Electrophoretic Characterization of Quinoa Seed Proteins

Quinoa (Chenopodium quinoa Willd) is an important domesticated food crop of the Andean highlands with potential as an alternative crop elsewhere. Among its most attractive characteristics are the quantity and favorable amino acid balance of the seed proteins. The objective of this study was to characterize quinoa seed proteins by electrophoretic mobility, solubility fractionation, and genetic variability from a wide genetic base. Electrophoretic profiles of denatured albumin, globulin, prolamin, and glutelin solubility fractions demonstrated that quinoa seed polypeptides could be classified as either albumin or globulin with most predominant polypeptides in the globulin fraction. Insignificant amounts of protein were present in the prolamin fraction and all polypeptides in the glutelin fraction had identical electrophoretic mobilities to albumins and globulins. Three globulin polypeptides of 34.3, 35.6, and 36.2 kilodaltons in size were highly variable within and among the accessions examined and appear to be coded by at least two loci. Two-dimensional peptide mapping revealed that these three polypeptides were homologous. These highly variable markers could be used for identification and classification of germplasm and elucidation of systematics and genetic variability within the quinoa germplasm pool. All other major polypeptides were electrophoretically invariant among the accessions examined.     

Triploid and Hexaploid Regenerants from Hexaploid Timothy (Phleum pratense L.) via Anther Culture

Pollen embryos were obtained from eight genotypes and viable green plants were regenerated from four genotypes in an anther-culture experiment with 165 genotypes of Phleum pratense L. Formation of proembryos inside the cultured anthers during the first 10—12 days was significantly influenced by genotypes and by the type of nutrient media. Primary embryos developed into multiple secondary embryos before regeneration of plants. Among a total of 62 plants regenerated, only 13 were albinos. Of the green regenerants, 11 were triploid while 35 were hexaploid when DNA-content was measured by flow cytometry. Eight plants with a triploid DNA-content did possess the triploid chromosome number of 21. Triploid and hexaploid regenerants from two different parents showed simplified isozyme (GPI and PGD) banding patterns relative to that of their parents.     

Enhancement of RAPD Analysis by Restriction-endonuclease Digestion of Template DNA in Wheat

Random amplified polymorphic DNA (RAPD) analysis has proven to be an effective procedure for molecular marker applications in plant breeding, although non-specific amplification may limit its utility in some species. The objective of this study was to determine the effectiveness of restriction-endonuclease digestion of template DNA for elimination of non-specific amplification and generation of heritable RAPD markers. Restriction endonucleases digested wheat DNA to completion in amplification buffer, suggesting that the restriction endonuclease can be added directly to the reaction mixture prior to amplification. A 1-h 37°C step was programmed into the thermal cycler for restriction-endonuclease digestion which was followed immediately by amplification. Non-specific amplification was reduced and DNA marker patterns were altered in digested samples when compared to undigested samples. Genetic segregation of two polymorphic markers tested in F₅ inbred progeny fit expected 1:1 ratios. These results suggest that heritable DNA markers may be generated with reduction in non-specific amplification when restriction-endonuclease digestion of template DNA is conducted as part of the RAPD procedure.     

The effect of lupins as compared with peas and oats on the yield of the subsequent winter barley crop

New high yielding early maturing cultivars of lupins have been introduced in north-west Europe as grain protein crops in crop rotations. This paper reports on a comparative study of lupins with peas and oats, and of their effect on yield of subsequent winter barley crops. These crops were given five levels of N under irrigated and non-irrigated conditions on sand and loam. Under rain fed conditions the grain yield of pea, oat and lupin varied between 24–36, 34–53 and 18–37 hkg DM ha⁻¹, respectively. Supplemental irrigation raised grain yield of oat to 50–60 hkg DM ha⁻¹, while grain yield in pea was not affected and grain yield in lupin in most cases decreased due to gray mould attack and excessive vegetative growth in the indeterminate lupin variety. Under rain fed conditions, the grain nitrogen content of pea, oat and lupin varied between 137–172, 61–80 and 189–226 kg N ha⁻¹, respectively, and was significantly higher in lupin as compared with pea. On sandy soil, similar low-root densities were found for pea, oat and lupin below 30 cm depth. On sand, at final harvest the residual soil-N of lupin and pea, as measured in a subsequent winter barley crop not supplied with N fertilizer, was 15 and 8–10 kg N ha⁻¹ higher than in winter barley following oat, respectively. The nature of the probably more N-root residues of lupin is discussed. On loam, the residual N of lupin and pea was similar, 18–27 kg N ha⁻¹. On sand, under rain fed conditions preceding lupin and pea as compared with oat, increased the barley grain yield at zero N-application 77 and 49%, respectively; the effect of lupin was significantly higher than that of pea until the highest N-level 120 kg N-application ha⁻¹. On loam under rain fed conditions preceding lupin and pea increased the barley grain yield at zero N-application by 36 and 62%, respectively, as compared with oat; at N-application>60 kg N ha⁻¹ the grain yield was similar after all three crops. For both soil types the same level of effect was found under irrigated conditions. Conclusions: Supplemental irrigation might result in lower grain yield in lupin due to gray mould attack and excessive growth if indeterminate lupin varieties are used. Grain nitrogen yield of lupin is significantly higher than that of pea. On sand, the effect of lupin on the subsequent winter barley grain yield is significantly higher than that of pea, probably due to greater N-root nitrogen residues. On loam, lupin and pea have similar effects on the subsequent winter barley crop.     

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.     

Effect of genotype,environment and carbohydrate on anther culture response in head cabbage (Brassica oleracea L. convar. capitata (L.) Alef.)

Summary The effect of genotype, growing conditions for donor plants and type and concentration of carbohydrate in the culture medium was investigated for anther culture of head cabbage (white cabbage, savoy cabbage, pointed-headed cabbage). Strong genotypic effects on embryo formation from the cultured anthers were shown as well as superior embryo formation from anthers of field grown donor plants compared to plants grown in the greenhouse. When comparing 7, 10 and 13% sucrose in the medium, embryo response increased with increasing sucrose concentration. With maltose, which was generally inferior to sucrose as carbohydrate source for anther culture, the embryo response did not increase with maltose concentration above 10 per cent.     

Anther Culture Response in Perennial Ryegrass (Lolium perenne L.)

20 diploid clones from 7 varieties, and 10 tetraploid clones from 3 varieties of Lolium perenne were tested in replicated anther culture experiments. Embryos or calluses, were obtained from all clones, and plants were regenerated from all clones except one. The total yield of plants (albino and green plants) ranged from 0 to 61 plants per 100 cultured anthers among genotypes, and there was a general tendency for tetraploic genotypes to be more responsive. Viable green plants were obtained from 5 diploid and 7 tetraploid clones representing 2 and 3 varieties, respectively. Their origin from reduced pollen was confirmed by a haploid chromosome number in some regenerants and by homozygosity for isozyme loci in spontaneously chromosome doubled plants produced from heterozygous diploid donor plants. A large number of the plants were successfully established in the soil. For most donor genotypes, green plants were rare exceptions, but two diploic clones consistently produced 2.3 and 3.8 green plants per 100 cultured anthers, respectively. Estimates of variance components from replicates with greenhouse and field-grown donor plants showed that genotypes accounted for about 73 per cent of the total variation in yield of embryos/calluses, while only 14—15 per cent of the total variation was due to replicates. Hence at present, emphasis should be placed on die selection of high-response genotypes in material of high agronomic potential.     

Urban Influences on Stream Chemistry and Biology in the Big Brushy Creek Watershed, South Carolina

Naturally high total dissolved solids and upstream agricultural runoff often mask the influence of urban land cover on stream chemistry and biology. We examined the influence of headwater urbanization on the water chemistry, microbiology, and fish communities of the Big Brushy Creek watershed, a 96 km² drainage basin in the piedmont of South Carolina, USA. Concentrations of most major anions and cations (especially nitrate, sulfate, chloride, sodium, potassium, and calcium) were highest in the urban headwaters and decreased downstream. Generally, the highest concentrations of suspended coliform bacteria occurred in the urban headwaters. In contrast, stream habitat quality and the abundance, species richness, and species diversity of fishes did not differ significantly between urban and rural sites. Discharge of wastewater treatment plant effluent at one rural location caused an increase in concentrations of many solutes and possibly the abundance of benthic algae. We hypothesize that atmospheric dry deposition and domestic animal wastes are important sources of stream solutes and of coliform bacteria, respectively, in the urban headwaters. The lack of significant differences in fish abundance and diversity between urban and rural sites may indicate that urban development in the Big Brushy Creek watershed has not yet degraded habitat conditions greatly for stream fishes. Alternatively, agriculture or other land uses may have degraded stream habitat quality throughout the watershed prior to urbanization.     

The stability of soft drinks intake from adolescence to adult age and the association between long-term consumption of soft drinks and lifestyle factors and body weight

OBJECTIVES: To investigate the tracking of sugar-sweetened, carbonated soft drinks intake from age 15 to 33 years and the association between this intake and lifestyle factors and body weight. DESIGN: A longitudinal study with 18-20 years of follow-up. Data about diet, physical activity, smoking and dieting were collected in 1981/1979, 1991 and 1999. Body weight and height were measured in 1981/1979 and self-reported in 1999. SETTING: Oslo, Norway. SUBJECTS: Four hundred and twenty-two men and women. RESULTS: Tracking of soft drinks intake from adolescence into early adulthood (age 25 years) and from early adulthood into later adulthood (33 years) was moderate to high, while tracking from adolescence into later adulthood was low. Comparing those reporting a high intake of soft drinks in both 1991 and 1999 with those reporting a low intake at both times, male long-term high consumers were more likely to smoke (48 vs. 21%, P=0.002) and reported higher intakes of energy (12.2 vs. 10.2 MJ day(-1), P=0.005) and sugar (142 vs. 50 g day(-1), P<0.001) in 1999 than did long-term low consumers. Women high consumers were less likely to be physically active (14 vs. 42%, P=0.03) and had higher sugar intake (87 vs. 41 g day(-1), P<0.001) in 1999 than did women low consumers. There were no differences in body mass index, overweight or obesity in 1999 between long-term high and low consumers. CONCLUSION: In this study, stability of soft drinks intake from age 15 to 25 years and from age 25 to 33 years was moderate to high, while from age 15 to 33 years it was low. Soft drinks intake from age 25 to 33 years was associated with smoking and physical inactivity, but not with body weight.     

Breeding gains and changes in morphotype of Nordic spring wheat (1901–1993) under contrasting environments

Phenotypic diversity among 75 Nordic spring wheat cultivars was assessed in a glasshouse experiment, in which plots had no fertilizer or received 14-3-23 NPK plus Mg. On average, the fertilizer application delayed flowering by one day, shortened plant height (PH), as well as enhanced the number of fertile tillers (NFT), fresh and dry straw weight (FSW and DSW, respectively), but influenced negatively the dry matter content in the straw (DMCS) and the number of kernels per spike (K/S). The cultivar-by-fertilizer interaction did not affect significantly days to flowering (DF), PH, FSW, DSW, DMCS, thousand kernel weight, and K/S. Only NFT was significantly affected by this interaction. There were significant differences among cultivars for all characteristics, and the breeding gains were significant for improving earliness (as determined by DF), shortening PH, as well as for reducing DSW and DMCS irrespective of the environment. On average, FSW was low in newer cultivars grown in high inputs, while NFT was low in newer cultivars when grown in low inputs. The country of origin affected significantly DF, PH, DMCS, K/S, and 1000-kernel weight. On average, cultivars developed for Finland and Norway were significantly taller, and had higher DMCS. Finnish cultivars also flowered 1 or 2 days earlier and showed higher K/S than cultivars adopted in southern Scandinavia or Norway.