In vitro selection of sterol-biosynthesis-inhibitor (SBI)-resistant mutants in Monilinia fructicola (Wint.) Honey

The inherent resistance risk forMonilinia fructicola against sterol-biosynthesis inhibitors (SBIs) was estimated inin vitro andin vivo laboratory studies. Several mutant strains were selected on media amended with the triazole fungicides penconazole, etaconazole or the morpholine fungicide fenpropimorph.The potential forM. fructicola to develop resistance to the triazoles or to the morpholines was similar.The level of resistance attained did not differ for the two classes of fungicides after a single cycle of treatment with nitrosoguanidine (NTG). Attemps to select mutants with a higher level of resistance to penconazole after successive mutagenic treatments were successful. Most of the mutants were less fit than wild-type strains. Mutants with a low level of resistance had an almost normal mycelial growth rate, whereas growth of mutants with a higher level of resistance was significantly reduced. Spore production was highest in the wild-type strains, similar to the latter in a few resistant strains and less in most others. Only one mutant with an intermediate level of resistance could successfully compete in a mixed population with a wild type strain during successive infection cycles on peaches. Resistance was not stable in highly resistant mutants. Cross resistance to the inhibitors of 14-methylsterol demethylation (DMIs) tested was confirmedin vitro andin vivo for all mutant strains. One DMI-resistant mutant was also resistant to fenpropimorph and two fenpropimorph-resistant mutants were resistant to penconazole.     

Colonization of rose by Sphaerotheca pannosa (Wallr.) Lév. var. rosae Wor.

Colonization of rose by powdery mildew (Sphaerotheca pannosa) is described in terms of mycelium growth, conidiophore production and sporulation in time. The data used are gathered during different years, put together and treated by means of graphic models. Colonies could be separated into fast and slow growing colonies. Colonies initiated on leaves of increasing age showed a decreasing growth rate. Production of conidiophores and conidia started on the same day, and the relative activity of conidiophore production reached its maximum 6 days after the end of the latency period, followed 1 day later by the maximum activity of conidium production. Both conidiophore and conidium production continued for a long time at a low level. The effect of leaf age on conidiophore production found expression in differences in production rate during the first days of colony development and in final production levels. Observations on naturally infected leaves in an outdoor experiment showed a rapid decrease of sporulation on leaves of 10 days and older. Highest percentages of sporulating leaf area were observed on leaves between 7 and 10 days old.     

Inactivation of soil-borne plant pathogens during small-scale composting of crop residues

Samples of heavily infested crop residues were incorporated in static compost heaps (2.5–4.6 m³) of the Indore type. Temperature increased to 50–70°C within 6 days depending on the type of crop residues used and the location within the heap. The heat phase (>40 °C) lasted 2–3 weeks and was followed by a c. 5-months maturation phase (<40 °C). Among the 17 pathogens tested, onlyOlpidium brassicae and one of the four formae speciales ofFusarium oxysporum that were tested survived composting, but also their inoculum was greatly reduced.Survival during specific phases of composting was studied by incorporation and retrieval of samples at various stages of the process.F. oxysporum f. sp.melonis was completely inactivated andO. brassicae andPlasmodiophora brassicae were almost completely inactivated during the short heat phase. The three pathogens survived the long-lasting maturation phase without loss of viability. Heat evolved during composting was found to be the most important factor involved with sanitation of crop residues. The possible involvement of fungitoxic conversion products and microbial antagonism is discussed.     

A formation mechanism for catalytically grown helix-shaped graphite nanotubes

The concept of a spatial-velocity hodograph is introduced to describe quantitatively the extrusion of a carbon tubule from a catalytic particle. The conditions under which a continuous tubular surface can be generated are discussed in terms of this hodograph, the shape of which determines the geometry of the initial nanotube. The model is consistent with all observed tubular shapes and explains why the formation process induces stresses that may lead to "spontaneous" plastic deformation of the tubule. This result is due to the violation of the continuity condition, that is, to the mismatch between the extrusion velocity by the catalytic particle, required to generate a continuous tubular surface, and the rate of carbon deposition.     

The role of autotoxins from root residues of the previous crop in the replant disease of asparagus

Replant disease is a common phenomenon in asparagus growing in the Netherlands. It is distinct from the decline phenomenon reported from many other asparagus producing area’s. The involvement of autotoxins from root residues of former asparagus crops was evaluated. Residues of aspar agus roots decompose extremely slowly. At two locations, each with fields where asparagus production was terminated 1 and 10 years before, biomass of root residues was 4180 and 11060 kg dw ha^?1 after 1 year and 420 and 1140 kg dw ha^?1 after 10 years.Although 10-year-old residues were for the greater part decomposed, crude aqueous extracts inhibited root growth of asparagus seedlings significantly and even more of garden cress. In root observation boxes with field soil mixed with non-sterilized or sterilized asparagus root fragments, growth of secondary roots was inhibited. Non-sterilized strawberry roots did not inhibit root growth, suggesting that effects of organic matter were not involved. In a pot experiment, sterilized asparagus root fragments inhibited root growth when added at a rate of 20 g1^?1, but not a 2 g1^?1 Addition of non-sterilized root fragments strongly inhibited root growth at both levels. This was probably due to simultaneous infection byFusarium oxysporum present in these residues. When an asparagus field is replanted, the amount of root residues left behind in soil after termination of the crop in the previous season is about 2 g dw 1^?1, that corresponds to approx. 11000 kg dw ha^?1. This level is too low for considering direct growth inhibition by autotoxins as a major factor. Their possible indirect effects are briefly discussed.     

小麦对条锈病的水平抗病性研究初报

将小麦品种采用随机区组的田间设计种于小种病圃中,以鉴定和测定其水平抗性。根据各个“品种——小种”组合的相对病情指数的方差分析的结果,有些品种,如农大311、西农6028和丰产3号,其品种——小种互作高度显著,被鉴定为具有垂直抗性,另外一些品种,对试验所用的小种并无专化性,并且在大田生产中已显出20多年的持久的中度抗性,初步推测为属水平抗性。在后一类中,有些品种,如平原50,表现为中等反应型,其余则属于呈典型感病的迟锈慢锈类型。在迟锈慢锈品种的病指的对数矫正值(logits)和病圃的锈病强度(以感病对照品种的病指表之)的对数矫正值之间发现了直线回归。     

Decay of female sexual behavior under parthenogenesis

A laboratory strain of Drosophila mercatorum has existed for 20 years without males and therefore without natural selection operating to maintain the genetic basis of female mating behavior. The females of this strain have recently experienced a genetic impairment of mating capacity. This observation exemplifies the mode of evolution of vestigial characters and supports Muller's theory that random mutation will tend to destroy the genetic basis of a character from which selection has been removed.     

‘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.