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.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Is titan wet or dry?

Titan's dense and cold nitrogen atmosphere contains a small amount of methane under conditions at least approaching those at which one or both constituents would condense. The possibility of methane and nitrogen rain clouds and global methane oceans has been discussed widely. From specific features of radio occultation and other Voyager results, however, it is concluded that nitrogen does not condense on Titan and that Titan has neither global methane oceans nor a global cloud of liquid methane droplets. Certain results indirectly support the conjecture that methane does not condense at any location. However, other considerations favor a methane ice haze high in the troposphere, and liquid and solid methane might exist on the surface and as low clouds at polar latitudes.     

High throughput phenotyping for aphid resistance in large plant collections

ABSTRACT: BACKGROUND: Phloem-feeding insects are among the most devastating pests worldwide. They not only cause damage by feeding from the phloem, thereby depleting the plant from photo-assimilates, but also by vectoring viruses. Until now, the main way to prevent such problems is the frequent use of insecticides. Applying resistant varieties would be a more environmental friendly and sustainable solution. For this, resistant sources need to be identified first. Up to now there were no methods suitable for high throughput phenotyping of plant germplasm to identify sources of resistance towards phloem-feeding insects. RESULTS: In this paper we present a high throughput screening system to identify plants with an increased resistance against aphids. Its versatility is demonstrated using an Arabidopsis thaliana activation tag mutant line collection. This system consists of the green peach aphid Myzus persicae (Sulzer) and the circulative virus Turnip yellows virus (TuYV). In an initial screening, with one plant representing one mutant line, 13 virus-free mutant lines were identified by ELISA. Using seeds produced from these lines, the putative candidates were re-evaluated and characterized, resulting in nine lines with increased resistance towards the aphid. CONCLUSIONS: This M. persicae-TuYV screening system is an efficient, reliable and quick procedure to identify among thousands of mutated lines those resistant to aphids. In our study, nine mutant lines with increased resistance against the aphid were selected among 5160 mutant lines in just 5 months by one person. The system can be extended to other phloem-feeding insects and circulative viruses to identify insect resistant sources from several collections, including for example genebanks and artificially prepared mutant collections.     

A serological survey of cattle in the Thames-Coromandel district of New Zealand for antibodies to Ross River virus

AIM: To determine if cattle exposed to the southern saltmarsh mosquito (SSM), Aedes camptorhynchus, in the Thames-Coromandel district of New Zealand had been exposed to Ross River virus (RRV).

METHODS: A purposive sampling design was used to test cattle from seven farms located in close proximity to four sites infested with A. camptorhynchus in the Thames-Coromandel district. Sera from 207 cattle were tested for antibodies to RRV, using an ELISA and confi rmatory virus neutralisation test (VNT) as the gold standard.

RESULTS: All 207 cattle tested negative for antibodies to RRV using the ELISA and VNT.

CONCLUSIONS: This study found no evidence of exposure to RRV in cattle in locations in the Thames-Coromandel district of New Zealand where populations of SSM were present.     

Obituary: James Thomas Kelly,BVSc

Extract

On 28 August 1987 James Thomas Kelly, veterinary surgeon, of Milson Line, Palmerston North, died at 68 years of age.     

Early Pregnancy Diagnosis by Serum Progesterone and Ultrasound in Sheep Carrying Somatic Cell Nuclear Transfer-Derived Pregnancies

Early pregnancy diagnosis and monitoring play an important role following embryo transfer in sheep. The aims of the current study were to investigate (i) the pattern of serum progesterone profiles in sheep carrying somatic cell nuclear transfer (SCNT)‐derived (clone) pregnancies, and (ii) the frequency of pregnancy loss during development following SCNT embryo transfer. Sheep SCNT embryos were made using standard nuclear transfer techniques. Day 7 embryos were surgically transferred to oestrus‐synchronized recipients (n = 27). As a control, normal fertile ewes (n = 12) were bred by natural breeding. Serum was collected from all the ewes on the day of estrus (day 0 sample), 7 days post‐estrus (day 7 sample) and 19 days post‐estrus (day 19 sample) and every 10 days thereafter until lambing or pregnancy loss occurred. Serum progesterone (P4) was assessed using enzyme immunoassay. Pregnancy was confirmed by ultrasound scanning on day 35 of pregnancy followed by subsequent scanning every 10 days. In control ewes, pregnancy rate on day 35 was 83.3% (10/12), whereas in the ewes that received SCNT embryos, it was 22.2% (6/27; p < 0.05). The day 45 pregnancy rate in the control ewes was 83.3%, whereas in the SCNT embryo recipients it was 11.0% (p < 0.05). Hormone analysis revealed that SCNT embryo recipients exhibited a significantly lower P4 profiles at different time points in pregnancy compared to controls (p < 0.05). This study highlights the use of serum progesterone in combination with ultrasound for the investigation of embryo loss and crucial times during development of normal and SCNT embryos in sheep. Further, the serum P4 levels directly reflect the degree of placental development in these two groups.     

Voyager radio science observations of neptune and triton

The Voyager 2 encounter with the Neptune system included radio science investigations of the masses and densities of Neptune and Triton, the low-order gravitational harmonics of Neptune, the vertical structures of the atmospheres and ionospheres of Neptune and Triton, the composition of the atmosphere of Neptune, and characteristics of ring material. Demanding experimental requirements were met successfully, and study of the large store of collected data has begun. The initial search of the data revealed no detectable effects of ring material with optical depth tau [unknown] 0.01. Preliminary representative results include the following: 1.0243 x 10(26) and 2.141 x 10(22) kilograms for the masses of Neptune and Triton; 1640 and 2054 kilograms per cubic meter for their respective densities; 1355 +/- 7 kilometers, provisionally, for the radius of Triton; and J(2) = 3411 +/- 10(x 10(-6)) and J(4) = -26(+12)(-20)(x10(-6)) for Neptune's gravity field (J>(2) and J(4) are harmonic coefficients of the gravity field). The equatorial and polar radii of Neptune are 24,764 +/- 20 and 24,340 +/- 30 kllometers, respectively, at the 10(5)-pascal (1 bar) pressure level. Neptune's atmosphere was probed to a pressure level of about 5 x 10(5) pascals, and effects of a methane cloud region and probable ammonia absorption below the cloud are evident in the data. Results for the mixing ratios of helium and ammonia are still being investigated; the methane abundance below the clouds is at least 1 percent by volume. Derived temperature-pressure profiles to 1.2 x 10(5) pascals and 78 kelvins (K) show a lapse rate corresponding to "frozen" equilibrium of the para- and ortho-hydrogen states. Neptune's ionosphere exhibits an extended topside at a temperature of 950 +/- 160 K if H(+) is the dominant ion, and narrow ionization layers of the type previously seen at the other three giant planets. Triton has a dense ionosphere with a peak electron concentration of 46 x 10(9) per cubic meter at an altitude of 340 kilometers measured during occultation egress. Its topside plasma temperature is about 80 +/- 16 K if N(2)(+) is the principal ion. The tenuous neutral atmosphere of Triton produced distinct signatures in the occultation data; however, the accuracy of the measurements is limited by uncertainties in the frequency of the spacecraft reference oscillator. Preliminary values for the surface pressure of 1.6 +/- 0.3 pascals and an equivalent isothermal temperature of 48 +/- 5 K are suggested, on the assumption that molecular nitrogen dominates the atmosphere. The radio data may be showing the effects of a thermal inversion near the surface; this and other evidence imply that the Triton atmosphere is controlled by vapor-pressure equilibrium with surface ices, at a temperature of 38 K and a methane mixing ratio of about 10(-4).