6.43%, 居第三位; 条中33号出现频率由13.85%降为5.63%, 居第四位。次要小种HY4、HY8、水11-4、水11-5、水11-7及贵22-8、贵22-45、贵22-74等类型出现频率较低, 在051%~615%之间。毒性频率分析发现, 到2016年对 Yr9、Yr3b + Yr4b、YrSu、Yr26 基因的毒性频率均超过58%, 其中对 Yr26 超过72%, 甘肃省小麦条锈菌群体已进入以条中34号为主、贵农22致病类群占优势的新的变异活跃期。抗锈育种应以针对条中34号、条中32号、条中33号为主, 兼顾贵农22、Hybrid 46和水源11致病类群中其他类型。     

Pepper resistance-breaking tobamoviruses: Can they co-exist in single pepper plants?

We previously reported a procedure for identifying pathotypes of the tobamoviruses infecting the L-resistant genotypes of pepper based on the polymerase chain reaction (PCR) (Tenllado et al., 1994). We have now used this method to assess pathotype incidence in virus isolates from field pepper samples representing three successive epidemic episodes in southeastern Spain, and to analyse the interaction of pathotypes during experimental infections in pepper plants. The majority of virus isolates corresponded to the P_1,2 pathotype, and only three out of twenty behaved as P_1,2,3 pathotypes. Interestingly, restriction enzyme analysis of the PCR-amplified products distinguished two restriction subgroups on each P_1,2 and P_1,2,3 pathotype, referred to as restrictopatterns I₁ and I₂ or II₁ and II₂, respectively. Experimental coinoculations of pepper plants with mixtures of two different pathotypes showed coexistence between them when inoculations proceeded simultaneously. However, reciprocal cross-protection was observed between P_1,2 and P_1,2,3 pathotypes when they were successively inoculated, while no cross-protection was observed between P₁ and either P_1,2 or P_1,2,3 pathotypes. The potential of the PCR-based method for detecting heterogeneity within P_1,2 and P_1,2,3 pathotypes, and the possibility of genetically engineered resistance to those viruses by genetic transformation with viral coat protein genes are briefly discussed.     

Pathotypic diversity of Hyaloperonospora brassicae collected from Brassica oleracea

Downy mildew caused by Hyaloperonospora brassicae is an economically destructive disease of brassica crops in many growing regions throughout the world. Specialised pathogenicity of downy mildews from different Brassica species and closely related ornamental or wild relatives has been described from host range studies. Pathotypic variation amongst Hyaloperonospora brassicae isolates from Brassica oleracea has also been described; however, a standard set of B. oleracea lines that could enable reproducible classification of H. brassicae pathotypes was poorly developed. For this purpose, we examined the use of eight genetically refined host lines derived from our previous collaborative work on downy mildew resistance as a differential set to characterise pathotypes in the European population of H. brassicae. Interaction phenotypes for each combination of isolate and host line were assessed following drop inoculation of cotyledons and a spectrum of seven phenotypes was observed based on the level of sporulation on cotyledons and visible host responses. Two host lines were resistant or moderately resistant to the entire collection of isolates, and another was universally susceptible. Five lines showed differential responses to the H. brassicae isolates. A minimum of six pathotypes and five major effect resistance genes are proposed to explain all of the observed interaction phenotypes. The B. oleracea lines from this study can be useful for monitoring pathotype frequencies in H. brassicae populations in the same or other vegetable growing regions, and to assess the potential durability of disease control from different combinations of the predicted downy mildew resistance genes.     

Is virulence phenotype evolution driven exclusively by Lr gene deployment in French Puccinia triticina populations?

Puccinia triticina is a highly damaging wheat pathogen. The efficacy of leaf rust control by genetic resistance is mitigated by the adaptive capacity of the pathogen, expressed as changes in its virulence combinations (pathotypes). An extensive P. triticina population survey has been carried out in France over the last 30 years, describing the evolutionary dynamics of this pathogen in response to cultivar deployment. We analysed the data set for the 2006–2016 period to determine the relationship between the Lr genes in the cultivars and virulence in the pathotypes. Rust populations were dominated by a small number of pathotypes, with variations in most of the virulence frequencies related to the corresponding Lr gene frequencies in the cultivated landscape. Furthermore, the emergence and spread of a new virulence matched the introduction and use of the corresponding Lr gene (Lr28), confirming that the deployment of qualitative resistance genes is an essential driver of evolution in P. triticina populations. However, principal component analysis (PCA) revealed that certain pathotype–cultivar associations cannot be explained solely by the distribution of Lr genes in the landscape. This conclusion is supported by the predominance of a few pathotypes on some cultivars, with the persistence of several other compatible pathotypes at low frequencies. Specific interactions are not, therefore, sufficient to explain the distribution of virulence in rust populations. The hypothesis that quantitative interactions between P. triticina populations and bread wheat cultivars—based on differences in aggressiveness—is also a driver of changes in pathotype frequencies deserves further investigation.     

云南省水稻细菌性条斑病菌的致病型划分和水稻抗性资源的鉴定

 为明确云南省水稻细菌性条斑病菌(Xanthomonas oryzae pv. oryzicola, Xoc)的致病力分化以及不同类型水稻品种对Xoc的抗感特性,通过针刺接种法将云南省8个稻区采集的86株Xoc菌株,接种于6个携带不同抗性基因的水稻鉴别品种(IRBB4、IRBB5、IRBB14、IRBB18、IRBB21和IR24)。根据这些菌株在鉴别品种上的毒力差异进行了UPGMA聚类分析,将其划分为9个致病型(Ⅰ型 ~ Ⅸ 型)。其中,Ⅰ型为优势菌群,分布频率为50.5%。对不同稻区的优势菌群进行分析,发现云南省各稻区Xoc的致病型呈多样性分布,以强毒力的Ⅰ型为高频率致病型。选用 Ⅰ 型、Ⅱ 型和 Ⅵ 型代表菌株对云南省的80个主栽和区试水稻品种进行抗性评价,对3个致病型表现抗性的材料比例分别为30.0%、35.0%和57.5%。筛选出9个对3种致病型都表现为抗性的品种,其中“Deyou16”和“Changgui2”表现为高抗。研究结果可为云南省防治水稻细菌性条斑病的水稻区域性布局和抗性品种的利用提供理论依据。     

AAL-Toxin-Deficient Mutants of Alternaria alternata Tomato Pathotype by Restriction Enzyme-Mediated Integration

ABSTRACT Host-specific toxins are produced by three pathotypes of Alternaria alternata: AM-toxin, which affects apple; AK-toxin, which affects Japanese pear; and AAL-toxin, which affects tomato. Each toxin has a role in pathogenesis. To facilitate molecular genetic analysis of toxin production, isolation of toxin-deficient mutants utilizing ectopic integration of plasmid DNA has been attempted. However, the transformation frequency was low, and integration events in most transformants were complicated. Addition of a restriction enzyme during transformation has been reported to increase transformation frequencies significantly and results in simple plasmid integration events. We have, therefore, optimized this technique, known as restriction enzyme-mediated integration (REMI), for A. alternata pathotypes. Plasmid pAN7-1, conferring resistance to hygromycin B, with no detectable homology to the fungal genome was used as the transforming DNA. Among the three restriction enzymes examined, HindIII was most effective, as it increased transformation frequency two-to 10-fold depending on the pathotype, facilitating generation of several hundred transformants with a 1-day protocol. BamHI and XbaI had no significant effect on transformation frequencies in A. alternata pathotypes. Furthermore, the transforming plasmid tended to integrate as a single copy at single sites in the genome, compared with trials without addition of enzyme. Libraries of plasmid-tagged transformants obtained with and without addition of restriction enzyme were constructed for the tomato pathotype of A. alternata and were screened for toxin production. Three AAL-toxin-deficient mutants were isolated from a library of transformants obtained with addition of enzyme. These mutants did not cause symptoms on susceptible tomato, indicating that the toxin is required for pathogenicity of the fungus. Characterization of the plasmid integration sites and rescue of flanking sequences are in progress.     

Pathotypic variation in turnip mosaic virus with special reference to European isolates

A collection of 124 isolates of turnip mosaic virus was gathered from around the world, principally from European countries, and characterized by inoculation to four differential lines of Brassica napus (oilseed rape and swede). Three symptom phenotypes were induced—apparent immunity, local infection only, or systemic infection. Twelve distinct patterns, i.e. pathotypes, were observed. Three pathotypes were predominant in the collection; pathotype 1 isolates, which were the most common, did not overcome any of the most extreme sources of resistance in the differential lines. Of the other two, pathotype 3 isolates overcame one of the major sources of resistance and pathotype 4 isolates overcame all sources of resistance. The distribution of pathotypes within Europe was examined. No pathotype was confined to any geographical area, although pathotype 4 isolates were not found in southern Europe or Asia. Most isolates (90) originated from Brassica hosts, while others were from other cruciferae genera (19) or non-crucifers (5). The species of plant that the isolates originated from was not clearly related to the pathotype of the isolates. Resistance to pathotype 1 isolates is controlled by a dominant allele in one of the differential lines, and resistance sources are being examined in the other lines. Isolates belonging to pathotype 1 appeared to be able to mutate readily to overcome the resistance in one of the rape differential lines, but no isolates appeared to mutate to overcome the other major source of resistance in the differentials. The implications of the results for disease control strategies are discussed.     

华南水稻白叶枯病菌致病性分化检测与分析

为了解华南稻区水稻白叶枯病菌的致病性分化和变异动态,采集华南地区水稻白叶枯病病叶标样分离病原菌,应用中国鉴别寄主IR26、南粳15、爪哇14、特特普、金刚30和国际水稻已知抗病基因的近等基因系IRBB5、IRBB13、IRBB3、IRBB14、IRBB2、IR24两套鉴别寄主,在水稻孕穗期采用剪叶法接种,依据寄主和菌株的互作反应检测病菌的致病性分化。结果显示,参试菌株可划分为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅸ六个致病型和R1、R2、R3、R4、R5、R8、R10七个致病小种。Ⅴ、Ⅳ致病型和R8、R5小种出现频率分别为27.40%、19.30%和44.67%、15.34%,为华南稻区优势种群。Ⅸ、Ⅴ、Ⅳ致病型和R8、R5小种对500份华南稻区品种资源的致病率依次为96.40%、95.00%、50.40%、62.00%和42.60%;Ⅸ致病型毒性最强且发展很快;强致病菌系Ⅴ型已替代Ⅳ型发展为华南优势致病菌系。