Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Analysis of dihaploid induction and production ability and seed parent x pollinator interaction in potato

Summary The production of dihaploids is the first step in a potato breeding program at the diploid level. Dihaploid induction ability, dihaploid production ability and seed parent x pollinator interaction were analyzed for 28 × 3, seed parent x pollinator combinations. This is the first report on significant interaction between seed parents and pollinators. Despite this interaction, IVP 101 had a significantly higher dihaploid induction ability than the widely used pollinators IVP 35 and IVP 48. Previous findings on significant differences in dihaploid production ability between seed parents were clearly confirmed.     

Somatic hybridization as a tool for tomato breeding

Protoplast fusion can be used to produce somatic hybrids of species that cannot be obtained by sexual hybridization. The possibility to introgress genes from Solanum species into the cultivated tomato species Lycopersicon esculentum, and to obtain novel cytoplasm-nucleus combinations (cybrids) was considered as an important strategy to extend the genetic variation available for tomato breeding. Somatic hybrids between L. esculentum and other Lycopersicon species, as well as between L. esculentum and Solanum or Nicotiana species, have been produced. Specific mutants, genotypes with antibiotic resistances, and metabolic inhibition by iodoacetate or iodoacetamide and irradiation were used for the selection of hybrids. In addition, the improvement of protoplast culture techniques and the use of the favourable tissue culture traits derived from species such as L. peruvianum, which have been introduced into tomato by classical breeding, allowed the efficient recovery of somatic hybrids. However, the occurrence of somatic incongruity in fusion combinations of L. esculentum and Solanum and even more in L. esculentum and Nicotiana, did not allow the production of true cybrids and/or fertile hybrids, indicating the importance of both cytoplasm-nucleus and nucleus-nucleus interactions in somatic incongruity. Another problem with fusions between distantly related species is the strongly reduced fertility of the hybrids and the very limited homoeologous recombination between chromosomes of the parental species. Partial genome transfer from donor to recipient through microprotoplast (+) protoplast fusion, and the production of monosomic or disomic chromosome addition lines, light overcome some of these problems. In symmetric somatic hybrids between L. esculentum and S. tuberosum the occurrence of limited somatic and meiotic recombination was demonstrated. Fertile progeny plants could be obtained, though at a low frequency, when embryo rescue was performed on a large scale after backcrossing hexaploid somatic tomato (+) potato hybrids with a tetraploid potato genotype. The potential value of genomic in situ hybridization (GISH) and RFLPs for the analysis of the genome/chromosome composition of the hybrids has been demonstrated for intergeneric somatic hybrids between Lycopersicon and Solanum.Abbreviations cpDNA chloroplast DNA - mtDNA mitochondrial DNA     

Adaptation of quinoa (Chenopodium quinoa) to Northern European agriculture: studies on developmental pattern

The inheritance of resistance to clubroot, caused by Plasmodiophora brassicae in Brassica oleracea was studied in the F₁, F₂and backcross progenies of three crosses between resistant and susceptible doubled haploid lines. Fresh weights of affected (CW) and healthy (HRW) parts of the root system were measured and R = ln((HRW+0.5)/(CW+0.5)) was used as a measure of resistance. R was shown to have a normally distributed error term with a constant standard deviation over the entire scale. Analysis of means indicated the presence of two complementary resistance genes in one of the crosses. In the other two crosses analysis of means also indicated two-locus interactions, in these cases reducing the cumulative effects of loci homozygous for resistance or for susceptibility alleles. The numbers of segregating resistance genes in these two crosses could not be determined. The presence of epistatic interactions precluded analysis of variance; therefore in this case the measurements were not more informative than the symptom grades analyzed in a previous study.R showed a high rank correlation (-0.85) with symptom grades assigned to the same plants. The results of genetic analyses of R were generally in agreement with those obtained earlier using symptom grades. Symptom grades can therefore be regarded as a reliable measure of symptom development, despite their more subjective nature.     

Race specific resistance against Phytophthora infestans in potato is controlled by more genetic factors than only R-genes

Summary For RFLP mapping of R-genes, determining resistance to specific races of Phytophthora infestans in tetraploid potato, it is necessary to develop well segregating populations at the 2x level. During mapping studies, evidence was obtained that more genetic factor(s) are involved in the expression of R-genes than conventionally believed. Two experiments are described in which such an additional genetic factor was suppressing or enhancing the expression of unknown R _nand R _ifactors. R _nand R _iappeared to be present in the investigated plant material, containing R4 and R10, or in one of the susceptible crossing parents. In a third experiment, the expression and the segregation of the well known R1 gene was influenced by an additional genetic factor. In that case there were indications for a dominant suppressor. This was established by the selection of susceptible plants carrying a RFLP allele of probe GP21 closely linked to R1. In three of the four F₁ populations, resulting from crosses between such susceptible plants and susceptible tester plants, resistnat progenies were found. The resistance appeared to be R1-specific. This clearly indicates that in three of the four investigated susceptible plants, the R1 gene was still present but not expressed.     

Cytological studies on adventitious shoots and minitubers of a monoploid potato clone

Summary A three step procedure for adventitious shoot regeneration on leaf explants of monoploid potato clone H7322 and a minituber induction procedure on stem segments have been described. Chromosome counts on 92 adventitious shoots showed that 85% of them had been polyploidized, i.e., 71% were diploid, 1% tetraploid, and 13% were mixoploid. Cytophotometric studies on nuclei of soil grown tubers of tetraploid cv Astarte, of 1x, 2x and 4x adventitious shoots of H7322, and of diploid H²578 showed in all cases polyploidization with prominent classes up to 8C and 16C. However, nuclei of pith cells of 5 weeks old minitubers which had developed on monoploid H7322 itself or on 1x adventitious shoots of H7322 showed predominantly 1C and 2C values. Pith cells of minitubers of monoploid H7322 were screened, after iodine staining, for the presence of variant cells containing reddish-brown staining (amylose-free) starch. In more than 75% of the investigated minitubers one or a few of such variant cells were found indicating that such a variation occurs in minitubers of monoploid potato and that this variant character is expressed in cells of vegetative storage organs like potato tubers.     

Heterogeneity of bacterial populations and pesticide degradation potentials in the unsaturated zone of loamy and sandy soils

Soil microbial processes play an important role in relation to pesticide pollution of groundwater, and may be strongly influenced by hydrological and geochemical properties. The consequences of such heterogeneous environments on bacterial biomass, enzymatic activities, carbon utilisation patterns, and pesticide mineralisation potentials in the unsaturated zone of a sandy loam and a coarse sandy soil profile were studied. In sandy loam soil profiles the number of bacteria decreased from 10⁹ cells g^-1 in the surface layers to about 10⁷ cells g^-1 at 1.5-5 m depth. Simultaneously, the hydrolysis of fluorescein diacetate and arylsulfatase activity decreased to below the detection limit at about 1.5 m depth, and carbon utilisation patterns showed that bacterial populations from surface soil were significantly different from those from 4 m depth. Bacterial biomass and activity in macropore soil tended to be slightly higher than in matrix soil, and the carbon utilisation patterns of bacterial populations extracted from macropore soil and from matrix soil seemed to be different. Maximally 3% of ¹⁴C-labelled mecoprop and isoproturon was mineralised in soil from the 1-1.5 m depth, and less than 1.5% was mineralised in soil from the 3.5-4 m depth. The macropore soil tended to have a higher degradation potential than the matrix soil. The total number of bacteria in the coarse sandy soil profile decreased from about 10⁸ in the plough layer to 10⁷ cells g^-1 at 0.4-2 m. The enzymatic activities and the degradation potentials of ¹⁴C-labelled mecoprop and isoproturon were significantly correlated (r² >0.79) and showed a distinct decrease at about 0.4 m. In addition to the depth variability, a horizontal heterogeneity in this soil was observed as horizons or compartments that differed in colour, i.e. with different chemical composition and concentrations of Fe and organic matter. Counts of viable bacteria and measurements of fluorescein diacetate hydrolysis and arylsulfatase activity confirmed a high variability of microbial biomass and activity in the sandy soil profile.     

Generation of spatially coherent light at extreme ultraviolet wavelengths

We present spatial coherence measurements of extreme ultraviolet (EUV) light generated through the process of high-harmonic up-conversion of a femtosecond laser. With a phase-matched hollow-fiber geometry, the generated beam was found to exhibit essentially full spatial coherence. The coherence of this laser-like EUV source was shown by recording Gabor holograms of small objects. This work demonstrates the capability to perform EUV holography with a tabletop experimental setup. Such an EUV source, with low divergence and high spatial coherence, can be used for experiments involving high-precision metrology, inspection of optical components for EUV lithography, and microscopy and holography with nanometer resolution. Furthermore, the short time duration of the EUV radiation (a few femtoseconds) will enable EUV microscopy and holography to be performed with ultrahigh time resolution.     

High-Resolution Imaging by Fourier Transform X-ray Holography

Fourier transform x-ray holography has been used to image gold test objects with submicrometer structure, resolving features as small as 60 nanometers. The hologram-recording instrument uses coherent 3.4-nanometer radiation from the soft x-ray undulator beamline X1A at the National Synchrotron Light Source. The specimen to be imaged is placed near the first-order focal spot produced by a Fresnel zone plate; the other orders, chiefly the zeroth, illuminate the specimen. The wave scattered by the specimen interferes with the spherical reference wave from the focal spot, forming a hologram with fringes of low spatial frequency. The hologram is recorded in digital form by a charge-coupled device camera, and the specimen image is obtained by numerical reconstruction.