Notes on the role of water potential in the pathogenesis of fire blight,caused by Erwinia amylovora

The rate of multiplication of fire blight causing bacteriumErwinia amylovora (Burrill) Winslow et al. depends on the availability of water. Water availability can be quantified by means of the parameter water potential. The relationship between water potential and relative multiplication rate ofE. amylovora was derived from experiments of L. Shaw (1935). This relationship appears to be applicable toE. amylovora in plant tissues and in nectar of flowers.Multiplication and expansion ofE. amylovora in a restricted space, e.g. an intercellular hole, creates a pressure, which may cause schizogenic cavities in soft tissue. Strong tissue, however, may be able to resist this multiplication pressure of the bacteria, so that symptom progression can be prevented. A hypothesis is formulated on how the multiplication pressure may be quantified by means of the parameter water potential. Expansion of bacterial ooze may alo be due to absorbtion of water without increase of dry weight (e.g. a daily cycle of shrinkage and expansion). This expansion may give rise to a swelling pressure, which again may be quantified by means of the parameter water potential.Samenvatting De vermenigvuldigingssnelheid van de bacterie die bacterievuur veroorzaakt (Erwinia amylovora (Burrill) Winslow et al.,), hangt af van de beschikbaarheid van water. De beschikbaarheid van water kan worden gekwantificeerd met de parameter waterpotentiaal. De relatie tussen waterpotentiaal en relatieve vermenigvuldigingssnelheid vanE. amylovora werd afgeleid uit experimenten van L. Shaw (1935, Cornell University Agricultural Experiment Station, Ithaca. Memoir 181). Deze relatie kan zowel worden toegepast op de pathogenese in planteweefsel als op de epifytische ontwikkeling in nektar.Vermenigvuldiging vanE. amylovora in een beperkte ruimte, bijvoorbeeld in een intercellulaire holte, creëert een druk, die tot scheuren van zacht weefsel kan leiden. Sterk weefsel kan de vermenigvuldigingsdruk van de bacteriën vermoedelijk wel weerstaan, zodat uitbreiding wordt verhinderd. In een hypothese wordt beschreven hoe de vermenigvuldigingsdruk zou kunnen worden gekwantificeerd met behulp van de parameter waterpotentiaal. Wateropname door bacterieslijm zonder toename van het drooggewicht (bijvoorbeeld een dagelijkse gang van krimpen en zwellen) kan ook leiden tot een druk. Deze druk kan eveneens worden berekend met de parameter waterpotentiaal.     

‘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.     

Simulation of pressure caused by multiplication and swelling of Erwinia amylovora in intercellular space of host tissue

WhenErwinia amylovora grows, in an intercellular space of a host, and fills this space, further multiplication or swelling may create a pressure, and may cause tearing of host tissue. Theoretically, this bacterial pressure equals the actual water potential of the host tissue minus the water potential at which the bacterial biomass would completely fill the intercellular space, but without exerting pressure.Simulation runs indicate that, when the pressure increases, the extracellular slime ofE. amylovora shrinks by releasing water, thus allowing further production of bacterial dry matter. The slime remains around the bacterial cells as a dense substance, low in water content, having a strong capacity to swell when the pressure induces tearing apart of the host tissue. Simulation runs show that the pressure attains its highest values at evening and night.Some fire blight symptoms that illustrate the evidence of bacterial pressure are discussed.     

Polyploidy in garden tulips. I. A survey of Tulip a varieties for polyploids

Summary As part of a breeding programme for polyploid tulips, chromosome counts have been made on about 600 tulip varieties in the past 10 years. Four tetraploid and 81 triploid varieties were found.An account of the results together with a list of the counts is presented.     

Quantum superposition of macroscopic persistent-current states

Microwave spectroscopy experiments have been performed on two quantum levels of a macroscopic superconducting loop with three Josephson junctions. Level repulsion of the ground state and first excited state is found where two classical persistent-current states with opposite polarity are degenerate, indicating symmetric and antisymmetric quantum superpositions of macroscopic states. The two classical states have persistent currents of 0.5 microampere and correspond to the center-of-mass motion of millions of Cooper pairs.     

Early reactivation of European rivers during the last deglaciation

During the Last Glacial Maximum, the sea-level lowstand combined with the large extent of the Fennoscandian and British ice sheets led to the funneling of European continental runoff, resulting in the largest river system that ever drained the European continent. Here, we show an abrupt and early reactivation of the European hydrological cycle at the onset of the last deglaciation, leading to intense discharge of the Channel River into the Bay of Biscay. This freshwater influx, probably combined with inputs from proglacial or ice-dammed lakes, dramatically affected the hydrology of the region, both on land and in the ocean.     

Identification of candidate genes required for susceptibility to powdery or downy mildew in cucumber

Powdery mildew (PM, caused by Podosphaera fusca) and downy mildew (DM, caused by Pseudoperonospora cubensis) are important diseases of cucumber (Cucumis sativus). Breeding for resistance has been undertaken since the 1940s, but underlying resistance genes have not been functionally analysed yet. The published genome sequence of cucumber catalyses the search for such genes. Genetic studies have indicated that resistances to PM and DM in cucumber are often inherited recessively, which indicates the presence of susceptibility genes (S-genes). Therefore we analyzed the cucumber genome for homologs of functionally proven S-genes known from other plant species. We identified 13 MLO-like genes in cucumber, three of which cluster in Clade V, the clade that contains all known MLO-like susceptibility genes to powdery mildews in other dicots. The expression of one of these three genes, CsaMLO1, located on chromosome 1, was upregulated after PM inoculation. It co-localizes with a QTL for PM resistance previously identified. Also homologs of the susceptibility genes PMR4 and PMR5 are located at this QTL. The second MLO-like gene from Clade V (CsaMLO8) resides in a recessively inherited major QTL for PM resistance at the bottom of chromosome 5, together with a PMR6-like gene. Two major QTL for DM recessive resistance at the top of chromosome 5 co-localize with CsaDMR6-2, which is homologous to the DMR6 susceptibility gene in Arabidopsis. This study has identified several candidate genes for susceptibility to PM and DM in cucumber that may explain QTL for recessively inherited resistance, reported earlier.     

The effect of digital lesions and floor type on locomotion score in Dutch dairy cows

This study describes the effects of floor system, digital dermatitis (DD) and interdigital dermatitis and heel-horn erosion (IDHE) on locomotion performance in 225 dairy cows of 12 commercial dairy herds. Nine herds were kept in cubicle houses with concrete passageways (either solid, slatted, or grooved concrete) and three herds were kept in straw yards. Animals were at most five times examined at monthly intervals for lesion severity of DD and IDHE and for locomotion score. Locomotion score was rated on a scale ranging from 1 to 5 (from normal to severe) and disturbed locomotion (lameness) was defined as a score > or =3. A logistic regression model was used to analyze the 943 observations using lameness (yes/no) as outcome variable. The proportion of observations scored as lame (locomotion score > or =3) increased from 18% 1 month after trimming to 29% at 4 months after trimming. Severe lesions of DD and IDHE were associated with a significantly higher proportion of lame cows. The proportion of animals with disturbed locomotion increased from 16% to 40% as the severity of DD increased and from 17% to 30% with increasing severity of IDHE lesions. Locomotion performance highly differed between the cubicle house and straw yard group. Only 1% of all gaits in straw yard cows were scored as lame, while in cubicle housed cows these percentages varied from 24% to 46% with grooved floors showing the highest average locomotion score. Due to the extreme low incidence of lameness in straw yards, the statistical analysis had to be restricted to observations on concrete floors (n=744). The logistic regression model with lameness (yes/no) as dependent variable and random effects of cow and herd resulted in Odds Ratios for severe DD and IDHE of, respectively, 3.2 and 3.2, both significantly larger than unity. Cows housed at grooved concrete floors showed the highest OR of 6.5 compared to solid concrete floors. Recovery of lameness was poor as disturbance in gait lasted several months.