Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy.

The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.     

Biological evidence for a 1:1 Ca2+:glyphosate association in deposit residuals on the leaf surface of barley

It has long been known that calcium ion antagonizes glyphosate, but it was not clear whether the stoichiometry of their interaction is 1:1 or 1:2. Two independent methods were used to determine which stoichiometry was the most probable. First, dose–response curves of barley ( Hordeum vulgare L.) plants treated with glyphosate were determined in the presence of 0, 1.25, 2.5, 5 and 10 mM CaCl₂. The doses of 'free' glyphosate (=not inactivated by calcium ion) were computed using the assumptions of 1:1 and 1:2 stoechiometries. The response curves were redrawn as a function of 'free' glyphosate. Analysis showed that the 1:2 hypothesis could be rejected, whereas the 1:1 hypothesis was highly probable. Second, kinetics of glyphosate penetration into barley leaves were determined in the presence of 0, 2.5 and 10 mM CaCl₂. The concentrations of 'free' glyphosate were computed as above. The kinetics of glyphosate penetration at these concentrations were established and were compared to the kinetics of glyphosate penetration in the presence of CaCl₂. Again, the 1:2 hypothesis was rejected, whereas the 1:1 hypothesis was more probable. These results strongly suggest that the stoichiometry of the Ca²⁺:glyphosate association is 1:1 in deposit residuals on the leaf surface.     

Cutting weeds with a CO2 laser

Stems of Chenopodium album . and Sinapis arvensis . and leaves of Lolium perenne . were cut with a CO₂ laser or with a pair of scissors. Treatments were carried out on greenhouse-grown pot plants at three different growth stages and at two heights. Plant dry matter was measured 2 to 5 weeks after treatment. The relationship between dry weight and laser energy was analysed using a non-linear dose–response regression model. The regression parameters differed significantly between the weed species. At all growth stages and heights S. arvensis was more difficult to cut with a CO₂ laser than C. album . When stems were cut below the meristems, 0.9 and 2.3 J mm⁻¹ of CO₂ laser energy dose was sufficient to reduce by 90% the biomass of C. album and S. arvensis respectively. Regrowth appeared when dicotyledonous plant stems were cut above meristems, indicating that it is important to cut close to the soil surface to obtain a significant effect. When cutting L. perenne plants with 2-true leaves at a height of 2 cm from the soil surface with a laser, the biomass decreased significantly compared with plants cut by scissors, indicating a delay in regrowth. This delay was not observed for the dicotyledonous plants nor for the other growth stages of L. perenne .     

Metabolism, uptake and translocation of 14C-paraquat in resistant and susceptible biotypes of Crassocephalum crepidioides (Benth.) S. Moore

The metabolism, uptake and translocation of paraquat in resistant (R) and susceptible (S) biotypes of Crassocephalum crepidioides (Benth.) S. Moore (redflower ragleaf) at the 10-leaf stage was investigated. A study on the properties of leaf surface was carried out to examine the relationship between leaf surface characters and paraquat absorption. The extractable paraquat was not metabolized by the leaf tissue of either the resistant or susceptible biotypes. Differential metabolism, therefore, does not appear to play a role in the mechanism of resistance. Both biotypes did not show any significant difference in the amount of cuticle and trichome densities. Furthermore, both biotypes are identical in the structure of stomata, trichomes and epicuticular wax. The results of the leaf surface studies are in agreement with the findings of the uptake study. Both biotypes demonstrated no significant difference in absorption between the resistant and susceptible biotypes. However, 10% of the absorbed ¹⁴C-paraquat into the S biotype was translocated basipetally, but not in the R biotype. The results of this study suggest that in C. crepidioides , differential translocation may contribute to the mechanism of resistance at the 10-leaf stage.     

Ribosomal DNA Sequence Comparisons of Colletotrichum Graminicola from Turfgrasses and other Hosts

The 5.8S ribosomal gene and the flanking internal transcribed spacers (ITS) 1 and 2 from Colletotrichum graminicola isolates causing anthracnose disease of Agrostis palustris and Poa species were sequenced. Although bootstrap support was not high, two major groups were observed with both UPGMA and parsimony algorithms, one containing isolates from A. palustris and another with isolates from Poa spp. The ITS sequences were also compared with those of isolates of C. graminicola and C. sublineolum from Sorghum spp., Zea mays and Rottboellia cochinchinesis as well as other Colletotrichum species. Except for one isolate from P. annua in Texas, the ITS1 and ITS2 sequences of turfgrass isolates always grouped separately from C. graminicola or C. sublineolum from non-turfgrass hosts with high bootstrap support. ITS sequences of the turfgrass isolates were more similar to those of other species of Colletotrichum, such as C. coccodes and C. dematium, than they were to C. graminicola isolates from other hosts. Turfgrass isolates have ITS sequences which are not identical to those of isolates from Zea mays and Sorghum species demonstrating diversity among fungi conventionally classified as C. graminicola.     

Optimization of production of nucleopolyhedrovirus ofHelicoverpa armigera throughout larval stages

Studies were carried out to optimize production of nucleopolyhedrovirus of American bollworm,Helicoverpa armigera (Hübner), by treating larvae individually with an inoculum dose that allowed maximal larval growth and also gave the highest occlusion bodies (OB) yield/larva. The maximum virus yield of 12.2x 10⁹ OB/larva was obtained when 6-day-old larvae were fed individually with a dose of 1 x 10³ OB. Topical spiracular treatment of larvae as old as 8 days with 10 μ of 2x 10⁷ OB ml^-1 gave the highest yield, of 15.2x 10⁹ OB from 13-day-old larvae, of 12.8x 10⁸ OB from prepupae and of 1.49x 10⁸ OB from pupae at the time of their death. These studies showed that dietary inoculum is the best route for 6-day-old larvae and topical spiracular treatment is the best for 8-day-old larvae.     

Biological control of botrytis leaf blight of onion byGliocladium roseum applied as sprays and with fabric applicators

Gliocladium roseum (5×10⁶ conidia ml^–1) and chlorothalonil (Bravo 500) were compared in two field tests for effectiveness in suppressing leaf blight caused byBotrytis squamosa in cooking onions. The biological control agent and fungicide were applied with sprayers and with specially designed fabric applicators that were mounted interchangeably on an aluminium carriage with bicycle wheels. In the applicators, inoculum and fungicide gravitated from a reservoir down curtains of denim strips and onto portions of onion leaves contacted by the strips. Initial applications were timed using a leaf blight forecasting system (BOTCAST) and three or four subsequent applications were made at weekly intervals. When compared to water checks,G. roseum applied as sprays or by applicators reduced density of leaf spots by about 50–58% during middle and late stages of epidemics. The antagonist was about half as effective as chlorothalonil in suppressing density of spots, regardless of application method. The applicators delivered inoculum in similar densities to proximal and distal halves of leaves, and used at least 35% less volume of inoculum compared to the sprayers. Density of inoculum on leaves treated by applicators was about the same as in spray-treated leaves when canopies were sparse, but less when canopies were dense. It is concluded thatG. roseum has good potential for controlling leaf blight sufficiently to avoid economic yield losses.     

The role ofPseudomonas spp. and competition for carbon,nitrogen and iron in the enhancement of appressorium formation byColletotrichum coccodes on velvetleaf

Colletotrichum coccodes is currently being investigated as a mycoherbicide against the weed velvetleaf (Abutilon theophrasti). Two isolates ofPseudomonas spp. (Ps2 and Ps5) reduced the percentage of germ tubes and increased appressorial formation ofC. coccodes on detached leaves of velvetleaf. A study was conducted to see whether this effect could be attributed to competition for nutrients or iron betweenC. coccodes andPseudomonas spp. Ps2 and Ps5 had no effect on early spore germination, but reduced the percentage of germ tubes at 24 and 30 h, compared to the nontreated control. This reduction was diminished by the addition of nutrients but not Fe³⁺. Ps2 and Ps5 stimulated the formation of dark-coloured appressoria without germ tubes (AWGT), but this stimulation was diminished by the addition of nutrients or Fe³⁺. Germ tube branching at 30 h was also inhibited by the bacteria, but was not diminished by the addition of nutrients or iron. EDTA stimulated conidial germination at 10 h, which was reduced by the addition of Fe³⁺. However, EDTA did not stimulate the formation of appressoria (AWGT). These results suggest that the reduction in the percentage of germ tubes and the increase in the percentage of appressoria induced by the bacteria may be due to the competition for carbon or nitrogen. Iron competition may also be involved in the stimulation of appressorial formation, but not in the reduction in germ tube percentage and branching. Phylloplane bacteria may compete for carbon, nitrogen and iron, limiting the saprophytic phase of the pathogen on the phylloplane and accelerating the development of the parasitic phase. This may enhance the field efficacy ofC. coccodes as a biocontrol agent against velvetleaf.     

Pathogenic specialization in Melampsora epitea var. epitea on Salix

Eighteen isolates of the rust fungus Melampsora from different locations in England were tested for pathogenicity to a large range of willow clones ( Salix spp.) in experiments involving inoculation of leaf discs. Seventeen of the isolates were of leaf-infecting M. epitea var. epitea , 16 of which represented forms which alternated on Larix and one of which alternated on Ribes . The remaining isolate was of uncertain identity. Two experiments were carried out. In the first experiment, 24 willow clones consisting of 20 Salix species and interspecific hybrids were inoculated with eight isolates from clones of S. viminalis . In the other experiment, 77 clones from 57 species or hybrids were inoculated with 10 isolates from several Salix spp. The M. epitea var. epitea isolates from S. viminalis clones were all similarly pathogenic, whilst most of the other isolates expressed distinct host specificity. Eight distinct pathotypes were recognized within M. epitea var. epitea . All except one of these alternated on Larix and could be assigned to three formae speciales which had been reported previously in Europe: four pathotypes to f.sp. larici-epitea typica , two to f.sp. larici-retusae , and one to f.sp. larici-daphnoides . The Ribes -alternating pathotype of M. epitea var. epitea infected only S. purpurea . Nine differential willow hosts are proposed as reference clones to distinguish between the larch-alternating pathotypes, the Ribes -alternating rust and the 'stem-infecting' form. The extent of pathogenic variation encountered amongst sexually reproducing rusts suggests that more pathotypes probably exist and will arise in future in response to selection given by long-term clonal willow plantings.