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.     

正交试验法优化普抗合剂制备工艺

以干浸膏得率和黄芩苷提取率为考核指标,采用正交试验法对普抗合剂水提取醇沉淀制备工艺进行考察.普抗合剂最佳制备工艺方案为加水量10倍,提取2次,每次1 h,55%的乙醇沉淀杂质 .该工艺科学合理,适合于大规模工业生产.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

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.     

The dispersal of Culicoides brevitarsis in eastern New South Wales and associations with the occurrences of arbovirus infections in cattle

Distributions of the vector Culicoides brevitarsis Kieffer (Diptera: Ceratopogonidae) (determined from light trap data) and 2 arboviruses (determined from seroconversions in sentinel cattle) were studied in eastern New South Wales in 1993–94. C brevitarsis was recorded progressively from endemic areas on the north coast, to Nowra on the south coast, and westward to Scone, in the Hunter Valley. C brevitarsis also survived through winter at Paterson, in the Hunter Valley. Its apparently focal reappearance in this marginal area had no obvious effect on the broad pattern of its progression or the dispersal of Akabane and bluetongue viruses. These viruses were first recorded from foci near Coffs Harbour, on the mid-north coast. Their first occurrences at different locations were associated with those of C brevitarsis, but not with each other. The viruses were found only within the recorded limits of the vector's distribution. Delays between the initial occurrence of C brevitarsis and first evidence of virus transmissions at locations ranged from 2 to 7 months. The delays decreased away from the points of focus and were negatively associated with the time of initial occurrence of the vector. Seroconversions to the viruses were related to the presence of C brevitarsis. However, the densities of C brevitarsis had no apparent effect on the initial numbers of cattle seroconverting to either virus. The results support the conclusion that the progressions of C brevitarsis and Akabane and bluetongue viruses were the result of gradual movements by the vector.