Assessment of biological and molecular variability between and within field isolates of Plasmodiophora brassicae

Plasmodiophora brassicae is an obligate biotroph that causes clubroot, one of the most damaging diseases of crucifers. Differential cultivars and random amplified polymorphic DNA markers were used to assess the extent of genetic diversity among nine single-gall populations of P. brassicae and 37 single-spore isolates (SSI) derived from four of those field samples. Isolates were classified into eight pathotypes, and each isolate was associated with a unique molecular genotype. Virulence and DNA polymorphisms were detected within and between field isolates, and among SSIs from different pathotypes, hosts and geographical origins. The relatively high level of genetic diversity among field isolates was similar to that among SSIs derived from a single-club field isolate. Molecular and pathogenicity-based classifications were not clearly correlated, but isolates belonging to pathotype P1 were clustered. Two RAPD markers were specific to pathotype P1. The finding that genetic differences can occur in P. brassicae field isolates will be an important consideration in resistance genetic studies and in choosing breeding strategies to develop durable clubroot resistance.     

Suppression of development of Callosobruchus maculatus (F.) (Col.: Bruchidae) in bambara groundnut seeds exposed to solar heat in the Nigerian savanna

The effect of solarization on the development of Callosobruchus maculatus was investigated in the Nigerian savanna during the hot season from April to May, 1999. Development of C. maculatus adult progeny was completely suppressed in seeds of bambara groundnut, Vigna subterranea bearing bruchid eggs or harbouring first and second instar larvae that were exposed to the sun in metal tins, clay pots or polypropylene sacks for 7, 14 or 28 h. Adults of C. maculatus developed only in seeds that were not exposed to sun. Solarization did not have a significant adverse effect on germinability of bambara groundnut seeds.     

Effects of accumulate dead and alive insects in trap on the capture ofTribolium castaneum (herbst)

In the trapping ofTribolium castaneum (Herbst), the activity of traps baited or not baited with aggregation pheromone 4,8-dimethyldecanal is influenced by the presence in the traps of accumulate specimens trapped. This fact has been observed both in trap containing alive or dead specimens. Interactions in the different substances produced by adults of Red Flour Beetle during their communication relationship are involved in this behaviour.The results obtained in the trials suggests that the presence of alive insects in traps is attractive for other free specimens, but under conditions of overcrowding quinone produced by them regulate population density by counteracting the effect of the aggregation pheromone, causing the insects to disperse or suppressing aggregation behaviour.     

Specificity of polycllonal and monoclonal antibodies for the identification of Xanthomonas campestris pv. campestris

Polyclonal and monoclonal antibodies (PCAs and MCAs), produced to whole cells and flagellar extracts ofXanthomonas campestris pv.campestris (Xcc), respectively, were tested for specificity. In immunofluorescence microscopy (IF) the three PCAs tested, reacted at low dilutions with all Xcc strains, some other xanthomonads and non-xanthomonads. At higher dilutions most cross-reactivity with non-xanthomonad strains disappeared. However, the cross-reactivity with strains ofX. c. pv.vesicatoria (Xcv),X. c. pv.amoraciae (Xca) andX. c. pv.phaseoli var. fuscans (Xcpf) remained.Six MCA-producing cell clones viz. 20H6, 2F4, 18G12, 10C5, 17C12 and 16B5 were selected for specificity tests with an enzyme immunoassay (EIA), IF and a dot-blot immunoassay (DBI). None of the MCAs reacted with all Xcc strains in IF and EIA. In DBI, only MCAs 17C12 and 16B5 reacted with all Xcc strains. All six MCAs tested, cross-reacted in one of either tests with other pathovars ofX. campestris, such as Xcv or Xca. The MCAs were also tested in immunoblotting experiments using total bacterial extracts, cell envelope and flagellar extracts. MCAs 20H6, 2F4, 18G12 and 10C5 reacted with the lipopolysaccharide (LPS) of Xcc. MCAs 16B5 and 17C12 reacted with a 39 kilodalton and a 29 kilodalton protein, respectively.It is concluded that the PCAs and MCAs discussed in this study may be used for routine identification and differentiation of (a group of) Xcc strains. The significance of the cross-reactions with other pathovars ofX. campestris needs to be determined by testing seed lots.     

Parry's disease of pears is similar to pear decline and is associated with mycoplasma-like organisms transmitted by Cacopsylla pyricola

Mycoplasma-like organisms (MLOs) were detected by fluorescence microscopy in a number of pear cultivars and pear seedlings growing under a range of different conditions. In the cultivar Conference, MLOs were consistently associated with symptoms of Parry's disease, a decline-like disease of young trees with quince rootstocks. MLO-free pear seedlings rapidly became infected when they were planted outside. Experimental transmission of MLOs to pear seedlings and Conference trees was achieved using pear psyllids, caught outside or raised on infected plants under controlled conditions. Conference trees in an orchard trial remained free from the severe spring symptoms of Parry's disease when they were protected from feeding insects during spring of the previous year. MLOs were graft transmissible, but were not perpetuated by the standard propagation practices of budding or grafting when quince rootstocks were used. Parry's disease appears to be similar to pear decline, an MLO-induced disease well established in several other parts of the world. It is suggested that Parry's disease should be referred to as pear decline.     

Development of a monoclonal antibody-based immunodetection assay for Botrytis cinerea

Three hybridoma cell lines secreting antibodies that specifically recognize Botrytis cinerea and B.fabae , but not B. allii, have been raised from splenocytes of mice immunized with a low molecular-weight fraction (30 kDa) from surface washings of B. cinerea. Antibodies from these cell lines have been used to develop an antigen-based elisa test that will detect B. cinerea in strawberries. This monoclonal antibody immunoassay detection assay should prove useful to both the cut-flower and wine industries. Supernatants from the three specific cell lines recognize mycelial fragments, saline extracts of mycelia and germinating conidia by both ELISA and immunofluorescence. Recognition of non-germinating spores is poor. Supernatants from the specific cell lines did not recognize other fungi normally involved in post-harvest spoilage of fruits and vegetables. Supernatants from KH4 gave the lowest background values with healthy tissue. Indirect evidence from heat, protease and periodate treatment of the antigens indicates that antibodies from all three specific cell lines are recognizing carbohydrate epitopes on a glycoprotein.     

The activity and mobility of triasulfuron in soil as influenced by organic matter,duration, amount and frequency of rain

The influence of different levels of organic matter and of the duration, amount and frequency of rainfall on the biological activity and mobility of triasulfuron were investigated. Biological activity was inversely related to the organic matter content of the soil. Movement of the herbicide down the soil profile, assessed by bioassay, was inversely related to the organic matter content of the soil, and the amount and frequency of rain directly influenced the extent of leaching of the herbicide. Triasulfuron was moved to a depth of at least 22.5 cm when soil columns were exposed to a total of 528 mm of natural rainfall over a period of 6 months. The presence of phytotoxic levels of the herbicide in the surface layers of soil columns 6 months after exposure to outdoor temperatures and natural rainfall suggested that triasulfuron may persist in soils for relatively long periods. Influence de la matiére organique, et de la durée, niveau et fréquence des précipitations sur l'activité et la mobilité du triasulfuron dans le sol L'influence de différentes teneurs en matière organique et de la durée, quantité et fréquence des pluies sur l'activité biologique et la mobilité du triasulfuron a étéétudiée. L'activité biologique est inversement liée avec la teneur enmatière organique du sol. Le mouvement descendant de l'herbicide dans le profil du sol, estimé par bio essai, est inversement lié avec la teneur en matière organique du sol, et la quantité et la fréquence des pluies influencent directement l'importance de la lixiviation de l'herbicide. Le triasulfuron s'est déplacé jusqu'à une profondeur d'au moins 22,5 cm quand les colonnes de sols ont été exposées à un total de 52,8 cm de pluies naturelles sur une période de 6 mois. La présence de taux phytotoxiques d'herbicide dans les couches superficielles des colonnes de sol 6 mois après l'exposition aux températures extérieures et aux précipitations naturelles montre que le triasulfuron peut persister dans 1s sols pendant des périodes relativement longues. Einfluβ der organischen Substanz des Bodens sowie der Niederschlagsdauer, -menge und -frequenz auf Persistenz und Mobilität von Triasulfuron im Boden Die biologische Aktivität sowie die Verlagerung von Triasulfuron im Boden, mit Biotests bestimmt, waren umgekehrt proportional zum Gehalt an organischer Substanz. Die Regenmenge sowie -frequenz beeinflußten die Einwaschung des Herbizids direkt. Bei insgesamt 528 mm natürlichem Niederschlag innerhalb 6 Monaten wurde Triasulfuron in Bodensäulen mindestens 22,5 cm tief eingewaschen. Aus dem Vorhandensein phytotoxischer Mengen des Wirkstoffs in den oberen Schichten der Bodensäulen nach 6 Monaten unter Freilandbedingungen wurde auf relativ hohe Persistenz im Boden geschlossen.     

Phenological patterns in an arable land weed community related to disturbance

The emergence pattern and life cycle of four major species growing in a non–irrigated almond tree grove were analysed in relation to ploughing frequencies and environmental factors. At the community level, the overall emergence pattern was found to be much the same whether or not the soil was disturbed. Nevertheless, soil disturbance in late winter and early spring produced peaks of seedling emergence and brought about an increase in germination. Winter annuals such as Lolium rigidum Gaudin and Diplotaxis erucoides (L.) DC., which emerged in the autumn, started to grow rapidly in winter and spring and were able to pre–empt the environmental resources of the habitat and suppress spring–germinating plants such as Chenopodium album L. and Amaranthus blitoides S. Watson. Late–winter and early–spring disturbances favoured the dominance of summer annuals such as C. album and A. blitoides S. Watson. The different ploughing regimes applied during the first year had effects on plant development and seed production which brought about changes in plant population size during the second year.     

A Japanese powdery mildew isolate with exceptionally large infection efficiency on Mlo-resistant barley

A Japanese field isolate (Race I) of Erysiphe graminis f.sp, hordei was tested on 17 barley lines carrying the mlo powdery mildew resistance gene. Race I produced many successful infections with infection type larger than or equal to 2 on six lines (M66, MC20, SRI, SR7, A tem and Totem), On the remaining 11 lines it reacted with infection type 0. Colony numbers on the Mlo-lines were between 26% and 12 9% of the numbers on a susceptible cultivar Manchuria, These numbers were larger than, or similar to those produced by isolate HL3/5, which has the highest recorded infection efficiency on Mlo-resistant cultivars. The interaction between isolates and lines was highly significant. The isolate GE3, from which HL3/5 was derived by selection, gave rise to occasional colonies corresponding to less than 03° o of the number produced on cv. Manchuria.