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.     

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.     

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.     

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.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

Auxin and ethylene regulation of diameter growth in trees

Recent studies on the phytohormonal regulation of seasonal cell-division activity in the cambium, primary-wall radial expansion of cambial derivatives, differentiation of xylem cells, and growth of the cortex in forest trees of the north temperate zone are reviewed. Indol-3-ylacetic acid (IAA, auxin) has been characterized by combined gas chromatography-mass spectrometry (GC-MS) in the cambial region of Abies balsamea, Pinus densiflora, Pinus sylvestris and Quercus robur. All of the evidence supports the hypothesis that developing leaves and extending shoots are primary sources of IAA. The rate of ethylene emanation varies among conifer species when adjoining phloem and cambial tissues are incubated in vitro. The cambium from young cuttings of Abies balsamea produces more ethylene than that from older cuttings. Ethylene production by seven-year-old Abies balsamea cambium is substantially increased in vitro when the tissue is provided with exogenous 1-aminocyclopropane-1-carboxylic acid and IAA. In response to elevated ethylene concentrations, cortex growth is accelerated in both hardwood and conifer seedlings. Ethrel (2-chloroethylphosphonic acid) increases ray size and ray-cell number and promotes traumatic resin-canal development in xylem. In Ulmus americana, endogenous ethylene concentrations are inversely correlated with cambial activity. Ethylene decreases vessel diameter in Acer negundo, Acer platanoides and Ulmus americana. Several studies suggest that ethylene has a role in regulating reaction-wood formation in both conifers and hardwoods.     

Expression profiles of immunity and reproductive genes during transition period in Holstein cattle

The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real‐time PCR to determine mRNA levels for the immune‐related genes TLR 2, 4, 6, 7 and β‐defensin 5 in addition to the reproduction‐related genes prolactin and IGF‐I. Results showed significant up‐regulation of pro‐inflammatory‐selected genes, TLR 4, 6 7 and β‐defensin 5 at the third‐week post‐calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up‐regulated at calving and 1 week after parturition and then down‐regulated again at second and third week. Prolactin and IGF‐I genes expression levels were significantly and gradually increased mainly post‐partum. This research highlights that the expression patterns of TLRs, BNBD5, PRL and IGF‐I could be biomarkers to follow up immune and reproductive status of dairy cow at peri‐parturient period to predict the most susceptible risk time for disease incidence and to build up management protocol.     

Vitrification with Glutamine Improves Maturation Rate of Vitrified / Warmed Immature Bovine Oocytes

The current study examined the protective effects of l ‐glutamine and cytochalasin B during vitrification of immature bovine oocytes. Oocyte vitrification solution (PBS supplemented with 10% FCS, 25% EG, 25% DMSO and 0.5 m trehalose) was the vitrification control. Treatments were the addition of 7 μg/ml cytochalasin B, 80 mm glutamine or both cytochalasin and glutaminine for 30 s. After warming, oocytes were matured in vitro for 24 h, fixed and stained with Hoechst (33342) for nuclear maturation evaluation. l ‐glutamine improved the vitrified/warmed immature bovine oocytes viability (32.8%), increasing the nuclear maturation rates compared to other treatments and the no treatment vitrified control (17.4%). There was, however, no effect of cytochalasin B on in vitro maturation (14.4%).     

Carbon content variation in boles of mature sugar maple and giant sequoia

At present, a carbon (C) content of 50% (w/w) in dry wood is widely accepted as a generic value; however, few wood C measurements have been reported. We used elemental analysis to investigate C content per unit of dry matter and observed that it varied both radially and vertically in boles of two old-growth tree species: sugar maple (Acer saccharum Marsh.) and giant sequoia (Sequoiadendron giganteum (Lindl.) Bucholz). In sugar maple there was considerable variation in tree ring widths among four radii for particular annual layers of xylem, revealing that the annual rate of C assimilation differs around the circumference and from the base of each tree to its top, but the observed variation in C content was unrelated to diameter growth rate and strongly related to the calendar year when the wood was formed. Carbon content in sugar maple wood increased in an approximately linear fashion, from < 50 to 51% from pith to cambium, at both the base and top of the boles. In giant sequoia, C was essentially constant at > 55% across many hundreds of years of heartwood, but it declined abruptly at the sapwood-heartwood boundary and remained lower in all sapwood samples, an indication that heartwood formation involves anabolic metabolism. Factors that may be responsible for the different C contents and trends with age between sugar maple and sequoia trees are considered. Tree-ring data from this study do not support some of the key assumptions made by dendrochronology.