Effect of interrupted leaf wetness periods on pustule development of Puccinia recondita f. sp. tritici on wheat

In growth chamber experiments, seedlings of a susceptible wheat cultivar (Rubis) were inoculated with urediospores of wheat leaf rust. Inoculated seedlings were incubated in a moist chamber. The period of exposure to leaf wetness varied in duration and progressed with or without one or more interruptions of variable duration. During the interruptions the leaves with germlings were dry. At near-optimal temperature, leaf wetness periods of six h resulted in 60 to 65% of the pustules produced with 12 or 24 h wetness periods. Interruption of a 6 h leaf wetness period by a 1 h dry period was most damaging to the rust germlings at about three to four h after inoculation. The damage became visible as a reduction in the number of resulting pustules and as a prolongation of the median latency period. Epidemiological implications of leaf wetness interruptions are discussed.     

The significance of VA-Mycorrhiza to future arable farming in the Netherlands

Present conditions of high inputs and narrow rotations render the importance of vesicular-arbuscular mycorrhiza (VAM) in arable farming in the Netherlands rather low. Changes expected in the near future under the influence of economic pressures and environmental problems may change this situation. This is discussed in the light of some such expected changes in Dutch arable farming.De huidige Nederlandse akkerbouw is zeer intensief. Ze heeft in het algemeen een nauwe rotatie met slechts enkele gewassen (aardappelen, granen en suikerbieten) en een ruime toepassing van kunstmest en bestrijdingsmiddelen. Het is te verwachten dat er onder invloed van toenemende problemen met het milieu, afnemende bodemproduktiviteit en lagere prijzen voor nu geteelde gewassen ten gevolge van overschotten een tendens zal zijn in de richting van een meer geïntegreerde landbouw. In de bestaande akkerbouw werken zowel hoge kunstmestgiften (met name N en P) en bestrijdingsmiddelen negatief op VAM-vorming. Naarmate het gebruik van kunstmest en bestrijdingsmiddelen in de nabije toekomst wellicht zal verminderen, kan worden verwacht dat zowel de aanwezigheid als het belang van VAM in de nederlandse landbouw zal toenemen. Het laatste zal vooral het geval zijn indien de vruchtwisseling wordt verruimd met de teelt van andere gewassen (zoals vlinderbloemigen) met een hogere VAM-afhankelijkheid dan die van het huidige gewasassortiment. Het zou zeer verstandig zijn zich reeds nu te beraden over vormen van bedrijfsvoering die VAM zoveel mogelijk stimuleren en daardoor meer profijt trekken van de door deze symbiose geboden voordelen.     

Shaping a focus: wind and stochasticity

The diffusion theory of focus development permits development of a class of related models of different complexity. The basis of the present study is an earlier model which simulates a dual spore dispersal mechanism, consisting of a short-distance and a long-distance mechanism. This model was further elaborated by adding stochasticity and an external directional force simulating wind. Stochasticity was simulated through randomization of lesions produced by spores dispersed by the long-distance mechanism. The model was run twelve times for three values of spore partitioning over the two dispersal mechanisms combined with four values of wind speed. Model responses were sampled at three simulation time values: 40, 90, and 100 days. A major model response was the shape of a single focus, studied through snap-shot pictures of disease severity over the field. Real life phenomena, non-circular mother or central focus with ragged front and daughter foci, were reproduced. Throughout the simulations, maximum disease severity remained at the location of the initial inoculation. However, after all available sites around the centre were exhausted, the geometrical centre of the focus was apparently moved downwind. Number of daughter foci and their relative size were strongly dependent on the spore partitioning over the two dispersal mechanisms.     

小麦叶锈菌在感病寄主上发育的组织病理学和超微结构研究

 应用荧光显微技术、微分干涉技术和生物电镜技术,系统地研究了小麦叶锈菌在感病寄主上的发育过程及其超微结构特征。小麦叶锈菌在感病品种上的发育过程可分为几个明显的阶段,即孢子的萌发、附着胞的形成、气孔下囊的分化、初生菌丝和次生菌丝的形成和生长、吸器母细胞和吸器的形成、夏孢子床和夏孢子堆的产生以及夏孢子的形成。小麦叶锈菌的胞间菌丝呈丝状,生长和分枝通常沿寄主细胞壁进行。胞间菌丝与寄主细胞的接触诱导了吸器母细胞的分化,吸器母细胞在与寄主细胞壁的接触部位发育形成入侵栓,穿透寄主细胞壁后于细胞内形成吸器。胞间菌丝和吸器母细胞均含有双核,而成熟吸器则含有单核。经常规染色后,胞间菌丝和吸器母细胞的壁与隔膜均可分辨出由多层构成。     

Growth of Xanthomonas campestris pv. campestris populations at constant and variable temperatures

Quantitative data were collected to describe the relation between temperature and growth of the cabbage black rot pathogen,Xanthomonas campestris pv.campestris (Xcc). Relative growth rates derived from experiments at constant temperatures were used in dynamic simulation of bacterial population development. The relative growth rates were adequate to simulate growth ofXcc populations at constant temperatures but overestimated growth of populations at variable temperatures. This finding gives rise to the hypothesis, that under field conditions, disease development is slower than is expected on the basis of growth parameters obtained from studies with constant temperatures.     

Systems approaches in epidemiology and plant disease management

This contribution to Zadoks’ liber amicorum reviews the developments in quantitative epidemiology during the last decades. It elucidates the progress in this field and shows how empirical crop protection with many phenomenological aspects transformed into a science-based (inter) discipline. The availability of experimental tools and the rapid development and introduction of computers enabled the application of systems approaches which stimulated a revolution in thinking and caused a considerable improvement of strategic and tactical decision making in crop protection. Zadoks played a crucial role in that development.     

Effect of culture filtrates of seventeen fungicolous fungi on sporulation of cucumber powdery mildew

Culture filtrates of 17 different fungal species thriving upon other fungi were tested for their ability to reduce sporulation of cucumber powdery mildew,Sphaerotheca fuliginea.All culture filtrates reduced the number of healthy conidiophores. However, the differences in activity between the various treatments were not as conspicuous as after application of spore suspensions. The best results were obtained with culture filtrates ofCalcarisporium arbuscula. These reduced the number of healthy conidiophores to ca. 2% of the unsprayed control plants.     

Molecular aspects of the potato — Phytophthora infestans interaction

The fungal pathogenPhytophthora infestans is the causal organism of late blight, one of the most devastating diseases of potato. In the past, various aspects of the potato-P. infestans interaction have been studied extensively. In this paper we briefly review the current knowledge of the molecular events associated with the interaction and in addition we discuss a new approach for analyzing the molecular basis of pathogenicity ofP. infestans.     

Effects of growth stage, leaf position and temperature on adult-plant resistance of wheat infected by Puccinia recondita f.sp. tritici

Growth stage, leaf position and temperature effects on components of resistance to Puccinia recondita f.sp. tritici in wheat ( Triticum aestivum ) were studied. Inoculation of Era, Glenlea, Sinton and RL6044 plants at five growth stages showed that latent period increased and uredinium numbers decreased as each adult-plant-resistant wheat matured. Growth stage had little effect on latent period in the susceptible cultivar Morocco. No significant differences in latent period determined on the flag, flag -1 and flag -2 leaves of the same plant were observed for Era, Glenlea and RL6044. However, leaf age should be considered in assessing adult-plant resistance because the latent period of P. recondita f.sp. tritici in flag leaves of Sinton was shorter than that determined for the flag-1 and flag -2 leaves. Fewer uredinia developed on the flag -2 leaf of Glenlea than on the flag leaf. The latent period of leaf rust in Era, Glenlea, RL6044 and Morocco was significantly longer at 15°C than at 21°C, but temperature did not affect Sinton to the same degree. Uredinium size was a sensitive criterion of adult-plant resistance. Uredinium development in line RL6044 was significantly restricted at 15°C, but uredinia on Era, Glenlea and Sinton at 15°C were as large as those at 21°C. On the susceptible Morocco uredinia were larger at 15°C than at 21°C.     

Surveys of cereal diseases and pests in the Netherlands. 6. Occurrence of insect pests in winter wheat

Between 1980 and 1986, commercial winter wheat fields at the milky-ripe stage were surveyed for insect pests. Leaf injuring insects were common. On average 42% and 95% of the surveyed fields were injured by leaf miners and the cereal leaf beetle, respectively. On average 14% of the leaves was skeletonised by the cereal leaf beetle and leaf injury tended to be high in seasons after winters with low temperatures. Blossom midges were the most important midges. In the period 1980–1984, the orange and lemon blossom midge infested 71 and 21% of the surveyed fields, respectively, Insecticides were mainly used to control aphids. Despite control, on average 80% of the fields and 22% of the tillers were infested by aphids.     

Genetic mapping and utilization of quantitative trichome-mediated insect resistance in potato

Introgression of trichome-mediated insect resistance from the wild speciesSolanum berthaultii has become a major focus of the potato improvement program at Cornell University during the past twelve years. Several quantitative characters are involved in this resistance which is effective against a wide range of pest types. Correlative biochemical assays have been developed to assay specific components of the resistance, and the effects of the resistance on the target pests have been studied. Quantitative laboratory assays and specific measurements of insect behavior and biology have increased the precision of selection and enable the investigation of the genetic control of the resistance.We are currently using restriction fragment length polymorphisms (RFLPs) for genetic mapping of factors controlling the trichome traits fromS. berthaultii. Backcrosses to both the wild and the cultivated species parents have been evaluated for phenotypes contributing to the resistance mechanism, including trichome density, sucrose ester and polyphenol oxidase production by the trichomes, and the enzymatic browning reaction responsible for insect entrapment. Genetic maps are being developed for these progenies, using RFLP markers previously mapped in potato. Field and greenhouse trials under insect infestations are also being conducted with the mapping progeny. Our goal is to locate genes responsible for quantitative insect resistance by correlating RFLP variation at mapped loci with the trichome phenotypes and insect resistance. Genetic markers for these traits will be useful in transfer of the effective wild chromosomal segments into and among tetraploid potatoes, and for a better understanding of the resistance mechanism.     

Towards the isolation of resistance genes by transposon targeting in potato

A general strategy for the isolation of disease resistance genes is presented, employing a two-step approach of transposon targeting near genes of interest followed by transposon tagging. A library of transposon (Ac/Ds) transformants in a self fertile potato diploid are being mapped by deriving genomic DNA probes flanking the transposon containing T-DNA insertions with the inverse polymerase chain reaction and using these probes for RFLP analysis. We have produced a large number of transposon (Ac/Ds) transformants in a self fertile potato diploid. Genomic DNA probes, flanking the transposon containing T-DNA insertions, are produced by the inverse polymerase chain reaction (IPCR) and mapped by restriction fragment length polymorphism (RFLP) analysis in a segragating potato location. A transposon mapped close to a resistance gene can be recombined cis to the gene and used for efficient transposon targeting due to preferential transposition to linked sites.     

Prospects for improving potato crop protection against aphid vectors

Conventional ways to control aphid vectors on potato crops rely essentially on cultural and chemical methods. The extensive use of insecticides, too often applied regardless of real needs and consequences, has resulted in more and more failures in both controlling aphids and virus spread. This paper stresses the need to introduce more rationale in the operation of existing methods by improving the efficiency of some of them like forecasting systems, to assess the potential of others like fertilization and semiochemicals and to put most efforts on resistant cultivars so that integrated pest management can be operated in the near future.     

Effects of imazalil on sterol composition of sensitive and DMI-resistant isolates of Penicillium italicum

Imazalil differentially inhibited dry weight increase of 10-hour-old germlings of wild-type and DMI-resistant isolates ofPenicillium italicum in liquid malt cultures. EC₅₀ values ranged from 0.005 to 0.27 g ml^–1. In all isolates ergosterol constituted the major sterol (over 95% of total sterols) in the absence of the fungicide. Therefore, DMI-resistance cannot be associated to a deficiency of the C-14 demethylation enzyme in the ergosterol biosynthetic pathway. Imazalil treatment at concentrations around EC₅₀ values for inhibition of mycelial growth resulted in a decrease in ergosterol content and a simultaneous increase in 24-methylene-24,25-dihydrolanosterol content in all isolates. A correlation existed between the imazalil concentration necessary to induce such changes in sterol composition and the EC₅₀ values for inhibition of mycelial growth of the different isolates. The reason for the differential effects of imazalil on sterol composition in the variousP. italicum isolates may be due to decreased accumulation of the fungicide in the mycelium and to other yet non-identified mechanisms of resistance.Imazalil remt differentieel de toename in drooggewicht van 10-uur-oude gekiemde sporen van wild-type en DMI-resistente isolaten vanPenicillium italicum in vloeistofcultures van moutextract. De EC₅₀ waarden voor groei van de verschillende isolaten lopen uiteen van 0,005 tot 0,27 g ml^–1. In afwezigheid van het fungicide is in alle isolaten ergosterol het belangrijkste sterol (meer dan 95% van het totaal). DMI-resistentie kan daarom niet in verband staan met deficiëntie van het C-14 demethyleringsenzym in de ergosterol biosynthese. Imazalilbehandeling van mycelium bij concentraties rond de EC₅₀ waarde voor groeiremming, resulteerde bij alle isolaten in een afname van het ergosterolgehalte en een gelijktijdige toename van het gehalte aan 24-methyleen-24,25-dihydrolanosterol. Er bestaat dus een nauwe correlatie tussen de imazalilconcentratie die noodzakelijk is om vergelijkbare veranderingen in sterolsamenstelling te induceren en de EC₅₀ waarde voor remming van myceliumgroei van de verschillende isolaten. De differentiële effecten van imazalil op de sterolsamenstelling van de verschillendeP. italicum isolaten kunnen worden veroorzaakt door verminderde accumulatie van het fungicide in het mycelium en door andere, nog niet geïdentificeerde resistentiemechanismen.     

Optimal cultivar mixtures in focal plant diseases

This paper deals with a study on the effects of mixtures of partially resistant cultivars in focus forming plant diseases. The focus expansion velocity in a mixture of two partially resistant cultivars depends on four parameters, the focus expansion velocity in a crop of cultivar 1 only and in a crop of cultivar 2 only, the net-reproduction in a crop of cultivar 1 and of cultivar 2, respectively. An explicit equation is developed for the damage of a cultivar mixture where the crop is infected by two physiological races of the pathogen. It is shown that cultivar mixtures exist where damage is smaller than in crops of either of the two cultivars only. A simple explicit formula is given on the basis of which it can be decided whether an optimal mixture exists for a given pathosystem.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3-ends, but also the 5-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Postponed germination of Puccinia recondita urediospores deposited on wheat seedlings. II. Infectivity of urediospores after postponed germination

Urediospores ofPuccinia recondita f.sp.tritici were applied to wheat seedlings. Inoculated plants were placed in a growth chamber to expose the spores to dry periods from zero to nine days at near-optimal temperatures (ca 18 °C). The dry period was followed by a wet period varying from 2 to 24 hours for germination of spores and infection of plants. Infection results were subjected to analysis of variance. The main effects dry period, wet period, and temperature were highly significant. The dry period×wet period interaction was significant. The interaction implied that the effects of post-detachment, ripening of germinable spores appeared in their resulting infectivity. There were two forms of post-detachment ripening, a slow ripening during the dry period and a faster ripening during the wet period. The two forms of ripening showed a non-additive compensatory interaction. The effect of post-detachment ripening on infectivity of germinated spores was more pronounced at 15 than, at 18 or 20 °C; the effect was strongest during the first day of the dry period. At dry periods of over 6 days, infectivity of germinated spores decreased, especially at the higher temperatures. Prolonged exposure, of spores to a dry period apparently damages the spores even though they are still able to germinate.Samenvatting Uredosporen van de bruine roest van tarwe (Puccinia recondita f.sp.tritici) werden over het eerste blad van tarwekiemplanten verstoven. De aldus geïnoculeerde planten werden in een klimaatkamer geplaatst bij ca. 18 °C om de sporen bloot te stellen aan droge perioden van 0 tot 9 dagen. De droge perioden werden gevolgd door natte perioden van 2 tot 24 uur om de sporen te laten kiemen en de planten te infecteren. Na de natte periode werden de planten bij verschillende temperature geplaatst om de latente periode en de vorming van sporenhoopjes te bepalen. De hoofdeffecten op de vorming van sporenhoopjes waren zeer significant: duur van de droge periode, duur van de natte periode en temperatuur. Twee vormen van sporenrijping werden gevonden bij rijping vnn sporen, die los op het blad lagen (rijping, buiten de sporenhoopjes), een langzame rijping tijdens de droge periode en een snelle rijping tijdens de natte periode. Deze twee vormen van rijping vertoonden statistische interactie met enige wederzijdse compensatie. Het effect van deze sporenrijping (buiten de sporenhoopjes) op de infectiviteit van gekiemde sporen was bij 15 °C duidelijker dan bij 18 en 20 °C; het effect was het sterkst tijdens de eerste dag van de droge periode. Bij droge periodes langer dan 6 dagen daalde de infectiviteit, vooral bij de hogere temperaturen.     

Effect of maleic hydrazide on parasitic fitness of Puccinia striiformis f. sp. tritici in wheat

As a growth inhibitor of the apical meristem in the plant stem, Maleic hydrazide plays an important role in modulation of plant growth. In this research, Puccinia striiformis f. sp. tritici and its host for reproduction was employed to characterize the effect of maleic hydrazide on parasitic fitness of the strain. Growth inhibition of the secondary leaves of wheat by maleic hydrazide was demonstrated. Results showed that root irrigation by maleic hydrazide at the seedling stage significantly increased the parasitic fitness of Puccinia striiformis f. sp. tritici, leading to the increase in sporulation amount, sporulation period, and germination rate of urediniospores. In addition, the ultrastructure of Puccinia striiformis f. sp. tritici urediniospores was not affected by maleic hydrazide treatment. Out data indicate that the optimal concentration and dose for the use of maleic hydrazide is 0.35 g?l^?1 and 1.5 ml/cm², respectively, facilitating the widespread application in wheat stripe rust studies.     

Effects of fertilization on pests and diseses

The aim of integrated farming systems research is to develop sustainable farming systems from an economic, technical and environmental point of view. Strategies for crop rotation, crop protection and fertilization are briefly discussed in relation to potato production. The effects of different levels of fertilization on the incidence of some pests and diseases of potatoes are discussed in more detail, based on a literature study. Some results of potato production on the three farms for integrated arable farming in the Netherlands are also presented.     

Effect of interrupted leaf wetness periods on suppression of sporulation ofBotrytis allii andB. cinerea by antagonists on dead onion leaves

Saprophytic antagonists were evaluated for suppression of sporulation ofBotrytis allii andB. cinerea on artificially killed segments of onion leaves that were pre-inoculated with the pathogens. During incubation of the antagonisttreated leaf segments in moist chambers, periods of leaf wetness and leaf dryness were alternated to simulate conditions in the field. Interruption of humid conditions with dry periods had a differential effect on antagonists.Alternaria alternata, Chaetomium globosum, Ulocladium atrum andU. chartarum suppressed sporulation ofB. allii almost completely under continuously wet conditions, and when the leaf wetness periods were interrupted with drying periods of 9h imposed 16, 40, and 64 h after the antagonists were applied. When leaf wetness was interrupted 16 h after antagonist application, the number of conidia ofB. allii produced cm^–2 leaf surface after eight days was under the detection limit of 5.2 × 10³ conidia on leaves treated with these antagonists compared to 3.7 × 10⁵ conidia on leaves that were not treated. On the other hand,Gliocladium roseum, G. catenulatum andSesquicillium candelabrum, all highly efficient under continuously wet conditions, were of low to moderate efficiency when leaf wetness periods had been interrupted 16 h after application of the antagonists. The antagonists showed the same differentiation and sensitivity to interrupted wetness periods when tested withB. cinerea.