Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.     

Genetic control of acquired high temperature tolerance in winter wheat

Summary The development of high temperature-tolerant wheat (Triticum aestivum L.) germplasm is necessary to improve plant productivity under high-temperature stress environments. The quantification of high temperature tolerance and the characterization of its genetic control are necessary for germplasm enhancement efforts. This study was conducted to determine the genetic control of acquired high temperature tolerance in common bread wheat cultivars. Reduction of 2,3,5-triphenyltetrazolium chloride (TTC) by heat-stressed seedling leaves was used as a quantitative measure to characterize acquired high temperature tolerance. Eleven-day-old seedlings of 20 F₁ progeny produced through a complete 5×5 (Payne, Siouxland, Sturdy, TAM W-101, and TAM 108) diallel mating design were acclimated at 37° C for 24 hours, followed by a 2-hour incubation at 50° C. Under these test conditions, acquired high temperature tolerance ranged from a high of 75.7% for the genotype TAM W-101 × TAM 108, to a low of 37.3% for the genotype Payne × Siouxland. Partitioning of genotypic variance revealed that only the general combining ability component effect was statistically highly significant, accounting for 67% of the total genotypic variation. These results suggest that enhancing the level of high temperature tolerance in wheat germplasm is feasible utilizing existing levels of genetic variability and exploiting additive genetic effects associated with high temperature tolerance.Contribution of the Texas Tech College of Agric. Sci. Journal no T-4-386. This work was supported by USDA specific agreement No. 58-7MNI-6-114 from the Plant Stress and Water Conservation Laboratory, USDA-ARS, Lubbock, Texas, USA     

Effects on nutrients and on grain quality in spring wheat crops grown under elevated CO2 concentrations and stress conditions in the European, multiple-site experiment ‘ESPACE-wheat’

Nutrient element concentrations and grain quality were assessed in spring wheat grown under elevated CO₂ concentrations and contrasting levels of tropospheric ozone at different nitrogen supply rates at several European sites. Carbon dioxide enrichment proved to affect nutrient concentrations in a complex manner. In green leaves, all elements (with exception of phosphorus and iron) decreased. In contrast, effects on the element composition of grains were restricted to reductions in nitrogen, calcium, sulphur and iron. Ozone exposure resulted in no significant effects on nutrient element concentrations in different tissues in the overall analysis. The nitrogen demand of green tissues was reduced due to CO₂ enrichment as shown by reductions in the critical leaf nitrogen concentration and also enhanced nitrogen use efficiency. Reductions in the content of ribulose-bisphosphate carboxylase/oxygenase and repression of the photorespiratory pathway and reduced nitrogen allocation to enzymes driving the photosynthetic carbon oxidation cycle were chiefly responsible for this effect. Thus, nitrogen acquisition by the crop did not match carbon acquisition under CO₂ enrichment. Since crop nitrogen uptake from the soil was already completed at anthesis, nitrogen allocated to the grain after anthesis originated from vegetative pools—causing grain nitrogen concentrations to decrease under CO₂ enrichment (on average by 15% when CO₂ concentrations increased from 360 to 680 μmol mol⁻¹). Correspondingly, grain quality was reduced by CO₂ enrichment. The Zeleny value, Hagberg value and dry/wet gluten content decreased significantly with increasing [CO₂]. Despite the beneficial impact of CO₂ enrichment on growth and yield of C₃ cereal crops, declines in flour quality due to reduced nitrogen content are likely in a future, [CO₂]-rich world.     

Ectomycorrhizal fungi identification in single and pooled root samples: terminal restriction fragment length polymorphism (TRFLP) and morphotyping compared

We describe here the results of a study conducted to evaluate a terminal restriction fragment length polymorphism (TRFLP) approach targeting rRNA genes for determination of ectomycorrhizal (ECM) communities colonizing the roots of loblolly pine (Pinus taeda L.). Root tips separated from soil cores were classified according to morphological characteristics and DNA was then extracted from each group of morphotyped tips. Labeled primers were used to generate terminal restriction fragments (TRF) for molecular fingerprinting of root colonizing fungi and to determine how well TRFLP could be used to discriminate between ectomycorrhizal types. Morphotypes generally correlated well with specific TRFs and sequence analysis confirmed that TRFs could be used to discriminate among fungal types. Sequence analysis indicated that important ECM fungi including Russulaceae, Thelephorales, and Tricholomataceae could be fingerprinted with TRFLP. In addition, a fixed proportion of the DNA extracted from each morphotype from the same core was used in a pooling experiment used to assess whether previously identified fungal species types could be distinguished from one another within reconstructed communities. Since some morphotypes share TRFs, dual analysis of ITS1 and ITS2 was necessary for accurate fingerprinting of fungal types. Approximately, 5.0±0.3 phylotypes were detected per core with TRFLP-corrected morphotyping as compared to 4.0±0.4 phylotypes using TRFLP on pooled community samples. TRFLP made on experimental sporocarp communities suggested that reduced ECM richness with TRFLP may be partly caused by differences in the amount of DNA available for PCR and primer bias. Nonetheless, TRFLP on pooled morphotypes accounted for more than 93% of colonized root tips. The method can be used to facilitate analysis of mycorrhizal communities using root tips collected from soil cores.     

Isolation of single-copy human genes from a library of yeast artificial chromosome clones

A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA.     

Type-restricted neutralization of molecular clones of human immunodeficiency virus

In a study of the immunologic significance of the genetic diversity present within single isolates of human immunodeficiency virus type 1 (HIV-1), the neutralization of viruses derived from molecular clones of the HIV-1 strain HTLV-IIIB by an extensive panel of sera was compared. Sera from HIV-1-infected patients and from goats immunized with polyacrylamide gel-purified HIV-1 envelope glycoprotein (gp120), native gp120, or gp120-derived recombinant peptides, showed marked heterogeneity in neutralizing activity against these closely related viruses. The change of a single amino acid residue in gp120 may account for such "clonal restriction" of neutralizing activity.     

Serum hepatitis antigen (SH): rapid detection by high voltage immunoelectroosmophoresis

An immunoelectroosmophoretic technique for rapid detection of the antigen (SH) associated with the serum hepatitis virus has been devised. The technique maintains the specificity characteristic of the Ouchterlony gel-diffusion method, yet detects in 1 to 2 hours one-tenth the amount of antigen required for gel diffusion. The test has immediate application to blood-banking practice since it permits the screening of such labile products as platelets and fresh whole blood, and the detection of antigen in additional serums negative by the Ouchterlony technique.