Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Fear of predation slows plant-litter decomposition

Aboveground consumers are believed to affect ecosystem functioning by regulating the quantity and quality of plant litter entering the soil. We uncovered a pathway whereby terrestrial predators regulate ecosystem processes via indirect control over soil community function. Grasshopper herbivores stressed by spider predators have a higher body carbon-to-nitrogen ratio than do grasshoppers raised without spiders. This change in elemental content does not slow grasshopper decomposition but perturbs belowground community function, decelerating the subsequent decomposition of plant litter. This legacy effect of predation on soil community function appears to be regulated by the amount of herbivore protein entering the soil.     

Presence of pap-, sfa-, and afa-related sequences in necrotoxigenic Escherichia coli isolates from cattle: evidence for new variants of the AFA family.

Necrotoxigenic Escherichia coli (NTEC) are associated with intestinal and extraintestinal diseases in animals and human beings and produce Cytotoxic Necrotizing Factor 1 (CNF1) or 2 (CNF2). Fourty-three NTEC1, 42 NTEC2, and 32 CNF-negative isolates from cattle were tested by colony DNA hybridization, by plasmid DNA hybridization and by PCR assays for the presence of DNA sequences homologous to the operons coding for fimbrial (PAP/PRS, SFA/FIC, and F17) and afimbrial (AFA/Dr) adhesins of extraintestinal E. coli. Most NTEC1 isolates hybridized with the PAP probes and either the SFA probe (37%) or the AFA probes (49%). Most NTEC2 isolates, in contrast, hybridized with the F17 probe (45%), the AFA probes (19%), or the F17 and AFA probes (22%). A probe-positive plasmid was identified in each of the 19 NTEC2 isolates studied. They all hybridized with the CNF2 toxin probe (Vir plasmids) and most of them with the F17 (6 plasmids) or AFA (7 plasmids) probes. PCR amplification was obtained with 6 of the 11 NTEC isolates tested for the papGII/prsG genes; with all 5 NTEC isolates tested for the sfa and related operons; but with none of the 18 NTEC isolates tested for the afa and related operons. pap-, sfa-, and afa-related sequences are thus present in NTEC isolates from cattle in addition to f17-related operons and may code for adhesins corresponding to specific colonization factors. f17- and afa-related sequences can be located on the Vir plasmids along with the cnf2 gene. Existence of new variants of the AFA/Dr family is evident from the negative results of this family-specific PCR assay.     

Physiological and biochemical consequences of electroimobilisation in conscious sheep

An electroimmobilisation device has been developed to facilitate the automated shearing of sheep, but there is little information on its effects on the body. We have studied its effects on the cardiovascular system and on intermediary metabolism in sheep. Electroimmobilisation caused statistically significant increases in mean arterial pressure, heart rate, cardiac output, renal and hepatic and hindquarter glucose and lactate flux, organ and whole body oxygen flux, hindquarter blood flow and core temperature and decreases in arterial and posterior vena cava blood pH, renal and hepatic blood flow and PaCO2. Notably, no change occurred in PaO2. The metabolic changes demonstrated the capacity of sheep to respond to the increased muscular and cardiovascular work induced by electroimmobilisation. Pulmonary function was not compromised during electroimmobilisation as judged from blood gas changes, and the acid/base changes were rapidly reversed after electroimmobilisation. The recovery to control conditions for all perturbations generally took no longer than 30 min, consistent with a rapid and physiologically adequate reversal by the animal's homeostatic mechanisms.     

Effects of predator hunting mode on grassland ecosystem function

The way predators control their prey populations is determined by the interplay between predator hunting mode and prey antipredator behavior. It is uncertain, however, how the effects of such interplay control ecosystem function. A 3-year experiment in grassland mesocosms revealed that actively hunting spiders reduced plant species diversity and enhanced aboveground net primary production and nitrogen mineralization rate, whereas sit-and-wait ambush spiders had opposite effects. These effects arise from the different responses to the two different predators by their grasshopper prey-the dominant herbivore species that controls plant species composition and accordingly ecosystem functioning. Predator hunting mode is thus a key functional trait that can help to explain variation in the nature of top-down control of ecosystems.     

Imaging of intermittency in ripple-wave turbulence

The dynamics of a fluid surface filled with high-amplitude ripples were studied with a technique (diffusing light photography) that resolves the height at all locations instantaneously. Even when nonlinearities are strong enough to generate a (Kolmogorov) cascade from long wavelength (where energy is input) to shorter wavelength, the resulting turbulent state contains large coherent spatial structures. The appearance of these structures in a thermal equilibrium state (with the same average energy) would be highly improbable.     

A CYLINDER GUIDE FOR USE IN PLATE ASSAY OF PENICILLIN

A guide or template is described which facilitates placing the cylinders on the agar surface in the plate assay of penicillin. The general plan of conducting the assay is discussed and several advantages of the use of the guide are noted. We wish to thank Mr. Albert G. Sterling, instrument maker, for fabricating the guide and the aluminum cylinders.     

Effect of dicyandiamide and hydroquinone on the transformation of urea-nitrogen-15 in soil cropped to wheat

A pot experiment with a loam soil and spring wheat as test crop showed that an application of dicyandiamide (DCD), and especially its combination with hydroquinone (HQ), gave a much larger recovery of soil urea-¹⁵N than treatments based on the application of urea alone or urea plus HQ. Most of the urea-¹⁵N applied to soil was present as organic plus chemically fixed ¹⁵N in the DCD and DCD plus HQ treatments. These two treatments showed the smallest accumulation of urea-derived (NO₃^-+NO₂^-)-¹⁵N. Under well-drained conditions, there was a synergistic effect of the nitrification inhibitor DCD and the urease inhibitor HQ on urea-¹⁵N transformations and the recovery of fertilizer ¹⁵N in soil after the application of urea.     

Reproductive Performance and Reconception of Nellore Cows According to Their Pure‐ or Cross‐Bred Calves

The objective of the present research was to evaluate the effect of a calf's genetic group on the productive and reproductive efficiency of its Nellore dam. Fixed‐time artificial insemination was applied to 800 cows using semen extracted from Nellore, Simmental and Angus Red bulls. Four hundred eleven cows calved, producing 119 Nellore, 103 ½Simmental–½Nellore and 189 ½Nellore–½Angus Red calves. The second mating period, which paired Nellore cows with Simmental bulls, was initiated 10 days after parturitions began and lasted for 5 months. Based on the two successive parturitions, the cumulative parturition rate for calving periods of 3, 4 and 5 months was calculated. Although no significant difference was observed for birth weight among the genetic groups, cross‐bred calves weighed, on average, 10% more than did pure‐bred calves at the age of 205 days. Nellore dams experienced a gestation period that was 7 days longer than did the cross‐bred dams, and the former showed a higher parturition rate at 90 and 120 days of the calving season, but not at 150 days (calving rates of 80.6, 76.4 and 76.2% for mothers of Nellore, ½Nellore–½Angus Red and ½Nellore–½Simmental, respectively, p > 0.05). At 90 and 120 days, Nellore dams produced more kg of calf per mated dam. In conclusion, in a short breeding season, Nellore dams nursing pure‐bred Nellore calves were found to have a higher biological efficiency compared with Nellore dams nursing cross‐bred calves.     

Dynamic oxygen mapping in the root zone by fluorescence dye imaging combined with neutron radiography

Purpose  

The rooted zone of a soil, more precisely the rhizosphere, is a very dynamic system. Some of the key processes are water uptake and root respiration. We have developed a novel method for measuring the real-time distribution of water and oxygen concentration in the rhizosphere as a biogeochemical interface in soil. This enables understanding where and when roots are active in respect to root respiration and water uptake and how the soil responds to it.