Dairy manure N mineralization estimates from incubations and litterbags

A laboratory incubation trial and a field litterbag study were conducted to determine the rate and magnitude of mineralization of dairy manure N components in a south central Wisconsin silt loam. Dairy manure components (urine, feces, or bedding, each ¹⁵N-labeled and the other components left unlabeled) were incubated in soil at 11, 18, or 25°C. Samples were taken at 14, 21, 42, 84, and 168 days and analyzed for mineralized N ( and ) and ¹⁵N abundance in the inorganic and organic fraction (at day 168 only). In the field study, nylon mesh (38 μm) litterbags filled with ¹⁵N-labeled manure (2000) or unlabeled manure (2000 and 2002) were placed 7.5 cm below the surface and excavated at 7, 14, 21, 28, 35 (2000 only), 42, 56, 84, 98, and 126 days after burial and at corn (Zea mays L.) harvest, after 142 days in 2002 and 154 days in 2000. In the incubation study, 50−60% of applied urine N was mineralized showing the importance of this manure N component as a source of plant available N. About 14−19% of applied N was mineralized from the fecal and bedding components. In the litterbag experiment, approximately 70% of the dry mass and 67% of the N was mineralized from the litterbags with similar amounts measured using either labeled or unlabeled N. Rates of manure organic matter decomposition and N mineralization were best predicted using single exponential models for both years with most of the release occurring during the first 21 days.     

An empirical approach to target DNA quantification in environmental samples using real-time polymerase chain reactions

Recent advances in molecular techniques have allowed for the routine examination of nucleic acids in environmental samples. Although current methodologies are very sensitive, accurate target DNA quantification from environmental samples remains challenging. To facilitate high-throughput DNA quantification from environmental samples, we developed a novel DNA quantification method based on a non-linear curve-fitting approach to extract additional information from quantitative PCR amplification curves and used the fitted parameters to develop multiple regression standard equations for target DNA quantification. A 3-parameter sigmoidal function performed superior to a 4-parameter Weibull function for generating the multiple regression standard equations. In a verification experiment, target DNA was quantified in a series of ‘unknown’ samples in three soils using this approach and the results were compared to target DNA values determined using corrected and uncorrected Ct-based (threshold cycle) methods. For each method, the deviations from the expected target DNA content were determined. Results clearly showed that over all DNA concentrations, target DNA content determined by the non-linear curve-fitting method was more accurate and more precise than values predicted by all other methods. Analysis of variance conducted on the predicted DNA contents also revealed fewer statistical artifacts with the non-linear curve fitting method compared to the conventional Ct-based methods. The novel approach described here is accurate, inexpensive, and very amenable for automation and high-throughput applications.     

Quantitation of transgenic plant DNA in leachate water: real-time polymerase chain reaction analysis

Roundup Ready (RR) genetically modified (GM) corn and soybean comprise a large portion of the annual planted acreage of GM crops. Plant growth and subsequent plant decomposition introduce the recombinant DNA (rDNA) into the soil environment, where its fate has not been completely researched. Little is known of the temporal and spatial distribution of plant-derived rDNA in the soil environment and in situ transport of plant DNA by leachate water has not been studied before. The objectives of this study were to determine whether sufficient quantities of plant rDNA were released by roots during growth and early decomposition to be detected in water collected after percolating through a soil profile and to determine the influence of temperature on DNA persistence in the leachate water. Individual plants of RR corn and RR soybean were grown in modified cylinders in a growth room, and the cylinders were flushed with rain water weekly. Immediately after collection, the leachate was subjected to DNA purification followed by rDNA quantification using real-time Polymerase Chain Reaction (PCR) analysis. To test the effects of temperature on plant DNA persistence in leachate water, water samples were spiked with known quantities of RR soybean or RR corn genomic DNA and DNA persistence was examined at 5, 15, and 25 degrees C. Differences in the amounts and temporal distributions of root-derived rDNA were observed between corn and soybean plants. The results suggest that rainfall events may distribute plant DNA throughout the soil and into leachate water. Half-lives of plant DNA in leachate water ranged from 1.2 to 26.7 h, and persistence was greater at colder temperatures (5 and 15 degrees C).     

Forage quality of white clover (Trifolium repens L.) × Caucasian clover (T. ambiguum M. Bieb.) hybrids and their grass companion when grown over three harvest years

Interspecific hybrids between white clover (Trifolium repens L.) and Caucasian clover (Trifolium ambiguum M. Bieb.) have been developed to introgress the rhizomatous growth habit into white clover, to increase persistence and drought tolerance. The forage quality of T. repens, T. ambiguum and the backcross 1 (BC1) and backcross 2 (BC2) hybrids and companion grass, when grown in mixtures with an intermediate perennial ryegrass (Lolium perenne L.) under a cutting‐only management, was measured. In vitro dry‐matter digestibility (DMD), water‐soluble carbohydrate (WSC) and crude protein (CP) concentrations of the legume and grass fractions were measured throughout the growing season over three harvest years. Trifolium repens had a lower WSC but a higher CP concentration than the perennial ryegrass companion in all harvest years and at all cuts. The legume fractions from the BC1 and BC2 hybrid plots had a higher WSC and a lower CP concentration but an in vitro DMD value comparable with white clover throughout the growing season and in each harvest year. The grass fractions from the mixtures with the backcross hybrids had a higher WSC and a lower CP concentration than the grass fraction from the T. repens plots, in all harvest years and throughout the growing season. No difference in in vitro DMD between parental species and backcross hybrids was observed. The implications of these results for the development of these hybrids and animal performance are discussed.     

Populations of aphids on named varieties of Irish potatoes

Populations of the green peach aphid,Myzus persicae (Sulzer), and the potato aphid,Macrosiphum euphorbiae (Thomas), were observed on 27 named varieties of Irish potatoes in the Yakima Valley of Washington intermittently from 1947 through 1968. The Epicure, Great Scot, and B5141-6 varieties produced relatively fewer green peach aphids than other varieties tested; Bliss Triumph and Red Pontiac produced relatively fewer potato aphids. Except for Doebay Red, the red-skinned varieties, Bliss Triumph, Red McLure, Norland, and Earnly Rose, were extremely to moderately productive of the green peach aphid. No correlation was apparent between abundance of aphids and the specific gravity or yield of tubers.     

Chloroplast DNA variation in European potato cultivars

Summary The chloroplast DNA of 27 potato cultivars and 2 wild species has been examined by Southern blotting using probes representing the entire chloroplast genome of barley. 16% of the probe × enzyme combinations detected polymorphism between the potato cultivars. Cvs Maris Piper, Croft, Shelagh, Stormont Enterprise and Estima and the wild species could be distinguished from the other cultivars by their chloroplast DNA restriction patterns. These results are discussed in relation to cytoplasmic diversity and potato improvement.     

Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley

The effects on barley starch and grain properties of four starch synthesis mutations were studied during the introgression of the mutations from diverse backgrounds into an elite variety. The lys5f (ADPglucose transporter), wax (granule-bound starch synthase), isa1 (debranching enzyme isoamylase 1) and sex6 (starch synthase IIa) mutations were introgressed into NFC Tipple to give mutant and wild-type BC₂F₄ families with different genomic contributions of the donor parent. Comparison of starch and grain properties between the donor parents, the BC₂F₄ families and NFC Tipple allowed the effects of the mutations to be distinguished from genetic background effects. The wax and sex6 mutations had marked effects on starch properties regardless of genetic background. The sex6 mutation conditioned low grain weight and starch content, but the wax mutation did not. The lys5 mutation conditioned low grain weight and starch content, but exceptionally high β-glucan contents. The isa1 mutation promotes synthesis of soluble α-glucan (phytoglycogen). Its introgression into NFC Tipple increased grain weight and total α-glucan content relative to the donor parent, but reduced the ratio of phytoglycogen to starch. This study shows that introgression of mutations into a common, commercial background provides new insights that could not be gained from the donor parent.