Development of colchicine‐induced tetraploid St. Augustinegrass (Stenotaphrum secundatum) lines

St. Augustinegrass is well suited for lawns and commercial landscapes. While many genotypes are cross‐fertile, all cultivars are propagated vegetatively in sod production. To ensure varietal purity, development of sterile triploid hybrids by crossing tetraploid and diploid genotypes has been successfully used in other warm‐season turfgrasses. Applying this model in St. Augustinegrass would be beneficial to sod producers and turf managers who require purity for certification and uniformity for performance, respectively. This study was conducted to develop colchicine‐induced tetraploid lines of St. Augustinegrass. Seeds of cultivar ‘Raleigh’ were treated with four colchicine concentrations at four exposure times. A non‐treated control was included among the treatments. Seedlings that germinated were screened for genome size changes using flow cytometry. Line DSA 13005 and two progeny lines derived through selfing, DSA 16001 and DSA 16016, were corroborated as tetraploids (2n = 4x = 36) through chromosome counts. These lines will be used in future breeding efforts to attempt development of sterile triploid cultivars of St. Augustinegrass.     

Assessing freeze-tolerance in St. Augustinegrass: temperature response and evaluation methods

Winter hardiness is a major-limiting factor for St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] grown in the transitional climatic region of the United States. Lab-based freeze tests that mimic the range of field winter survivability in St. Augustinegrass can contribute to the selection of cold hardy genotypes. This study used a whole container method, four freezing temperatures, and two data collection systems to evaluate the freezing response of nine St. Augustinegrass genotypes ranging in their winter hardiness. Results indicated ?3 and ?4 °C with average regrowth ratings of 33.6 and 17.8% respectively, were more suitable temperatures for evaluating freeze survival in St. Augustinegrass than ?5 and ?6 °C with average regrowth ratings of 0.4 and 0%, respectively. Visual ratings of surviving green tissue and regrowth were generally well correlated when evaluated over a six week period post-freeze with Pearson correlation coefficients ranging of 0.17–0.62  for ?3 °C freeze tests and 0.79–0.93 for ?4 °C freeze tests. Additionally, measurement of percent green cover using digital imaging techniques commonly utilized in turfgrass field studies were significantly correlated (0.66) with visual ratings averaged across weekly post-freeze evaluation measurements for both ?3 and ?4 °C freezing temperatures. These results provide evidence that digital imaging analyses are useful in estimating surviving green tissue and regrowth in lab-based freeze tests. This study provides additional information regarding freezing temperatures, genotype responses, and data collection methods in St. Augustinegrass, which should aid breeders in the improvement of freeze tolerance in the species.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Effects of gamma radiation on the immature stages ofLiriomyza Trifolii

The effects of γ-radiation on the immature stages ofLiriomyza trifolii Burgess were studied with doses of 2000-25 Gy. The high doses were lethal to all stages. At doses of 750 Gy or less, larvae survived to pupate, but their number was reduced and the majority were either dead or deformed and did not give rise to adults. Eggs and prepupae were more susceptible to radiation than other stages. Larval radiosensitivity decreased with progressing instar, and pupal sensitivity decreased with increasing age. Contribution from the Agricultural Research Organization. No. 2922-E, 1990 series.     

Characterizing the complexity of landscape boundaries by remote sensing

This paper presents a method for characterizing the complexity of landscape boundaries by remote sensing. This characterization is supported by a new boundary typology, that takes into account points where three or more landcovers converge (i.e., convergency points or coverts). Landscape boundary richness and diversity indices were proposed and calculated over 19 landscapes in South-East Brazil. Results showed that landscape boundaries, especially convergency points, provided an enrichment in landscape pattern analysis. Landcover boundary diversities were significantly related to landcover shape: elongated riparian units had the highest values for boundary diversity and coverts proportion indices. On the other hand, landscape analysis showed that indices of shape, richness, diversity and coverts proportion provided an additional evaluation of landcover spatial distribution within the landscape.     

Gene discovery in Eimeria tenella by immunoscreening cDNA expression libraries of sporozoites and schizonts with chicken intestinal antibodies

Specific antibodies were produced ex vivo from intestinal culture of Eimeria tenella infected chickens. The specificity of these intestinal antibodies was tested against different parasite stages. These antibodies were used to immunoscreen first generation schizont and sporozoite cDNA libraries permitting the identification of new E. tenella antigens. We obtained a total of 119 cDNA clones which were subjected to sequence analysis. The sequences coding for the proteins inducing local immune responses were compared with nucleotide or protein databases and with expressed sequence tags (ESTs) databases. We identified new Eimeria genes coding for heat shock proteins, a ribosomal protein, a pyruvate kinase and a pyridoxine kinase. Specific features of other sequences are discussed.     

Down-regulation of GPx1 mRNA and the loss of GPx1 activity causes cellular damage in the liver of selenium-deficient rabbits

The effects of 10 weeks of dietary selenium and/or vitamin E deficiency (< 0.03 mg Se and 1.5 mg vitamin E per kg diet) on body Se and vitamin E stores and on the down-regulation of liver cellular glutathione peroxidase (GPx1) and plasma glutathione peroxidase (GPx3) were examined in growing female New Zealand White rabbits in comparison to Se (+ 0.40 mg Se/kg diet) and/or vitamin E (+ 150 I.U./kg diet) supplemented controls. Additionally plasma lactate dehydrogenase (LDH) activity, liver thiobarbituric acid-reactive substances (TBA-RS) and liver protein carbonyls were measured to assess the development of oxidative stress during an alimentary Se and/or vitamin E deficiency. Significantly decreased concentrations of Se and vitamin E in plasma (Se: − 70%; vitamin E: − 87%) and liver (Se: − 90%; vitamin E: − 95%) indicated an efficacious Se and vitamin E depletion of the rabbits within 10 weeks. GPx1 messenger RNA levels (GPx1 mRNA) in the livers of Se-depleted rabbits were down-regulated to 1/3–1/8 of the Se supplemented controls. GPx1 enzyme activity in the livers of Se-deficient rabbits declined to 10% of the Se-supplied control rabbits. A significantly elevated LDH activity in the blood plasma of Se- and vitamin E-deficient rabbits indicated a general impairment of tissues. Markedly increased TBA-RS concentrations and protein carbonyl contents in the livers of Se- and vitamin E-deficient rabbits gave further evidence for severe oxidative damage of cellular lipids and proteins during an alimentary Se and/or vitamin E deficiency. Both a full expresssion of GPx1 attained by dietary Se supplementation and dietary vitamin E supply effected an almost complete protection against oxidative cellular damage of the liver.