Genetic transformation of Brassica

Tissue culture regeneration of plants from Brassica species can be achieved from various explant types. Their performance, however, varies with the genotype and age of the explant and is sensitive to medium supplements and Agrobacterium cocultivation. Most Brassica transformations were carried out using Agrobacterium and it was established that nopaline and agropine strains of A. tumefaciens and A. rhizogenes, respectively, are the most efficient. Application of virulence gene induction is ambiguous. Binary vectors are mostly applied in spite of indications that cointegrate vectors are more efficient. Cotransformation is successful with two different bacteria or with one bacterium carrying both plasmids, however, it has proved problematic to integrate the two transgenes into separate loci. Transformations with wild type rol genes develop hairy roots, from which transgenic plants can be regenerated. Vectorless transformation is feasible with microprojectile bombardment of microspore derived embryos being the most promising. Antibiotic markers providing resistance to kanamycin, hygromycin, and spectinomycin are applicable as selectable markers, while chloramphenicol is not suitable. Among herbicides, phosphinotricin gives good results while chlorsulfuron is unsuitable for in vitro selection. Major therapeutic antibiotics being successfully applied are carbenicillin, cefotaxime, amoxycillin, and ticarcillin. Standard methods for evaluating transgenic plants also have been applied to Brassica species. Some breeding objectives obtained through genetic transformation are presented.     

Heritability of inflorescence and floret traits in gerbera

Summary Heritability, repeatability, and components of variance were estimated for 68 morphological characters describing the inflorescence, scape, and florets of Gerbera hybrida. The heritability estimates were generally moderate to high for simple dimension characters and characters which were the mean of several measurements, low to moderate for ratio and composite characters, and low for enumeration characters and characters describing the intraplant variation of traits. The repeatability estimates were generally low for intraplant variation characters, moderate for simple ratio characters, and largest for composite and mean characters. For most of the characters, the variance among inflorescences from the same plant was the largest portion of the total phenotypic variance.     

Components of genetic variation for cut-flower yield in the Davis Population of Gerbera

Summary Genetic variances for cut-flower yield in Gerbera hybrida were partitioned into additive and dominance components. Mean additive variance was 37.3 with a standard error of 12.9. Mean dominance variance was 12.7 with a standard error of 32.6. The dominance ratio was 0.34. Narrow sense heritability, estimated from these components of variance, averaged 0.42 with a standard error of 0.13. Heritabilities estimated from parent-offspring regression averaged 0.25. Gain from selection was less then expected; realized heritabilities were approximately 0.20.     

The presence of inositol phosphates in gastric pig digesta is affected by time after feeding a nonfermented or fermented liquid wheat- and barley-based diet

The objective was to quantify the retention of digesta and evaluate the degradation of phytate or inositol hexakisphosphate (InsP(6)) and lower inositol phosphates (InsP?, InsP?, InsP?, and InsP?) in the stomach at different times after feeding pigs a fermented liquid diet with microbial phytase or a nonfermented diet with or without microbial phytase. Six barrows fitted with gastric cannulas were used. The experiment was a 3 × 3 Latin square with 3 pigs fed 3 diets during 3 wk in 2 replicates. Each experimental period lasted for 7 d, comprising 3 d of adaptation and 4 d of total collection of gastric digesta. For each pig, the digesta was collected once daily at 1, 2, 3, or 5 h after feeding the morning meal. A basal wheat- and barley-based diet was steam-pelleted at 90°C. The dietary treatments were a nonfermented basal diet (NF-BD), the NF-BD with microbial phytase (750 phytase units of phytase/kg, as-fed basis; NF-BD + phytase), and the NF-BD + phytase fermented for 17.5 h (F-BD + phytase). Gastric InsP?-P was not detected at all in pigs fed F-BD + phytase because of complete InsP? degradation during fermentation of the feed before feeding. Gastric InsP?-P decreased over time (P < 0.05) in pigs fed NF-BD and NF-BD + phytase. The decreases were 45, 54, 56, and 61 percentage points greater at 1, 2, 3, and 5 h, respectively, in pigs fed NF-BD + phytase compared with NF-BD. However, substantial amounts of InsP? still passed into the small intestine in pigs fed NF-BD + phytase, especially within the first hour (estimated to 17% of InsP?-P intake). The accumulation of lower inositol phosphates in gastric digesta was very small for all treatments and at all times because of a rapid and almost complete degradation. In conclusion, phytase addition to the nonfermented diet increased the degradation of gastric InsP?. However, considerable amounts of intact InsP? still passed into the small intestine because of a shortage of time for InsP? degradation in the stomach. Therefore, to increase the apparent digestibility of plant P in dry wheat- and barley-based diets, the development of phytases that can degrade InsP? effectively immediately after ingestion of the feed at an initial gastric pH from 6.5 to 5.0 is needed. Feeding F-BD + phytase compensated for the shortage of time because the InsP? degradation was completed during fermentation before feeding. The degradation of InsP? to InsP? is the bottleneck for plant P utilization in pigs because the degradation of the lower inositol phosphates is rapid and almost complete.     

Prevalence of gastrointestinal nematodes in growing pigs in Kabale District in Uganda

During the last 30 years, pig production in Uganda and neighbouring counties has increased markedly. Pigs are mainly kept as a source of income for small-scale farmers; however, the pig production is subject to several constraints, one of them being worm infections. A study was carried out in rural communities in Kabale District in the South Western part of Uganda in September and October 2007 in order to estimate the prevalence of gastrointestinal nematode parasites in pigs based on coprological examination. Fifty-six households were randomly selected and visited. Housing system and deworming history were recorded. Faeces was sampled from rectum of one to five pigs (age, 3–12 months) per household. A total of 106 pigs were examined coprologically of which 91% excreted nematode eggs. The following prevalences of nematode eggs were recorded: strongyles (89%), Ascaris suum (40%), Trichuris suis (17%) and spiruroid eggs (48%). On household level, rearing pigs on slatted floors in pens significantly reduced the faecal egg excretion of strongyle eggs with almost 80% (p = 0.010) and a significant interaction between floor type and anthelmintic treatment was found for spiruroids (p = 0.037). Fifteen T. suis egg positive pigs were selected for post-mortem examination of the gastrointestinal tract. The post-mortem examinations revealed that 93% pigs were infected with Oesophagostomum spp. (worm burden, min–max 10–2,180), 73% with A. suum (1–36), 67% with T. suis (6–58), and 20% with Hyostrongylus rubidus (worms not quantified). In general, nematode infections were widespread and polyparasitism common in pigs in Uganda. However, worm burdens were moderate which may be related to recent deworming or to the practice of rearing pigs on slatted floors in wooden elevated pens.     

Observations of variable inter-observer agreement for clinical evaluation of faecal consistency in grow-finishing pigs

Inter-observer agreement for assessment of faecal consistency in pigs was evaluated using a scoring system with 3 categories. In a pilot study, 3 observers performed an examination of faecal samples post-collection. The samples were obtained from pigs (12-13 weeks old) in 4 herds with a history of diarrhoea associated with Lawsonia intracellularis, Brachyspira spp. and/or Porcine Circovirus Type 2. Observer 1 examined all the faecal samples from the 4 herds. Observer 2 only examined the faecal samples from herds 1 and 2. Observer 3 only examined the faecal samples from herds 3 and 4. We observed a substantial agreement in faecal consistency scores between Observers 1 and 3 (kappa=0.64, 95% CI: 0.51-0.78). In contrast, only a fair agreement was observed between Observers 1 and 2 (kappa=0.24, 95% CI: 0.14-0.34). The variations in inter-observer agreement detected in the current study suggest that misclassification error can be a problem in studies assessing faecal consistency. Solutions may include developing a standardized system for scoring the consistency of pig faeces, calibration when more than one observer is involved in clinical studies and using a more objective measure of faecal consistency.     

Secondary renal tubular acidosis in a Hereford calf

A 3-month-old Hereford heifer calf was presented for lethargy. Blood gas analysis and plasma biochemical testing revealed severe metabolic acidosis, azotemia, hyponatremia, hyperchloremia, and normal anion gap. Results of a urinalysis were consistent with acute tubular necrosis with inadequate acidification of urine based on the degree of acidemia. Salmonella enterica serovar agona was cultured from both urine and feces. The calf was treated with intravenous polyionic fluids, bicarbonate, and antimicrobials. Acidosis and azotemia resolved, and 4 months following initial presentation the heifer was clinically normal.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Condensed tannins and flavonoids from the forage legume sulla (Hedysarum coronarium)

The condensed tannin concentrations and composition and the characterization of the phenolic constituents in the leaves of the forage legume sulla (Hedysarum coronarium), a biennial forage legume found in temperate agricultural regions, were studied. The colorimetric butanol-HCl assay was used for the quantitation of the seasonal condensed tannin concentrations in the leaves of sulla. Fractionation of extracts on Sephadex LH-20 using step elution with aqueous methanol, followed with aqueous acetone or gradient elution with water, aqueous methanol, and aqueous acetone, gave condensed tannin and flavonoid fractions. The chemical characteristics of the purified condensed tannin fractions were studied by acid-catalyzed degradation with benzyl mercaptan and electrospray ionization mass spectrometry (ESI-MS). Thiolysis revealed that epigallocatechin was the major extender unit (15-75%) while gallocatechin was the major terminal unit (50-66%), thus indicating the extractable sulla condensed tannin fraction as the prodelphinidin type. Condensed tannin oligomers to polymers obtained from Sephadex LH-20 gradient fractions ranged between 2.9 and 46 mDP. The homo- and heterogeneous oligomer ions in condensed tannin gradient fractions detected by ESI-MS ranged from 2 to 10 DP and are consistent with the values obtained by thiolysis (2.9-6.9 DP). Lower molecular weight phenolics, including flavonoids and phenolic acids, were characterized by liquid chromatography atmospheric pressure chemical ionization mass spectrometry (LC-APCI/MS) and ESI/MS/MS on a linear ion trap. The flavonoids extracted with aqueous acetone and methanol from sulla leaves and identified included kaempferol, rutin, quercetin-7-O-α-L-rhamnosyl-3-O-glucosylrhamnoside, quercetin-3-O-α-L-rhamnosyl-7-O-glucoside, kaempferol-3-O-β-D-glucoside-dirhamnoside, genistein-7-O-β-D-glucosyl-6″-O-malonate, formononetin-7-O-β-D-glucoside-6″-O-malonate, and afrormosin and the phenolic acid chlorogenic acid.     

Cutaneous asthenia in the dog. A report of two cases

Two cases of cutaneous asthenia in Danish dogs are described. Case 1 was a Weimeraner; case 2 a Golden Retriever. In both instances the dogs had acquired numerous lesions of the skin within the first half year of their lives. The wounds exhibited good healing but led to broad, thin and pliable scars. Both dogs had in several instances received attention to gaping wounds, characterized by little tendency of bleeding. The clinical examination reveiled that the skin was hyperextensible and possessed pendulous folds, particularly behind the elbows. The degree of hyperextensibility was evaluated by means of a skin extensibility index. Furthermore, it was possible to manipulate the digital cushion in an unphysiological manner. One of the dogs were euthanised, and in case 2 the clinical diagnosis was confirmed by histopathological examination. No conclusion could be drawn regarding the mode of inheritance in the two cases, but existing literature strongly suggests the existence of a genetic disposition and thus, elimination of the disease related gene from any breeding program is of great importance.