Novel pGreen/pSoup dual-binary vector system in multiple T-DNA co-cultivation as a method of producing marker-free (clean gene) transgenic rice (Oriza sativa L) plant

The possibility of producing marker-free transgenic rice plants using a novel dual binary pGreen/pSoup vectors, in multiple T-DNA co-cultivation, was investigated and demonstrated to be feasible. The T-DNA in pSoup (pRT47) vector was engineered to contain the selection marker hygromycin phosphotransferase (aphIV) gene (plus intron in 5′ UTR), and the green fluorescent protein (gfp) as a reporter gene both driven by the CaMV35S promoter and the nopaline synthase terminator. T-DNA in the pGreen (pRT18) vector harboured the phosphinothricin acetyl transferase (bar), as selection marker gene, and the b- glucuronidase (gusA) plus intron as a reporter gene, both driven by the maize 5′ ubiquitin region and the nopaline synthase terminator. Both the pGreen and pSoup plasmids were transformed into E. coli strain DH5α using the PEG-transformation technique and into Agrobacterium strains AGL1using a freeze-thaw method. AGL1 was then used to transform embryogenic nodular units (ENU), derived from mature seeds of the model rice genotype Nipponbare. Selection on herbicide (PPT) or antibiotic (hygromycin) of co-cultured ENUs led to the production of numerous independently transformed callus clones containing both T-DNAs from the selected and unselected vector. While co- ransformation frequencies were 71% and 80% for the hygromycin only and herbicide (PPT) only selection, respectively, data showed that co-expression frequency is most useful for the production of marker free transgenic rice. About half (50%) of the independent transgenic plant lines contained at least one unlinked T-DNA integration. In this work, we showed for the first time, that the novel dual-binary pGreen/pSoup can efficiently produce marker-free transgenic rice.African Journal of Biotechnology Vol. 4 (6), pp. 531-540, 200

Chromosomal location of genes encoding for resistance to septoria tritici blotch (Mycosphaerella graminicola) in substitution lines of wheat

Chromosomal location of resistance to Mycosphaerella graminicola was studied in substitution lines of resistant Triticum genotypes into the (susceptible) cultivar Chinese Spring (T. aestivum). (Moderately) resistant genotypes for which substitution lines were available were tested in a first screening. We selected a synthetic hexaploid wheat (Synthetic 6x; T. dicoccoides × T. tauschii), T. spelta and the wheat (T. aestivum) cultivars Cheyenne and Cappelle-Desprez. In a second screening the most suitable Argentinian isolates were identified. We decided to use the isolate IPO 92067 (all sets of substitution lines), IPO 93014 (substitution lines of Synthetic 6x, Cappelle-Desprez and T. spelta) and IPO 92064 (substitution lines of Cheyenne). In the final experiments, substitution lines of the selected genotypes into Chinese Spring were grown in two different environments and inoculated with the selected isolates at the seedling stage (lines of all four selected genotypes) or the adult stage (lines of Synthetic 6x and Cheyenne). Resistance was expressed as (reduction in) necrosis percentage or pycnidial coverage percentage; the two measures were highly (linearly) correlated. When tested in the seedling stage, all chromosomes seemed to carry genes effective against M. graminicola. Many genes were effective against only one isolate or in only one environment or their effects only showed in one resistance parameter. Often these effects were minor. Only chromosome 7D of Synthetic 6x was found with a major effect against both isolates tested. When tested in the adult stage, all lines but the one carrying chromosome 4B from the resistant parent seemed to show genes effective against M. graminicola. The line carrying chromosome 7D from Synthetic 6x showed a level of resistance similar to the resistant parent for isolate IPO 92067, but not for isolate IPO 93014. Major genes, effective against both isolates, were also found on chromosomes 5A and 5D from Synthetic 6x. Lines carrying chromosome 1B, 5D or 6D from Cheyenne showed major effects against isolate IPO 92064. For both necrosis percentage and pycnidial coverage percentage, highly significant linear correlations were found between resistance in the seedling stage and resistance in the adult stage. However, the variance accounted for was only small (20-24%; n = 184).Facultad de Ciencias Agrarias y Forestale

Vascular plant changes in extreme environments: effects of multiple drivers

The Antarctic Peninsula is one of three regions of the planet that have experienced the highest rates of climate warming over recent decades. Based on a comprehensive large-scale resurvey, allowing comparison of new (2009) and historical data (1960s), we show that the two native Antarctic vascular plant species have exhibited significant increases in number of occupied sites and percent cover since the 1960s: Deschampsia antarctica increasing in coverage by 191 % and in number of sites by 104 %. Colobanthus quitensis increasing in coverage by 208 % and number of sites by 35 %. These changes likely occurred in response to increases of 1.2 °C in summer air temperature over the same time period. Both species exhibited changes with elevation due to the interaction of multiple drivers (climatic factors and animal disturbance), producing heterogeneity of responses across an elevation gradient. Below an elevation of 20 m fur seal activity exerted negative impacts. Between 20 and 60 m, both plant species underwent considerable increases in the number of sites and percent cover, likely influenced by both climate warming and nutrient input from seals. Above an elevation threshold of 60 m the maximum elevation of the sites occupied decreased for both species, perhaps as a consequence of physical disturbance at higher elevations due to the permafrost conditions and/or the snow cover thickness and persistence. Understanding the role of disturbance drivers for vegetation change in cold regions may become a research priority to enable improved forecasting of biological responses and feedbacks of climate warming on ecosystems in these globally influential regions

Chromosomal location of genes encoding for resistance to septoria tritici blotch (Mycosphaerella graminicola) in substitution lines of wheat

Chromosomal location of resistance to Mycosphaerella graminicola was studied in substitution lines of resistant Triticum genotypes into the (susceptible) cultivar Chinese Spring (T. aestivum). (Moderately) resistant genotypes for which substitution lines were available were tested in a first screening. We selected a synthetic hexaploid wheat (Synthetic 6x; T. dicoccoides × T. tauschii), T. spelta and the wheat (T. aestivum) cultivars Cheyenne and Cappelle-Desprez. In a second screening the most suitable Argentinian isolates were identified. We decided to use the isolate IPO 92067 (all sets of substitution lines), IPO 93014 (substitution lines of Synthetic 6x, Cappelle-Desprez and T. spelta) and IPO 92064 (substitution lines of Cheyenne). In the final experiments, substitution lines of the selected genotypes into Chinese Spring were grown in two different environments and inoculated with the selected isolates at the seedling stage (lines of all four selected genotypes) or the adult stage (lines of Synthetic 6x and Cheyenne). Resistance was expressed as (reduction in) necrosis percentage or pycnidial coverage percentage; the two measures were highly (linearly) correlated. When tested in the seedling stage, all chromosomes seemed to carry genes effective against M. graminicola. Many genes were effective against only one isolate or in only one environment or their effects only showed in one resistance parameter. Often these effects were minor. Only chromosome 7D of Synthetic 6x was found with a major effect against both isolates tested. When tested in the adult stage, all lines but the one carrying chromosome 4B from the resistant parent seemed to show genes effective against M. graminicola. The line carrying chromosome 7D from Synthetic 6x showed a level of resistance similar to the resistant parent for isolate IPO 92067, but not for isolate IPO 93014. Major genes, effective against both isolates, were also found on chromosomes 5A and 5D from Synthetic 6x. Lines carrying chromosome 1B, 5D or 6D from Cheyenne showed major effects against isolate IPO 92064. For both necrosis percentage and pycnidial coverage percentage, highly significant linear correlations were found between resistance in the seedling stage and resistance in the adult stage. However, the variance accounted for was only small (20-24%; n = 184).Facultad de Ciencias Agrarias y Forestale

Full sphere hydrodynamic and dynamo benchmarks

Convection in planetary cores can generate fluid flow and magnetic fields, and a number of sophisticated codes exist to simulate the dynamic behaviour of such systems. We report on the first community activity to compare numerical results of computer codes designed to calculate fluid flow within a whole sphere. The flows are incompressible and rapidly rotating and the forcing of the flow is either due to thermal convection or due to moving boundaries. All problems defined have solutions that allow easy comparison, since they are either steady, slowly drifting or perfectly periodic. The first two benchmarks are defined based on uniform internal heating within the sphere under the Boussinesq approximation with boundary conditions that are uniform in temperature and stress-free for the flow. Benchmark 1 is purely hydrodynamic, and has a drifting solution. Benchmark 2 is a magnetohydrodynamic benchmark that can generate oscillatory, purely periodic, flows and magnetic fields. In contrast, Benchmark 3 is a hydrodynamic rotating bubble benchmark using no slip boundary conditions that has a stationary solution. Results from a variety of types of code are reported, including codes that are fully spectral (based on spherical harmonic expansions in angular coordinates and polynomial expansions in radius), mixed spectral and finite difference, finite volume, finite element and also a mixed Fourier–finite element code. There is good agreement between codes. It is found that in Benchmarks 1 and 2, the approximation of a whole sphere problem by a domain that is a spherical shell (a sphere possessing an inner core) does not represent an adequate approximation to the system, since the results differ from whole sphere results

Copy Number Variation Affecting the Photoperiod-B1 and Vernalization-A1 Genes Is Associated with Altered Flowering Time in Wheat (Triticum aestivum)

The timing of flowering during the year is an important adaptive character affecting reproductive success in plants and is critical to crop yield. Flowering time has been extensively manipulated in crops such as wheat (Triticum aestivum L.) during domestication, and this enables them to grow productively in a wide range of environments. Several major genes controlling flowering time have been identified in wheat with mutant alleles having sequence changes such as insertions, deletions or point mutations. We investigated genetic variants in commercial varieties of wheat that regulate flowering by altering photoperiod response (Ppd-B1 alleles) or vernalization requirement (Vrn-A1 alleles) and for which no candidate mutation was found within the gene sequence. Genetic and genomic approaches showed that in both cases alleles conferring altered flowering time had an increased copy number of the gene and altered gene expression. Alleles with an increased copy number of Ppd-B1 confer an early flowering day neutral phenotype and have arisen independently at least twice. Plants with an increased copy number of Vrn-A1 have an increased requirement for vernalization so that longer periods of cold are required to potentiate flowering. The results suggest that copy number variation (CNV) plays a significant role in wheat adaptation

Inhibitory effects of climate change on the growth and extracellular enzyme activities of a widespread Antarctic soil fungus

Temperatures approaching or exceeding 20 °C have been measured during summer in polar regions at the surfaces of barren fellfield soils under cloudless skies around solar noon. However, despite the upper temperature limit for the growth of cold‐adapted microbes – which are abundant in polar soils and have pivotal roles in nutrient cycling – typically being close to this temperature, previous studies have not addressed the consequences of climate change for the metabolism of these organisms in the natural environment. Here, in a five‐year field experiment on Alexander Island in the southern maritime Antarctic, we show that the abundance of Pseudogymnoascus roseus, the most widespread decomposer fungus in maritime Antarctic fellfield soils, is reduced by 1–2 orders of magnitude when irrigated and nutrient‐amended soils are warmed to >20 °C during summer. Laboratory experiments under conditions mimicking those during midsummer in the natural environment indicated that the hyphal extension rates of P. roseus isolates and the activities of five extracellular enzymes are reduced by 54–96% at high water availability after exposure to temperatures cycling daily from 2–21 °C and 2–24 °C, relative to temperatures cycling from 2–18 °C. Given that the temperatures of surface soils at the study site already reach 19 °C during midsummer, the observations reported here suggest that, at predicted rates of warming arising from moderate greenhouse gas emissions, inhibitory effects of climate change on the metabolism of P. roseus could manifest themselves within the next few decades. Furthermore, with peak temperatures at the surfaces of fellfield soils at other maritime Antarctic locations and in High Arctic and alpine regions already exceeding 20 °C during summer, the observations suggest that climate warming has the potential to inhibit the growth of other cold‐adapted microbes, with negative effects on soils as the Earth’s climate continues to war