A polymerase chain reaction assay for the detection of Leptospira spp. in bovine semen.

A rapid and specific method for the detection of pathogenic Leptospira spp. in bovine semen using the polymerase chain reaction (PCR) is described. The primers used were derived from an EcoR1/BamH1 fragment that hybridized strongly to chromosomal DNA from the hardjobovis serovar. Three different extraction methods were evaluated in this study: phenol-chloroform extraction method, proteinase K (PK) in 1% SDS, followed by phenol-chloroform, and phenol-chloroform followed by 1% cetyltrimethylammonium bromide (CTAB). A PCR product of approximately 500 base pairs (bp) in length was obtained when DNA from pure Leptospira culture was used as a template for PCR, regardless of the DNA extraction method used. The product was consistent with that predicted from the gene sequence. However, in semen seeded in vitro, as well as in semen from infected bulls, a PCR product was obtained only when the leptospiral DNA was extracted from the specimen using the CTAB method. In contrast, other methods used for DNA extraction did not generate suitable templates for the PCR procedure. This is the first PCR protocol developed to detect Leptospira in bovine semen. The PCR protocol provided a direct and unequivocal demonstration that Leptospira can be detected in semen of infected animals. The CTAB method was also used successfully in detecting Leptospira in the urine of infected animals. The PCR procedure was shown to be more sensitive than either the fluorescent antibody test (FAT) or culture for detecting the organism in urine.     

Multiplex-PCR typing of high molecular weight glutenin alleles in wheat

In Australian commercial cultivars, each high molecular weight glutenin (Glu-1) homoeologous locus consists of one of two predominant alleles: Glu-A1a (subunit Ax1) or Glu-A1b (subunit Ax2*) at the GluA1 locus, Glu-B1b (Bx7 and By8 subunits) or Glu-B1i (Bx17 and By18 subunits) at the Glu-B1 locus, and Glu-D1d (Dx5 and Dy10 subunits) or Glu-D1a (Dx2 and Dy12 subunits) at the Glu-D1 locus. PCR-based assays have been developed in this study to discriminate between these common alleles at each locus. Primers specific for the Glu-A1 Ax2* gene give a single fragment of 1319 bp only in the presence of this gene. Primers targeting the Glu-B1 locus resulted in a co-dominant marker for which the Bx7 genotype produced two fragments (630 bp and 766 bp) and the Bx17 genotype a single fragment (669 bp). The third pair of primers was specific for the Dx5 gene and resulted in a single band of 478 bp. A multiplexed PCR assay was established which permitted the discrimination of the major HMW glutenins in a single PCR reaction and agarose gel assay. As the HMW glutenin composition of a wheat line is extremely important in determining the functional properties of wheat gluten, these markers are useful for the purposes of marker-assisted breeding. These markers may also be useful for the purpose of DNA-based identification of wheat varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Quantitative trait loci influencing grain protein content in tetraploid wheats

Seed storage protein content of durum wheat (Triticum turgidum var. durum) has an important effect on nutritional value and pasta-making characteristics. The objective of this study was to determine by association with genetic markers the number, chromosomal location, and magnitude of effect of quantitative trait loci (QTLs) controlling protein concentration in kernels. A set of 65 recombinant inbred lines (RIs) was developed by single seed descent from a cross between cultivated durum wheat cv. ‘Messapia’ (low protein content) and accession MG4343 of the wild tetraploid wheat var. dicoccoides (high protein content). This population was characterized for eight morphological, six storage protein, one isozyme and 124 RFLP loci. Field trials were conducted in one location in 1993 and two locations in 1994. QTLs were mapped by regression analysis on each marker locus for each location and for the average across environments. A total of six putative QTLs were located on chromosome arms 4BS, SAL, 6AS, 6BS and 7BS. The number and size of QTLs detected varied across environments. The marker with the highest r² value per QTL in each environment and across environments was chosen for a multiple linear regression analysis, which explained 49.2- 56.4% of the phenotypic variation for protein content. Only some of the markers were found to be negatively associated with plant grain yield and/or seed weight in one or two of the environments.     

A model system study of the inhibition of heterocyclic aromatic amine formation by organosulfur compounds

Organosulfur compounds and sodium bisulfite significantly inhibited (P < 0.05) heterocyclic aromatic amine (HAA) formation in model systems containing phenylalanine, creatinine, and glucose. There was, however, no inhibition by the same compounds in a model system containing only phenylalanine and creatinine. Diallyl disulfide (DAD) and dipropyl disulfide (DPD) concentrations in the model systems were significantly decreased (P < 0.05) after heating for 10 min at 180 degrees C. Only very low concentrations of sulfhydryl groups (4.19 and 4.00 micromol) were produced on heating DAD and DPD for 30 min. Reaction of glucose and DAD produced several sulfur-containing compounds. After 10 min of heating at 180 degrees C, HAA formation in the control model systems was increased significantly, and DAD was an effective inhibitor during this heating period. Tetrahydrothiophene-3-one (THT) and tetrahydrothiophene (THP); two products resulting from the interaction of glucose and DAD, had no direct influence on HAA formation in the model systems.     

Surfactant Enhanced Desorption of TNT from Soil

Surfactant enhanced desorption of 2,4,6-trinitrotoluene (TNT) from contaminated soils at a military site was investigated. Anionic (SDS and DOWFAX 8390), cationic (CTAC and CTAB), and nonionic (Tween 80 and Brij 35) surfactants were first tested at concentrations ranging from 0.1 to 1%. The anionic and nonionic surfactants were further tested at concentrations of up to 10%. Anionic surfactants, particularly SDS, provided the best desorption of TNT from the soil. There was not any increase in TNT desorption for both the nonionics and cationics at concentrations ranging between 0.1 to 1% and the extent of desorption was found to be lower than the TNT desorption only by water. The competition of the negatively charged soil surfaces for the positively charged cationics and the neutral nonionic surfactants may constitute the underlying reason. TNT was significantly desorbed when the concentrations of Tween 80, DOWFAX 80 and SDS were increased up to 10%.     

Spring Precipitation as a Predictor for Peak Standing Crop of Mixed-Grass Prairie

Ranchers and range managers need a decision support tool that provides a reasonably accurate prediction of forage growth potential early in the season to help users make destocking decisions. Erroneous stocking rate decisions can have dire economic and environmental consequences, particularly when forage production is low. Predictions must be based on information that is easily obtained and relevant to the particular range. Our goal was to evaluate monthly precipitation in spring months as a potential predictor of forage production compared to annual and growing-season precipitation. We analyzed the relationships between grazed and ungrazed peak standing crop (PSC) and precipitation using nonlinear regression and a plateau model, Akaike’s information criterion for model selection, and data from three locations: Streeter, North Dakota; Miles City, Montana; and Cheyenne, Wyoming. The plateau model included a linear segment, representing precipitation limiting production, and a plateau, an estimate of average production when precipitation is no longer the limiting factor. Both the response and predictor variables were rescaled so variability in production from average production was related to variability in precipitation from the long-term average. We found that grazing did not affect the relationship between PSC and precipitation, nor were annual or growing-season precipitation good predictor variables. The best predictor variable was total precipitation in April and May for Montana, May and June for North Dakota, and April, May, and June for Wyoming, with r² ranging from 0.74 to 0.79 for precipitation less than long-term average. These results indicate that spring precipitation provides useful information for destocking decisions and can potentially be used to develop a decision support tool, and the results will guide our choice of possible predictor models for the tool.     

Stomatal conductance and gross photosynthesis of potato (Solanum tuberosum L.) as influenced by irradiance,temperature, and growth stage

Summary Stomatal conductance (SC) and gross photosynthesis (P_G) of two potato clones were measured simultaneously with a dual-isotope porometer. Maximum P_G rates by Russet Burbank in the field were at intermediate levels early in the season, nearly doubled during mid-season, and declined during August. Maximum SC and P_G rates by Russet Burbank in the field occurred about 11h00, although leaf temperature did not reach its maximum unit¹ about 16h00. P_G rates by Russet Burbank in the glasshouse were maximum at about 24–30°C; SC was maximum by about 24°C and remained relatively constant at higher temperatures even though P_G declined. Clone A6948-4 had higher P_G rates in the field than Russet Burbank under all conditions and did not reach maximum levels until 1350 μE. The P_G response by Russet Burbank to varied irradiance in the glasshouse was similar to its response in the field.

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Zusammenfassung Die stomatale Leitf?higkeit (SC) und die Bruttophotosynthese (P_G) von zwei Kartoffelklonen (Solanum tuberosum L. — Russet Burbank und A6948-4) wurden gleichzeitig mit einem Dual-Isotop-Porometer gemessen. Die maximalen P_G-Raten von Russet Burbank im Feld waren anfangs Saison mittleren Grades, w?hrend der Saisonmitte fast doppelt und im Lauf des Augustes nahmen sie ab (Abb. 1). Die maximalen SC- und P_G-Raten von Russet Burbank im Feld wurden um etwa 11 Uhr vormittags erreicht, obwohl die Blattemperatur ihr Maximum erst ungef?hr um 4 Uhr nachmittags erzielte (Tabelle 1). Die P_G-Raten von Russet Burbank im Glashaus waren bei ca. 24–30°C maximal; das Maximum von SC war ungef?hr bei 24°C und blieb bei h?heren Temperaturen relativ konstant, selbst wenn P_G abnahm (Abb. 2). Offenbar kann die Abnahme von P_G nicht Aenderungen in SC zugeschrieben werden, sondern muss auf eine direkte Temperaturhemmung von P_G zurückgeführt werden. Die P_G-Reaktion von Russet Burbank auf unterschiedliche Einstrahlung im Glashaus war gleich wie ihre Reaktion im Feld. Im Gegensatz zu manchen C₃-Spezies nahm die SC von Russet Burbank mit steigender Einstrahlung von 400 auf 1700 μE m² s¹ zu (Abb. 3). Vergleichende SC-Messungen mit Kartoffeln müssen offenbar bei konstanten und bekannten Lichtstufen vorgenommen werden. Die SC- und P_G-Reaktionen auf die Einstrahlungsstufen (Abb. 3) deuten darauf hin, dass Russet-Burbank-Pflanzen einen überm?ssigen stomatalen Wasserverlust haben und dass SC unter Feldbedingungen vielleicht ohne jede Reduktion in P_G reduziert werden k?nnte. Der Klon A6948-4 hatte h?here P_G-Raten im Feld als Russet Burbank unter allen Bedingungen und erreichte die maximalen Grade nicht bis 1,350 μE (Abb. 4). Diese Vergleiche illustrieren die natürliche Variabilit?t, die in Kartoffelzüchtungsprogrammen vorhanden ist, und best?tigen den Wert des Screenings, um jene auszulesen, die als Elternmaterial für Kreuzungen am nützlichsten sind.

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Résumé La conductance des stomates (SC) et la photosynthèse brute (P_G) de deux clones de pommes de terre (Solanum tuberosum L. —Russet Burbank et A6948-4) ont été mesurées simultanément en utilisant un poromètre à isotope dual. Les intensités maxima de P_G pour Russet Burbank, au champ, se situaient à des niveaux intermédiaires en début de saison, étaient pratiquement doubles en milieu de saison, puis déclinaient en ao?t (fig. 1). SC et P_G atteignaient leur maxima au champ vers 11 heures du matin, alors que la température des feuilles ne parvenait à son maximum que vers 4 heures de l’après-midi (tableau 1). En serre, pour Russet Burbank, P_G était maximum vers 24–30°C; SC était maximum vers 24°C puis restait constant aux températures supérieures, tandis que P_G diminuait (fig. 2). Apparemment, la diminution de P_G ne pouvait être attribuée à des variations de SC mais devait être d?e à une inhibition directe de P_G par la température. Pour Russet Burbank, la réponse de PG en serre à une variation de l’irradiation était identique à la réponse au champ. Contrastant avec de nombreuses espèces C₃, chez Russet Burbank, SC a augmenté avec l’irradiation de 400 à 1700 μEm² sec¹ (fig. 3). Pour les pommes de terre, les mesures comparatives de SC doivent de toute évidence être effectuées à des niveaux constants et connus d’éclairement. Les réponses de SC et P_G aux différents niveaux d’irradiation (fig. 3). suggèrent que les plantes de Russet Burbank perdraient trop d’eau par les stomates et que l’on pourrait peut être réduire SC dans les conditions du champ sans réduire P_G d’aucune fa?on. Quelles que soient les conditions, le cone A6948-4 avait des niveaux de P_G plus élevés au champ que Russet Burbank et n’atteignait un maximum que pour 1,350 μE (fig. 4). Ces comparaisons illustrent la variabilité naturelle que l’on rencontre dans les programmes d’hybridation des pommes de terre, et qui établit la valeur discriminante pour choisir quels seront les matériels parentaux les plus utiles pour les croisements.

     

Response of tomato plants to saline water as affected by carbon dioxide supplementation. I. Growth,yield and fruit quality

Summary

Tomato plants (Lycopersicon esculentum (L.) Mill. cv. F144) were irrigated with low concentrations of mixed salts; the highest level (E.C. 7 dS m^–1) simulated conditions used to produce quality tomatoes in the Negev highlands. CO₂ enrichment (to 1200.mmol mol^–1, given during the daytime) increased plant growth at the early stage of development. However, later growth enhancement was maintained only when combined with salt stress. In the absence of CO₂ supplementation, overall growth decreased with salt (7 dS m^–1) to 58% and fresh biomass yields to 53% of the controls. However, under elevated CO₂ concentrations total plant dry biomass was not reduced by salt stress. CO₂ enrichment of plants grown with 7 dS m^–1 salt increased total fresh fruit yields by 48% and maintained fruit quality in terms of total soluble salts, glucose and acidity. Fruit ripening was about 10.d earlier under CO₂ enrichment, regardless of salinity treatment. It is suggested that a combined utilization of brackish water and CO₂ supplementation may enable the production of high-quality fruits without incurring all the inevitable loss in yields associated with salt treatment.     

Response of tomato plants to saline water as affected by carbon dioxide supplementation. II. Physiological responses

Summary

Photosynthesis of tomato plants (Lycopersicon esculentum (L.) Mill. cv. F144) was studied under conditions of CO₂ supplementation and salinity. The purpose of the study was to elucidate the mechanisms underlying the effects of salinity on the acclimation of tomato plants to CO₂ supplementation. Plants were grown under either low (355.mmol mol^–1) or elevated (1200.6.50 mmol mol^–1) CO₂ and were irrigated with low concentrations of mixed salts. The highest salinity level (E.C. 7 dS m–1) was that used to produce quality tomatoes in the Negev highlands, in Israel. During early development (three weeks after planting), the net photosynthetic rate of the leaves was much higher under elevated CO₂, and other than a slight decrease in quantum yield efficiency as measured by fluorescence (DF/F 9 ^m ), no signs of acclimation to high levels of CO₂ were apparent. Clear acclimation to high CO₂ concentration was evide t ten weeks after planting when the net photosynthetic rate, photosynthetic capacity, and carboxylation efficiency of leaves of non-salinized plants were strongly suppressed under elevated CO₂. This was accompanied by reductions in carboxylation efficiency, Rubisco activity and PSII quantum yield, and an increased accumulation of leaf soluble sugars. The reduction in photosynthetic capacity in the high CO₂ plants was less in plants grown at the highest salinity level. This was correlated with an increase in the PSII quantum yield parameters (F_v/F_m) and DF/F 9 m ) but not with Rubisco activity which was affected by the CO₂ treatments only. These results explain the effects of high CO₂ on yields in tomatoes grown at high levels of salt (Li et al., 1999).