Virus infections reduce in vitro multiplication of ‘Malling Landmark’ raspberry

Summary Virus-infected plants are often symptomless and may be inadvertently used as explant sources in tissue culture research. Our objective was to determine the effect of virus infection on micropropagation. We studied the effects of single and multiple infections of three common raspberry viruses on the in vitro culture of ‘Malling Landmark’ red raspberry (Rubus idaeus L.). Virus-infected reaspberry plants were produced by leaf-graft inoculation from known-infected plants onto virus-free ‘Malling Landmark’. Single-virus source plants were infected with either tobacco streak ilarvirus (TSV), tomato ringspot nepovirus (TomRSV), or raspberry bushy dwarf idaeovirus (RBDV) and were free of other viruses as determined by enzyme-linked immunosorbent assay (ELISA) and bioassay. Virus-free, single, and multiple virus-infected ‘malling Landmark’ explants were initiated into culture and multiplied on Anderson's medium with 8.9 μM N⁶-benzyladenine (BA). At the end of the multiplication tests, ELISA reconfirmed virus infections. In vitro multiplication of ‘Malling Landmark’ was significantly reduced by multiple infections, and multiplication of plants infected with all three viruses (RBDV+TomRSV+TSV) was less than half that of virus free cultures. Shoot height and morphology of in vitro cultures were not influenced by virus infection. The greenhouse stock plant with the three-virus infection was stunted and yellow compared to the control and the other infected plants. Part of a thesis submitted by C.-W.V.T. in partial fulfilment of the requirements for the MS degree. The use of trade names in this publication does not imply endorsement by the U.S. Department of Agriculture or Oregon State University.     

Hybrid origin of some ornamentals of Allium subgenus Melanocrommyum verified with GISH and RAPD

 Random amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) methods have been used to verify the hybridogenic origin and to identify the parental species of some ornamental cultivars in the subgenus Melanocrommyum of the genus Allium. The cultivars had been selected from seed obtained after uncontrolled pollination in breeders’ fields. The combination of GISH analysis with RAPD markers is very suitable for testing the hybridogenic origin of plants and to ascertain the parental species of the hybrids in such cases. As suspected, A. macleanii and A. cristophii are the parental species of ‘Globemaster’. The parental species of cultivar ‘Globus’ are A. karataviense and A. stipitatum, and not A. cristophii and A. giganteum as has been assumed on morphological grounds. Cultivars ‘Lucy Ball’ and ‘Gladiator’ are of hybrid origin, though only one of the parental species, A. hollandicum, could be confirmed. The cultivars ‘Purple Sensation’, ‘Mount Everest’, ‘White Giant’, ‘Michael H. Hoog’ and ‘Mars’ are not hybrids since neither GISH nor RAPD suggest the presence of a second genome. ‘Purple Sensation’ belongs to A. hollandicum, ‘Mount Everest’, ‘White Giant’ and ‘Mars’ to A. stipitatum,‘Michael H. Hoog’ to A. rosenorum. Received: 3 July 1997 / Accepted: 9 October 1997     

Impact of sulfur on density of <Emphasis Type="Italic">Tetranychus pacificus</Emphasis> (Acari: Tetranychidae) and <Emphasis Type="Italic">Galendromus occidentalis</Emphasis> (Acari: Phytoseiidae) in a central California vineyard

Sulfur is the oldest and most widely used fungicide in the vineyards of California, where it is used for control of powdery mildew (Uncinula necator [Schw.] Burr). For decades, sulfur use has been associated with outbreaks of Tetranychus pacificus McGregor (Acari: Tetranychidae) on cultivated grapes in the San Joaquin Valley. I undertook large-scale field studies to test this association, to evaluate the impact of sulfur on Galendromus occidentalis (Nesbit) (Acari: Phytoseiidae), a major predator of T. pacificus, and to determine if timing of sulfur applications with respect to grape bloom has an impact on T. pacificus density. The studies took place in a 32 ha vineyard in Fresno County, and all fungicide applications were made with commercial-scale equipment. In 1998 a ‘high sulfur’ treatment, a combination of wettable sulfur and sulfur dust, was compared to ‘low sulfur,’ in which demethylation inhibitor (DMI) fungicides partially substituted for sulfur. In 1999 treatments were ‘sulfur,’ ‘DMI,’ ‘sulfur pre-bloom’ (here sulfur was applied prior to grape bloom, in late May, and then DMIs were applied until mid-season) and ‘sulfur post-bloom’ (the reverse of ‘sulfur pre-bloom’). In each year, the T. pacificus population increase came after the end of fungicide applications, and results clearly show a relationship between sulfur use and T. pacificus density. In 1998, mean T. pacificus density was 2.7 times higher and mean G. occidentalis density 2.5 times higher in ‘high sulfur’ compared to ‘low sulfur.’ In 1999, the highest T. pacificus counts were in the ‘sulfur’ and ‘sulfur pre-bloom’ treatments, 4.8 times higher than ‘sulfur post-bloom’ and 2 times higher than ‘DMIs.’ Density of G. occidentalis was 2.3 times as high in ‘sulfur’ or ‘sulfur pre-bloom’ than ‘DMIs.’ The predator/prey ratio was not significantly different among treatments in 1998, but in 1999 it was highest in the ‘sulfur pre-bloom’ treatment. In 1999, density of Homeopronematus anconai (Baker) (Acari: Tydeidae) was 2.7 times higher in ‘sulfur pre-bloom’ compared to ‘sulfur,’ and higher by 2.7 times in ‘DMI’ compared to ‘sulfur post-bloom,’ suggesting a negative effect of sulfur on this tydeid. These results do not support the hypotheses that the cause of the increase in T. pacificus density is due to negative effects of sulfur on phytoseiids or tydeids. Rather, it appears that a plant-based explanation is likely, first, because of the differences in pre-bloom versus post-bloom sulfuring, and second, because of the long lag time between the end of the sulfur applications and the corresponding increase in spider mite density.     

Plasmid profiles of bacteriocin-producingLactobacillus isolates from African fermented foods

Fourteen of 200Lactobacillus isolates from African fermented foods,viz. ‘wara’, ‘kenkey’, ‘ugba’, ‘ogi’, ‘kunuzarki’, ‘fufu’ and ‘iru’ were found to produce bacteriocins againstL. plantarum and only three bacteriocinogenic isolates inhibited some of the food pathogens. Plasmid analysis of the 14 bacteriocin-producing lactobacilli showed that only 5 isolates harbored plasmids ranging in size from 3.1 to 55.5 kb.     

pORE: a modular binary vector series suited for both monocot and dicot plant transformation

We present a series of 14 binary vectors suitable for Agrobacterium-mediated transformation of dicotyledonous plants and adaptable for biolistic transformation of monocotyledonous plants. The vector size has been minimized by eliminating all non-essential elements from the vector backbone and T-DNA regions while maintaining the ability to replicate independently. The smallest of the vector series is 6.3 kb and possesses an extensive multiple cloning site with 21 unique restriction endonuclease sites that are compatible with common cloning, protein expression, yeast two-hybrid and other binary vectors. The T-DNA region was engineered using a synthetic designer oligonucleotide resulting in an entirely modular system whereby any vector element can be independently exchanged. The high copy number ColE1 origin of replication has been included to enhance plasmid yield in Escherichia coli. FRT recombination sites flank the selectable marker cassette regions and allow for in planta excision by FLP recombinase. The pORE series consists of three basic types; an ‘open’ set for general plant transformation, a ‘reporter’ set for promoter analysis and an ‘expression’ set for constitutive expression of transgenes. The sets comprise various combinations of promoters (P _HPL, P _ENTCUP2 and P _TAPADH), selectable markers (nptII and pat) and reporter genes (gusA and smgfp).     

Resistance to Phytophthora fragariae var. fragariae in strawberry: the Rpf2 gene

  Phytophthora fragariae var. fragariae is the causal agent of red stele (red core) root rot in strawberry (Fragaria spp.). The inheritance of resistance to one isolate of this fungus was studied in 12 segregating populations of F.×ananassa derived from crosses between four resistant cultivars (‘Climax’, ‘Redgauntlet’, ‘Siletz’, and ‘Sparkle’) and three susceptible cultivars (‘Blakemore’, ‘Glasa’, and ‘Senga’ Sengana’). The analysis clearly supports the hypothesis of a single segregating dominant resistance gene. It is proposed that this gene be designated Rpf2. Received 12 November 1996 / Accepted: 22 November 1996     

Self-sterility in the mutant ‘Zigui shatian’ pummelo (<Emphasis Type="Italic">Citrus grandis</Emphasis> Osbeck) is due to abnormal post-zygotic embryo development and not self-incompatibility

‘Shatian’ pummelo (Citrus grandis Osbeck), one of the main citrus cultivars in China, is self-incompatible, and its pollen tubes are believed to be arrested in style after self-pollination.We have characterized one ‘Shatian’ pummelo mutant, named ‘Zigui shatian’ pummelo. The mutant pummelo had identical DNA ploidy level, morphology (leaf shape, stoma size and density, pollen shape and size) and developmental progress of pistil and male organs to that of the common ‘Shatian’ pummelo. However, unlike the common ‘Shatian’ pummelo, ‘Zigui shatian’ is self-compatible since its pollen tubes can self-pollinate allowing for successful fertilization. Histological analyses of ‘Shatian’ pummelo further verified abnormal post-zygotic development which led to seed abortion. Simple sequence repeats (SSR) analysis revealed polymorphism in 1 of the 120 primers screened showing that ‘Zigui shatian’ and ‘Shatian’ pummelo are different at the DNA level. Taken together, these data suggested mutant ‘Zigui shatian’ pummelo might be derived from ‘Shatian’ pummelo with self-sterility by self-incongruity after self-fertilization.     

Dynamic metabolomic data analysis: a tutorial review

In metabolomics, time-resolved, dynamic or temporal data is more and more collected. The number of methods to analyze such data, however, is very limited and in most cases the dynamic nature of the data is not even taken into account. This paper reviews current methods in use for analyzing dynamic metabolomic data. Moreover, some methods from other fields of science that may be of use to analyze such dynamic metabolomics data are described in some detail. The methods are put in a general framework after providing a formal definition on what constitutes a ‘dynamic’ method. Some of the methods are illustrated with real-life metabolomics examples.