New method for detecting cellular transforming genes

Tumor induction in athymic nude mice can be used to detect dominant transforming genes in cellular DNA. Mouse NIH 3T3 cells freshly transfected with either cloned Moloney sarcoma proviral DNA or cellular DNA's derived from virally transformed cells induced tumors when injected into athymic nu/nu mice. Tumors were also induced by cells transfected with DNA from two tumor-derived and one chemically transformed human cell lines. The mouse tumors induced by human cell line DNA's contained human DNA sequences, and DNA derived from these tumors was capable of inducing both tumors and foci on subsequent transfection. Tumor induction in nude mice represents a useful new method for the detection and selection of cells transformed by cellular oncogenes.     

Marine lava cave fauna: composition, biogeography, and origins

An assemblage of endemic cavernicolous marine invertebrates, including taxa found on both sides of the Atlantic Ocean of great phylogenetic age or with affinities to deep sea organisms, inhabits the Jameos del Agua cave, a sea waterflooded Holocene lava tube cave on Lanzarote in the Canary Islands. This marine cave contains both relicts from Tethyan times, such as an apparently new crustacean family belonging to what had been the monotypic class Remipedia, and relicts of groups that are now common only in the deep sea as well as species that occur outside the cave.     

Impact of weather variables and season on sporulation of Phytophthora pluvialis and Phytophthora kernoviae

Phytophthora pluvialis and Phytophthora kernoviae are the causal agents of important needle diseases on Pinus radiata in New Zealand. Little is known about the epidemiology of the diseases, making the development of control strategies challenging. To investigate the seasonality and climatic drivers of sporulation, inoculum traps, consisting of pine fascicles floating on water in plastic containers, were exchanged fortnightly at five sites in P. radiata plantations between February 2012 and December 2014. Sections of needle baits were plated onto selective media and growth of Phytophthora pluvialis and P. kernoviae recorded. To explore the generalizability of these data, they were compared to detection data for both pathogens from the New Zealand Forest Health Database (NZFHDB). Further, equivalent analyses on infection of Rhododendron ponticum by P. kernoviae in Cornwall, UK allowed the comparison of the epidemiology of P. kernoviae across different host systems and environments. In New Zealand, inoculum of P. pluvialis and P. kernoviae was detected between January–December and March–November, respectively. Inoculum of both species peaked in abundance in late winter. The probability of detecting P. pluvialis and P. kernoviae was greater at lower temperatures, while the probability of detecting P. pluvialis also increased during periods of wet weather. Similar patterns were observed in NZFHDB data. However, the seasonal pattern of infection by P. kernoviae in the UK was the opposite of that seen for sporulation in New Zealand. Phytophthora kernoviae was likely limited by warmer and drier summers in New Zealand, but by colder winter weather in the UK. These results emphasize the importance of considering both environmental drivers and thresholds in improving our understanding of pathogen epidemiology.     

Comparing mating designs to restore seed production of interspecific hybrids between Trifolium repens (white clover) and Trifolium uniflorum

Interspecific hybrids between Trifolium uniflorum and cultivated white clover (Trifolium repens) have highly useful characteristics for temperate pastoral systems derived from both parent species. However, the early hybrids (F₁ and BC₁) also have unacceptably poor seed production for commerce. This study analysed the basis for the poor seed production and investigated breeding strategies for overcoming the problem. The BC₁F₁ generation produced lower‐than‐expected numbers of heads per plant and seeds per floret. Backcrossing of selected hybrids to white clover corrected these deficiencies and created new variation. Seed numbers were also returned to near target levels by recurrent selection within the BC₁ generation. Thus, it was possible to retain a theoretical average of 25% of T. uniflorum genome and still achieve high seed production per plant. The BC₁F₂ and BC₂F₁ generations produced high seed numbers per plant, along with reasonable variation. Both of these second‐generation hybrid forms have high reproductive potential and should be the focus for the selection of the desired combinations of agronomic and seed production traits.     

LED-Induced Chlorophyll Fluorescence Spectral Analysis for the Early Detection and Monitoring of Cadmium Toxicity in Maize Plants

Chlorophyll fluorescence spectral analysis permits detection, monitoring, and evaluation of abiotic stresses upon healthy plants using illumination of a light source in the UV?CVIS spectral range. This technique indirectly assesses the amount of physiological stress caused by photosynthetic damage, specifically damage to photosystem II, in plants. The objective of this study was to detect the toxicity of cadmium in maize plants via spectral analysis of chlorophyll fluorescence. The analysis is noninvasive and nondestructive and is used to follow the temporal evolution of changes in the chlorophyll content and physiological state of Zea mays L. seedlings under cadmium stress. Conventional techniques were also used to evaluate the dry matter production and Cd accumulation in plant leaves. Plants exhibited a notable reduction in dry matter production and chlorophyll levels with the administration of increasing doses of Cd in the nutrient solution. The fluorescence analysis was sensitive to changes caused by Cd in maize plants, detecting damage caused by different treatments before visual symptoms were observed. This technique has a practical application and produces rapid results that can be used in the evaluation of Cd-induced stress in plants and the detection of areas contaminated by this element.     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

Nitrous oxide fluxes and denitrification sensitivity to temperature in Irish pasture soils

Nitrous oxide (N₂O) emissions from grazed pastures constitute approximately 28% of total global anthropogenic N₂O emissions. The aims of this study were to investigate the effect of inorganic N fertilizer application on fluxes of N₂O, quantify the emission factors (EFs) for a sandy loam soil which is typical of large areas in Ireland and to investigate denitrification sensitivity to temperature. Nitrous oxide flux measurements from a cut and grazed pasture field for 1 year and denitrification laboratory incubation were carried out. The soil pH was 7.3 and had a mean organic C and N content at 0–20 cm of 44.1 and 4.4 g/kg dry weight, respectively. The highest observed peaks of N₂O fluxes of 67 and 38.7 g N₂O‐N per hectare per day were associated with times of application of inorganic N fertilizer. Annual fluxes of N₂O from control and fertilized treatments were 1 and 2.4 kg N₂O‐N per hectare, respectively. Approximately 63% of the annual flux was associated with N fertilizer application. Multiple regression analysis revealed that soil nitrate and the interaction between soil nitrate and soil water content were the main factors controlling N₂O flux from the soil. The derived EF of 0.83% was approximately 66% of the IPCC default EF value of 1.25% as used by the Irish EPA to estimate greenhouse gases (GHGs) in Ireland. The IPCC‐revised EF value is 0.9%. A highly significant exponential regression (r² = 0.98) was found between denitrification and incubation temperature. The calculated Q₁₀ ranged from 4.4 to 6.2 for a temperature range of 10–25 °C and the activation energy was 47 kJ/mol. Our results show that denitrification is very sensitive to increasing temperature, suggesting that future global warming could lead to a significant increase in soil denitrification and consequently N₂O fluxes from soils.     

Properties and functions of proton pumps in higher plants

The properties of the major H⁺-pumps in higher plant cells are described: these are the H⁺-ATPase of the plasma membrane and the H⁺-ATPase and the H⁺-pyrophosphatase of the tonoplast. They are distinguished by their structure, mechanism of action, kinetic properties, and location within the cell. The uses and value of isolated and purified membrane vesicles in transport studies are discussed. A particular example is given by the use of plasma membrane vesicles, purified by phase partition, to study proton/ sucrose co-transport in Ricinus cotyledons. Current evidence for the interference with these pumps by growth regulators, toxins and xenobiotics is outlined.     

The physical and chemical properties of the herbicide cinmethylin (SD 95481)

Cinmethylin (SD 95481), is a novel herbicide developed for the selective pre-emergence control of many annual grass weeds in a wide range of temperate and tropical crops. Representing new herbicide chemistry, cinmethylin is in the cineole family. Cinmethylin is a mobile colourless liquid with a boiling point of 313°C under an inert atmosphere at atmospheric pressure. It has a density of 1015 kg m^?3 and a viscosity of 70–90 mPa s, both at 20°C. It is miscible in all proportions with most organic solvents but has a low solubility, 63 mg litre^?1, in water. It has a vapour pressure of 10.2 mPa (20°C) and the vapour pressure/temperature relationship is given by log_e P(Pa)=28.9–9816/T (K). The n-octanol/water partition coefficient is 6850 and soil organic matter/water sorption coefficient (K_om) ranges between 165 and 235 over the three types of soil used in these studies. Cinmethylin is stable in water over the pH range 3–11. Solutions of cinmethylin in water or solvents are reasonably stable to sunlight, though thin films on a quartz surface photooxidise mainly to an ester within 24 h. This rate can be reduced by the addition of photostabilisers or by sorption onto soil surfaces. In an inert atmosphere cinmethylin is stable to high temperatures, though, in air, oxidation occurs at temperatures above 100°C to give the same product as by photodecay.