Storage protein changes during zygotic embryogenesis in interior spruce

The major storage proteins isolated from protein bodies of embryo tissues of interior spruce Picea glauca (Moench) Voss/Picea engelmanii Parry had apparent molecular weights of 41, 35, 33, 24 and 22 kD. Minor proteins of 30 and 27.5 kD were also observed. Based on their solubility characteristics, the 41 kD protein was identified as a water and buffer-soluble albumin, and the 35, 33, 24 and 22 kD proteins were characterized as buffer-insoluble, high salt-soluble globulins. Two-dimensional electrophoresis revealed each protein was composed of several isoelectric variants. Developmentally specific accumulation of storage proteins was observed during embryogenesis. The 41 kD protein only accumulated during the later stages of cotyledon maturation, whereas the other storage proteins began to accumulate during the early stages of embryo development. All storage proteins showed major accumulations during cotyledon maturation.     

Relationships between gas exchange adaptation of Sitka x interior spruce genotypes and ribosomal DNA markers

Adaptive physiological changes were investigated in seven populations of Sitka (Picea sitchensis (Bong.) Carr.) x interior spruce (P. glauca (Moench) Voss x P. engelmannii Parry ex Engelm.) spanning the Nass-Skeena transition zone in British Columbia, Canada. Each population was represented by an Si rDNA index that was calculated from the relative optical densities on a gel autoradiogram of five ribosomal DNA bands characteristic of Sitka spruce and interior spruce. This index estimates the proportion of the genome contributed by interior spruce. Physiological adaptations were assessed by gas exchange parameters measured under both well-watered and drought conditions. Under well-watered conditions, Sitka spruce populations had higher maximal photosynthesis at saturating light and ambient CO(2), higher quantum yield at the light compensation point, and higher dark respiration than interior spruce populations. Sitka spruce populations also reached maximal photosynthesis at lower photosynthetically active radiation and higher CO(2) concentrations, and had higher stomatal densities that resulted in lower stomatal limitations to photosynthesis than interior spruce populations. In contrast, interior spruce populations exhibited greater drought tolerance than Sitka spruce populations. Their gas exchange rates declined at a slower rate in response to drought. They maintained higher gas exchange rates in response to moderate to severe drought (predawn plant water potentials = -1.5 MPa), and their photosynthetic rates recovered faster when they were rewatered after exposure to drought. Comparison of the seven populations indicated that physiological parameters were significantly related to the Si rDNA index. An increase in Si rDNA index was associated with proportional changes in physiological measurements, suggesting that genetic interchange among species with contrasting ecological adaptations can enhance the environmental adaptation of natural populations.     

Relationship between nuclear DNA markers and physiological parameters in Sitka x interior spruce populations

Eight populations of Sitka spruce (Picea sitchensis (Bong.) Carr.) and interior spruce (Picea glauca (Moench) Voss x Picea engelmannii Parry ex. Engelm.) seedlings were sampled from a zone of Sitka-interior spruce introgression in British Columbia, Canada. Restriction fragment length polymorphisms of the nuclear ribosomal RNA genes (rDNA) were used to define species-specific hybridization patterns for the Sitka spruce and interior spruce populations. Hybridization was estimated from an index based on the relative abundance of polymorphic rDNA combining bands for each population. Sitka x interior hybrid seedlings had an index value for the relative abundance of interior spruce rDNA (Si-rDNA) ranging from 0.07 (Lower Nass; the most westerly collected source) to 0.95 (Bulkley Valley low-elevation seed orchard). During shoot elongation, osmotic potential at saturation (Psi(sat)) and turgor loss point (Psi(tlp)) increased, whereas total turgor (Psi(PTotal)) decreased. After bud set in the summer and throughout the fall, Psi(sat) and Psi(tlp) decreased, whereas Psi(PTotal) increased. At all times of year, populations with a higher Si-rDNA index had lower Psi(tlp) and Psi(sat) and higher Psi(PTotal) than populations with a lower Si-rDNA index. During the fall, Sitka x interior hybrid seedlings exhibited a seasonal decline in the temperature causing 50% needle electrolyte leakage (LT(50)) and in the critical temperature indicating the initial point of freezing injury. Seedlings with a higher Si-rDNA index had lower LT(50) and critical temperature values indicating greater freezing tolerance in the fall. Throughout most of the year, seedling population Si-rDNA index was related to the degree of drought and freezing tolerance.     

The Control of Reactive Oxygen Species Influences Porcine Oocyte In Vitro Maturation

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H₂O₂ or O₂?^‐ production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O₂?^‐ and H₂O₂ scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O₂?^‐ and H₂O₂ during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.     

Effects of Restored Stream Buffers on Water Quality in Non-tidal Streams in the Choptank River Basin

Restoration of riparian buffers is an important component of nutrient reduction strategies in the Chesapeake Bay watershed. In 1998, Maryland adopted a Conservation Reserve Enhancement Program (CREP), which provides financial incentives to take agricultural land out of production to plant streamside vegetation. Between 1998 and 2005, 1–30% of streamside vegetation (average?=?11%), was restored to forest or managed grass in 15 agriculturally dominated sub-basins in the Choptank River basin, a tributary of Chesapeake Bay. Pre-existing forested buffers represented 10–48% of the streamside (average?=?33%), for a total of 12–61% buffered streamsides (average?=?44%). Using multi-year water quality data collected before and after CREP implementation (1986, 2003–2006), we were unable to detect significant effects of CREP on baseflow nutrient concentrations based on the area of restored buffer, the percentage of restored streamside, or the percentage of total riparian buffer in the sub-basins (p?>?0.05). Although CREP increased the average buffered streamside from 33% in the 1990s to 44% by 2005, N and P concentrations have not changed or have increased in some streams over the last 20 years. Reductions may not have occurred for the following reasons: (1) buffer age, width, and connectivity (gaps) between buffers are also important to nutrient reductions; (2) agricultural nutrient inputs may have increased during this period; and (3) riparian buffer restoration was not extensive enough by 2005 to have measurable affects on the stream water quality in these sub-basins. Significant effects of CREP may yet be resolved as the current CREP buffers mature; however, water quality data through 2006 in the Choptank basin do not yet show any significant effects.     

Study of the duration and distribution of equine influenza virus subtype 2 (H3N8) antigens in experimentally infected ponies in vivo.

The purpose of this experiment was to study the duration and distribution of equine influenza virus in actively infected ponies over a 3 wk period. Pony foals (6-8 mo old) were infected experimentally by nebulizing equine influenza subtype-2 virus ultrasonically through a face mask. Successful infection was clinically apparent as each of the foals (n = 6) had a febrile response, a deep hacking cough and mucopurulent nasal discharge for 7 to 10 d. The virus was isolated from nasopharyngeal swabs of all the ponies 3 and 5 d after infection and all the ponies seroconverted to the virus. Samples were taken from the nasopharynx, mid-trachea and the mainstem bronchus with cytology brushes through an endoscope as well as from bronchoalveolar lavage fluid. On days 3 to 7 post-infection, ciliacytophtorea (the presence of cilia and ciliated plates separated from columnar epithelial cells) was recognized on routine cytological stain. Indirect immunoperoxidase staining utilizing polyclonal antibodies demonstrated viral antigen in intact and fragmented ciliated epithelial cells and in fragments of ciliated plates. The infected cells and cell fragments were particularly evident on days 3 and 5 post-infection in the nasopharynx, mid-trachea and mainstem bronchus and on days 3 to 7 post-infection in the bronchoalveolar lavage samples. On days 7 and 21 post-infection, viral antigen was identified in vacuoles of alveolar macrophage-like cells collected by bronchoalveolar lavage. It can be concluded from this study that equine influenza virus can infect not only the upper airways but also the bronchial epithelium and that viral antigen can persist up to 21 d post-infection.     

Effects of water and diet acidification with and without antibiotics on weanling pig growth and microbial shedding

Two 5-wk experiments were conducted to determine the effects of water and diet acidification with and without antibiotics on weanling pig growth performance and microbial shedding. In Exp. 1, 204 pigs (19.2 d of age) were used in a 3 x 2 factorial, with 3 dietary treatments fed with or without water acidification (2.58 mL/L of a propionic acid blend; KEM SAN, Kemin Americas, Des Moines, IA). Dietary treatments were: 1) control, 2) control + 55 ppm of carbadox (CB), and 3) dietary acid [DA; control + 0.4% organic acid-based blend (fumaric, lactate, citric, propionic, and benzoic acids; Kemin Americas)] on d 0 to 7 followed by 0.2% inorganic acid-based blend (phosphoric, fumaric, lactic, and citric acids; Kemin Americas) on d 7 to 34. In Exp. 2, 210 pigs (average 18.3 d of age) were fed 1 of 3 dietary treatments: 1) control, 2) control + 55 ppm of CB, and 3) control + 38.6 ppm of tiamulin + 441 ppm of chlortetracycline on d 0 to 7 followed by 110 ppm of chlortetracycline on d 7 to 35 (TC) with or without dietary acidification (same as Exp. 1) in a 3 x 2 factorial arrangement of treatments. For both experiments, the pigs were allotted based on genetics, sex, and initial BW [5.5 kg (Exp. 1) or 5.6 kg (Exp. 2)]. Pigs were housed at 6 or 7 (Exp. 1) and 7 (Exp. 2) pigs/pen. Treatments were fed in 3 phases: d 0 to 7, 7 to 21, and 21 to 35 (34 d, Exp. 1). Fecal grab samples were collected from 3 pigs/pen on d 6, 20, and 33 for measurement of pH and Escherichia coli. During phase 3 and overall in Exp. 1, pigs fed CB had greater (P < 0.001) ADG (overall ADG, 389 vs. 348, and 348 g/d, respectively), ADFI (P < 0.007, 608 vs. 559, and 554 g/d, respectively), and d 34 BW (P < 0.001, 18.8 vs. 17.3, and 17.3 kg, respectively) than pigs fed NC and DA. Phase 3 ADG was improved (P < 0.01) by water acidification across all diets. In Exp. 2, pigs fed CB and TC had greater ADG (P < 0.004; 315 and 303 vs. 270 g/d, respectively), ADFI (P < 0.01), and d 35 BW (P < 0.002; 16.7 and 16.2 vs. 15.1 kg, respectively) than pigs fed NC. There was a tendency (P < 0.08) for an improvement in ADG when DA was added to the NC or TC, but decreased ADG when DA was added to CB.