Development and application of functional markers in maize

Summary Functional markers (FMs) are derived from polymorphic sites within genes causally involved in phenotypic trait variation (Andersen, J.R. & T. Lübberstedt, 2003. Trends Plant Sci 8: 554–560). FM development requires allele sequences of functionally characterized genes from which polymorphic, functional motifs affecting plant phenotype can be identified. In maize and other species with low levels of linkage disequilibrium, association studies have the potential to identify sequence motifs, such as a few nucleotides or insertions/deletions, affecting trait expression. In one of the pioneering studies, nine sequence motifs in the dwarf8 gene of maize were shown to be associated with variation for flowering time (Thornsberry, J.M., M.M. Goodman, J. Doebley, S. Kresovich, D. Nielsen & E.S. Buckler, 2001. Nat Genet 28: 286–289). Proof of sequence motif function can be obtained by comparing isogenic genotypes differing in single sequence motifs. At current, the most appropriate approach for this purpose in crops is targeting induced local lesions in genomes (TILLING) (McCallum, C.M., L. Comai, E.A. Greene & S. Henikoff, 2000. Nat Biotechnol 18: 455–457). In central Europe, maize is mainly grown as forage crop, with forage quality as major trait, which can be determined as proportion of digestible neutral detergent fiber (DNDF). Brown midrib gene knock out mutations have been shown to be beneficial for forage quality but disadvantageous for overall agronomic performance. Two brown midrib genes (bm1 and bm3) have been shown to be involved in monolignol biosynthesis. These two and additional lignin biosynthesis genes have been isolated based on sequence homology. Additional candidate genes putatively affecting forage quality have been identified by expression profiling using, e.g., isogenic bm lines. Furthermore, we identified an association between a polymorphism at the COMT locus and DNDF in a collection of European elite inbred lines.     

Treatment of tracheal collapse with an intraluminal stent in a miniature horse

A 7-month-old miniature horse was referred for respiratory distress. Tracheal collapse at the level of the thoracic inlet was diagnosed. An intraluminal nitinol stent was placed with endoscopic guidance. Respiratory function was restored immediately after stent placement. The main complication observed during a 14-month follow-up period was growth of granulation tissue through the stent, which was controlled satisfactorily by electrocautery performed during endoscopy with the horse standing. Treatment of tracheal collapse with an intraluminal stent is an effective, practical, and minimally invasive procedure in miniature and young equids and ponies.     

Assessment of adrenocortical activity by non-invasive measurement of faecal cortisol metabolites in dromedary camels (Camelus dromedarius)

The aim of this study was to determine whether glucocorticoid production could be monitored non-invasively in dromedary camels by measuring faecal cortisol metabolites (FCMs). Five Sudanese dromedaries, two males and three females, were injected with a synthetic adrenocorticotropic hormone (ACTH) analogue. Blood samples were collected pre- and post-ACTH injection. Faeces were sampled after spontaneous defecation for five consecutive days (2 days before and 3 days after ACTH injection). Baseline plasma cortisol values ranged from 0.6 to 10.8 ng/ml in males and from 1.1 to 16.6 ng/ml in females, while peak values after ACTH injection were 10.9–41.9 in males and 10–42.2 ng/ml in females. Peak blood cortisol values were reached between 1.5 and 2.0 h after ACTH injection. The concentration of FCMs increased after ACTH injection in the faeces of both sexes, although steroid levels peaked earlier in males [24 h; (286.7–2,559.7 ng/g faeces)] than in females [36–48 h; (1,182.6–5,169.1 ng/g faeces)], reflecting increases of 3.1–8.3- and 4.3–8-fold above baseline levels. To detect chromatographic patterns of immunoreactive FCMs, faecal samples with high FCM concentrations from both sexes were pooled and subjected to reverse phase high performance liquid chromatography (RP-HPLC). RP-HPLC analysis revealed sex differences in the polarity of FCMs, with females showing more polar FCMs than males. We concluded that stimulation of adrenocortical activity by ACTH injection resulted in a measurable increase in blood cortisol that was reliably paralleled by increases in FCM levels. Thus, measurement of FCMs is a powerful tool for monitoring the adrenocortical responses of dromedaries to stressors in field conditions.     

Survival and respiration of marbled rabbitfish (Siganus rivulatus) fingerlings at various oxygen tensions

Marbled rabbitfish, Siganus rivulatus, is an economically valuable herbivorous fish and a potential candidate for warmwater aquaculture. This study was carried out to: (1) assess the effect of various oxygen concentrations on survival and behaviour of S. rivulatus fingerlings and (2) investigate the response of S. rivulatus to hypoxia and determine its critical oxygen tension (P_crit). In the first experiment, groups of rabbitfish (15 fish per group) were maintained for 1 h in waters of various oxygen concentrations. They were then transferred to well‐aerated tanks and observed for 72 h. Survival was recorded, fish behaviour at low oxygen concentrations observed, and LC₅₀ after 1‐h hypoxia and 72‐h recovery evaluated. In the second experiment, a series of stop‐flow respirometry experiments were performed during which dissolved oxygen was allowed to drop to 0.5 mg L^?1 and respiration rate recorded at various oxygen concentrations. In the first experiment, all fish survived for 1 h at oxygen concentration of 1.44 mg L^?1 and greater, but started dying at oxygen concentrations below 0.65 mg L^?1 (16% survival). The LC₅₀ of S. rivulatus fingerlings was 0.6 mg L^?1. Results of the second experiment showed that S. rivulatus is an oxyregulator until P_crit (1.7 mg L^?1 O₂) is reached, becoming an oxyconformer below this concentration. Findings allow for a better understanding of environmental oxygen tolerances and minimum acceptable oxygen concentration in rabbitfish aquaculture.     

Anaesthetic efficacy of clove oil,benzocaine, 2‐phenoxyethanol and tricaine methanesulfonate in juvenile marbled spinefoot (Siganus rivulatus)

Anaesthetics are used in aquaculture and fisheries to facilitate routine procedures, such as capture, handling, transportation, tagging, grading and measurements that can often cause injury or induce physiological stress. Two experiments were performed to assess the efficacies of four anaesthetic agents, clove oil, benzocaine, 2‐phenoxyethanol and MS‐222 on juvenile marbled spinefoot rabbitfish (Siganus rivulatus). In the first experiment we tested the lowest effective doses that produced induction and recovery times in 3 min or less and 5 min or less respectively. Dosages were 70 mg L^?1 for clove oil, 60–70 mg L^?1 for benzocaine, 400 μL L^?1 for 2‐phenoxyethanol and 100–125 mg L^?1 for MS‐222. In the second experiment, we determined optimal concentrations of the four anaesthetics if they were to be used to transport rabbitfish fry. Anaesthetic concentrations suitable for handling and transport were: 10–15 mg L^?1 of MS‐222, 5–10 mg L^?1 of benzocaine, 5 mg L^?1 of clove oil and 50–100 μL L^?1 of 2‐phenoxyethanol. All anaesthetic agents are acceptable for use on S. rivulatus, however, 2‐phenoxyethanol, MS‐222 and clove oil appear to be more suitable than benzocaine. Further studies need to be conducted on effects of high and low doses of anaesthetic agents on physiology of marbled spinefoot.     

Synthesis and characterization of new polyamides containing symmetrical and unsymmetrical thiadiazole rings

Diamino derivatives of thiadiazole namely, 2,5-diamino-1,3,4-thiadiazole and 3,5-diamino-1,2,4-thiadiazole were synthesized and characterized. Polyamides were synthesized by condensation of the two diamino derivatives with diacide chlorides. The structures of polyamides were verified by elemental analysis, FTIR, ¹H-NMR, ¹³C-NMR and mass spectroscopy. The polyamides possess good solubility in aprotic organic solvents such as DMF, DMAC, DMSO and NMP at room temperature. Intrinsic viscosity measurements indicate that the polyamides synthesized have moderate molecular weights. The thermal stability of these polyamides in nitrogen atmosphere is relatively good, especially for those polyamides containing phenylene ring in the backbone. Compared to the structurally related Kevlar aramide, using 2,5-diamino-1,3,4-thiadiazole and 3,5-diamino-1,2,4-thiadiazole instead of p-phenylene diamine results in reducing the melting points of polyamides to below 350 °C.     

Comparison of the efficiency of A–PAGE and SDS–PAGE,ISSRs and RAPDs in resolving genetic relationships among <Emphasis Type="Italic">Triticum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species

Species that belong to the genus Triticum L. and the genetically related Aegilops L. genus are important genetic and economic resources because they have an evolutionary relationship with the two main agricultural crops T. aestivum (bread wheat) and T. durum (durum wheat). Therefore, it is important to understand the genetic relationships among the cultivated wheat species and their wild relatives. The latter have a great role in the improvement of cultivated wheat. Molecular markers are the best choice and most reliable means to study these relationships accurately. In this study, we compared the efficiency of the biochemical methods A–PAGE and SDS–PAGE on seed storage proteins and the molecular methods RAPDs and ISSRs to explore the genetic relationships among seven species of Triticum and 20 Aegilops species. Three phylogenetic trees obtained in this study were compared with available classifications and phylogenetic trees constructed earlier for these species. It was noted that the tree based on ISSRs data was the most congruent with those classification and trees. This may be attributed to the fact that ISSRs is more specific, and therefore more reliable. This study is the first to study genetic relationships among all species studied here using biochemical and molecular techniques.     

Oyster Crassostrea virginica Spat Settlement as it Relates to the Restoration of Fish River Reef in Mobile Bay, Alabama

Abstract.— Oyster spat settlement at four oyster reefs in Mobile Bay, Alabama, USA, was studied from August through October 1998 and May through mid November 1999. Spat collectors at the reefs were replaced every 2 wk and spat-set estimated as number of oysters per meter square per day. Water quality data at Fish River Reef was monitored using remote sensors. Spat-set data revealed significant variation between the four sites and between the 2 yr. Spat settlement was 5 to 10 times greater at the other three reefs than at Fish River Reef. Dates and intensity of oyster settlement at Fish River Reef were different from dates and intensity of oyster settlement at Shell Bank Reef, both on the eastern side of the bay. However, settlement was similar between Cedar Point Reef and White House Reef, both on the western side of the bay. Spat set appears to occur 3 wk after a rapid decline in water temperature, provided adequate oxygen concentrations are present at the time of settlement. Data collected suggest that intensity of settlement at Fish River Reef is considerably less than at other reefs in this study but could be adequate to reestablish the reef, if cultch and environmental conditions are suitable. The data also suggest that the source of larval oysters at Fish River Reef is different from the source of larval oysters at the other sites tested in the present study.