Effect of the methodology on circulating peptides determination and consequences on net flux measurements across the gastrointestinal tract of sheep.

Two methodologies for the measurement of peptide amino acids (PAA) in blood were compared to evaluate their effects on the measurement of the net flux of peptides across the gastrointestinal tract of sheep. These methods consisted of a chemical deproteinization of blood samples with sulfosalicylic acid (1.6 M, 0.1 ml for 1 ml of sample) or perchloric acid (1 M, 1 ml for 1 ml of sample) followed by ultrafiltration through a 3,000-Da cut-off filter (SSA + UF3 kD) or gel filtration through a Sephadex G-15 column (1,500-Da cut-off filter; PCA + G-15), respectively, prior to PAA analysis. Peptide concentrations as determined by amino acid concentrations before and after hydrolysis of samples were slightly greater with the SSA + UF3 kD (991 microM) than with the PCA + G-15 (605 microM) methodology. However, both methodologies gave similar net portal-drained viscera flux data in sheep fed on alfalfa pellets with histidine as the only significant uptake of peptide amino acid.     

Occipito-atlanto-axial malformation with atlanto-axial subluxation in an ataxic calf

Occipito-atlanto-axial malformation with atlanto-axial subluxation was diagnosed radiographically in a 4-day-old Holstein calf with a short neck, symmetric ataxia, and tetraparesis, indicative of a cervical spinal cord lesion. Necropsy confirmed the clinical diagnosis and revealed ribs on C-7, with partial fusion of C-7 and T-1. Histologic examination revealed focal degeneration of the spinal cord in the second cervical segment at the level of the atlanto-axial joint and extensive secondary neuronal fiber degeneration possibly caused by spinal cord trauma that occurred in utero.     

Reaction to the cotton root-knot nematodes and the pollen and seed fertility of kenaf-roselle (Hibiscus cannabinus × H. sabdariffa) allohexaploids

Summary Allohexaploid (2n = 108) F₂ and F₃ interspecific hybrids ofHibiscus cannabinus × H. sabdariffa were intermediate in their resistance to the cotton root-knot nematode (Meloidogyne incognita acrita Chitwood 1949) between their moderately resistant, tetraploid parent, roselle (Hibiscus sabdariffa L.) and their susceptible, diploid parent, kenaf (H. cannabinus L.). Whether or not this resistance, as measured by the amount of root galling, is sufficient to be of practical importance is not known at present. However, results were encouraging enough to suggest further testing of the hybrids for agronomic potential, stability, fertility, and response to root-knot, and other parasitic nematodes that attack this potentially important paper-pulp crop.Cooperative investigations of the Crops Research Division, Agricultural Research Service, US Department of Agriculture, and the Texas Agricultural Experiment Station.     

Control of preharvest aflatoxin contamination in maize by pyramiding QTL involved in resistance to ear-feeding insects and invasion by Aspergillus spp.

Several resistance sources and resistance mechanisms to aflatoxin formation and corn earworm (Helicoverpa zea Boddie) damage to maize (Zea mays L.) have been identified. Based on this knowledge, experiments were initiated toward achievement of the following objectives: (1) to confirm earlier determinations on resistance traits of germplasm sources and to identify quantitative trait loci (QTL) associated with each of the traits, and (2) upon estimation of the degree of QTL effects on each trait, to generate a maize population, with chemical and physical resistance to Aspergillus spp. and ear-feeding insects, for inbred development. A 2-year field experiment to evaluate selected genotypes inoculated with A. flavus and infested with corn earworm revealed that significant variation exists among the genotypes for aflatoxin contamination and corn earworm damage. The protection of maize ears against aflatoxin contamination was primarily dependent on resistance to fungal infection and ear-feeding insects, and excellent husk coverage and tightness. A major QTL (p1) identified on chromosome 1S had effects of 54.0, 42.1, and 28.3% on the phenotypic variability for concentrations of silk maysin, 3′-methoxymaysin+apimaysin, and chlorogenic acid, respectively. Markers/QTLs for husk phenotypic traits and total aflatoxin concentrations have been determined, but more detailed mapping of these chromosomic regions will be necessary to locate precise markers/QTLs for husk traits and aflatoxin production. Realizing the complexity of the Aspergillus–aflatoxin-maize system and the factors affecting aflatoxin contamination, we are directing our program toward marker-assisted breeding to enhance or improve general genetic resistance to ear-feeding insects and invasion by Aspergillus spp.     

Genetic control over grain damage in a spring barley mapping population

A genetic map was constructed using DNA‐based markers in a barley mapping population derived from the cross ‘Tankard’בLivet’, that was developed to explore the genetic control over grain damage in spring barley cultivars. Quantitative trait loci (QTL) were located for husk skinning, gape between the lemma and palea and splitting of the fused pericarp/testa/aleurone tissues. The QTL accounted for 70% of the genetic variation in Split and 60% of the genetic variation in Gape and Skinning. The QTL were clustered on chromosomes 1H, 4H, 5H, 6H and 7H. QTL analysis indicates the possibility of transgressive segregation for grain splitting and so the breeding of lines with more extreme splitting. This is of concern to the malting industry as, without extensive phenotypic assessment, such lines could be commercialized, as was the case of Landlord, and put malting barley supplies at risk. These findings are discussed in relation to the genetic control over traits including grain length and width.     

Genetic and environmental effects on glucosinolate content and chemoprotective potency of broccoli

Broccoli is well recognized as a source of glucosinolates and their isothiocyanate breakdown products. Glucoraphanin is one of the most abundant glucosinolates present in broccoli and its cognate isothiocyanate is sulphoraphane, a potent inducer of mammalian detoxication (phase 2) enzyme activity and anti‐cancer agent. This study was designed to measure: glucosinolate levels in broccoli florets from an array of genotypes grown in several environments; the elevation of a key phase 2 enzyme, quinone reductase, in mammalian cells exposed to floret extracts; and total broccoli head content. There were significant environmental and genotype‐by‐environment effects on levels of glucoraphanin and quinone reductase induction potential of broccoli heads; however, the effect of genotype was greater than that of environmental factors. The relative rankings among genotypes for glucoraphanin and quinone reductase induction potential changed, when expressed on a per head basis, rather than on a concentration basis. Correlations of trait means in one environment vs. means from a second were stronger for glucoraphanin and quinone reductase induction potential on a per head basis than on a fresh weight concentration basis. Results of this study indicate that development of a broccoli phenotype with a dense head and a high concentration of glucoraphanin to deliver maximum chemoprotective potential (high enzyme induction potential/glucoraphanin content) is a feasible goal.