Molecular, physiological and pathological characterization of Corynespora leaf spot fungi from rubber plantations in Sri Lanka

The plant pathogenic fungus Corynespora cassiicola causes a severe leaf spot disease on more than 70 host plant species including Hevea brasiliensis . Genetic variability in 32 isolates of C. cassiicola collected from diverse hosts and locations in Sri Lanka and Australia was assessed using restriction fragment length polymorphism (RFLP) analysis of the internal transcribed spacer (ITS) region of ribosomal DNA and random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis of total fungal DNA. Amplified ITS fragments from all 32 C. cassiicola isolates exhibited an identical size, and restriction analysis with seven different restriction endonucleases revealed identity in all of the detected DNA fragments. This finding of high genetic relatedness was further supported by the cloning and DNA sequencing of the ITS2 region from one Sri Lankan and one Australian isolate. However, RAPD-PCR profiles generated by 15 oligonucleotide decamer primers revealed significant polymorphism between groups of organisms. Genetic relationships among the isolates were determined by cluster analysis of the RAPD-PCR data and seven different RAPD groups were identified. Isolates showed strong correlations between the assigned RAPD group and the location and host plant genotype from which the isolate was collected. Correlations were also observed between the RAPD group, growth of the isolate and pathogenicity on different plant hosts.     

Pregnancy rates of mares inseminated with semen cooled for 18 hours and then frozen

The ability to ship cooled stallion sperm for subsequent freezing at a facility specializing in cryopreservation would be beneficial to the equine industry. Stallion sperm has been centrifuged, cooled to 5 degrees C for 12 h, and frozen without a detrimental effect on motility in a previous study; however, no fertility data were available. Experiment 1 compared the post-thaw motility of sperm cooled for 18 h at 15 or 5 degrees C at either 400 or 200 x 10(6) sperm/mL and then frozen. Storage temperature, sperm concentration, or the interaction of temperature and concentration had no effect on total (TM) and progressive motility (PM) after cooling. Post-thaw TM and PM were higher for control than (P < 0.05) for treated samples. There was no difference in post-thaw TM and PM due to temperature or concentration. Experiment 2 further evaluated procedures for cooling before freezing. Ejaculates were either cooled to 5 degrees C for 18 h and centrifuged, centrifuged at room temperature and then cooled to 5 degrees C for 18 h before freezing, or centrifuged and frozen immediately (control). There was no difference among treatments on post-thaw TM or PM. In Exp. 3, mares were inseminated with semen that had been extended in skim milk-egg yolk without glycerol, centrifuged, resuspended at 200 x 10(6) sperm/mL, cooled to 5 degrees C for 18 h, and then frozen or not cooled for 18 h before freezing (control). Pregnancy rates did not differ for mares receiving semen cooled and then frozen (21 of 30, 70%) or semen frozen directly without prior cooling (16 of 30, 53%). In summary, a procedure was developed for cooling stallion sperm for 18 h before freezing without a resultant decrease in fertility.     

Influence of temperature,humidity duration and growth stage on the infection and mycotoxin production by Fusarium langsethiae and Fusarium poae in oats

High occurrence of Fusarium poae (FP) and Fusarium langsethiae (FL) and their mycotoxins nivalenol (NIV) and T-2/HT-2 have been observed in Swiss oats. Early prediction of mycotoxin levels is important for farmers and the cereal industry to minimize the risk of contaminated food and feed. Therefore, climate chamber experiments were conducted to investigate the influence of different temperatures (10, 15, 20 °C) and durations (4, 8, 12 h) at 99% relative humidity (RH) on the infection of oats with FP and FL. In addition, to discover the most susceptible period of oats, artificial FL inoculations were conducted at different growth stages. Field experiments were performed to observe the dispersal of these fungal species within the field and to investigate the weather conditions that influence the dispersal. The climate chamber experiments revealed higher contamination with NIV and T-2/HT-2 in the 10 °C treatments and with a prolonged humidity duration of 12 h 99% RH. Inoculations of oat plants at early (DC 61) and mid (DC 65) anthesis, led to higher FL infection and T-2/HT-2 accumulation in the grains compared with treatments at earlier growth stages, which might be due to an increased susceptibility during anthesis. No indication for spore dispersal was observed in the field experiments. The results obtained, together with the cropping factors that influence infection and mycotoxin production, could be used as a first step in developing forecasting models to predict the contamination of oats with the mycotoxins NIV and T-2/HT-2.     

Abscisin II, an Abscission-Accelerating Substance from Young Cotton Fruit

Crystalline abscisin II, with a tentative molecular formula of C(15)H(20)O(4), has been isolated from young cotton fruit. It accelerates abscission when applied in amounts as low as 0.01 microg per abscission zone. It inhibits indoleacetic acid-induced straight growth of Avena coleoptiles but has no gibberellin activity on dwarf maize.     

Metabolism of 15N-14C-acetylurea in the sheep]

3 male sheep (phi 48.3 kg) were fed a semisynthetic diet containing acetyl urea as sole protein source and 15N-14C labelled acetyl urea (urea-C labelled) by intraruminal tube. A half life period of 4 hrs was established for the removal of labelled acetyl urea from the TCE-soluble portion of the ruminal fluid. The degree of 14C labelling in ruminal proteins was very low whereas the extent of 15N labelled protein synthesis was quite marked reaching a maximum between the 18th and 24th hour of experiment. The steepest rise of 15N incorporation into ruminal proteins was found to occur between 8 to 12 hrs after start of the experiment, i.e. at the time of peak level of 15N returned from 15N urea via the rumino-hepatic circulation. 23.3% of the amount of 14C activity administered (mean of all 3 experimental animals) was excreted through respiration. The curve patterns of both isotopes in the TCE soluble portion of the ruminal fluid were similar to that of the degasified TCE soluble portion of the blood blasma. At the peak time (8 hrs) a concentration of the nitrogen isotope of about 4 atom% excess of 15N was observed. The level of 14C labeling in blood plasma proteins was insignificant when compared with that of 15N labelling. The ratio at the peak time was 1:10; the same ratio was found for ruminal proteins. From this it can be concluded that the process of labelling of blood plasma proteins proceeds mainly through microbial protein synthesis. Sheep I and III excreted an average of 60.6% of 14C activity and 57.0% of the administered excess of 15N in the urine. 6 hrs after the beginning of the experiment 81% of the amount of urinary 14C activity was found to occur as acetyl urea; after 48 hrs this amount had decreased to 50%. All experimental sheep excreted a urinary sediment consisting mainly of acetyl urea. The level of faecal 14C excretion (1.4%-2.9% of the amount administered) was considerably lower than that of 15N excretion (9.1%--15.6% of the administered dose). The TCE soluble fraction of the faeces contained up to 2% of the 14C dose and 3% of the 15N dose. The true digestibility data of 15N from 15N acetyl urea varied between 96.4% and 98.2%. An average of 40.9% was obtained for the 15N balance over the 7-day trial period.