Designing parallel specifications in CCS

Journal ArticleWe describe a style of specifying concurrent systems based upon the parallel composition operatorof CCS and apply it to several asynchronous hardware examples

Statewide survey for streptomycin-resistant fire blight in orchards

Streptomycin is the industry standard for managing fire blight on apples and pears. In 2002, streptomycin-resistance was confirmed in Wayne County and eradicated through prompt action. In October 2011, streptomycin-resistance was confirmed in four apple blocks in Wayne and Ontario Counties. We conducted a statewide survey in 2012 to locate and characterize any additional resistant bacterial populations. A total of 174 samples were collected. Erwinia amylovora was isolated from 129 of these and 15 of these isolates were resistant to streptomycin (11.6%). Preliminary phylogenetic results show that all streptomycin-resistant isolates carry the streptomycin resistance genes, strA and strB, on transposon Tn5393 which resides on the non-conjugative plasmid, pEa29, meaning that the isolates had to be spread physically. The survey found seven new locations in which resistant bacteria are present in Wayne, Ontario, Monroe, Orleans, and Niagara counties. These counties are now considered 'High Risk Areas' for streptomycin-resistant fire blight in 2013. To prepare for the 2013 growing season, we prepared new sample submission guidelines and fire blight best IPM practices for distribution by faculty and Extension educators, at the winter fruit schools, and at the Empire State Producers Ex

Activation of the Pathogen-Inducible Gst1 Promoter of Potato after Elicitation by Venturia inaequalis and Erwinia amylovora in Transgenic Apple ( Malus × Domestica )

Rather than using a constitutive promoter to drive transgenes for resistance against fungal and bacterial diseases in genetic engineering of apple (Malus × domestica) cultivars, a promoter induced only after infection was preferred. The ability of the Pgst1 promoter from potato (Solanum tuberosum L.) to drive expression of the gusA reporter gene was determined in two genotypes of apple: the fruit cultivar Royal Gala and the M.26 rootstock. β-glucuronidase activity in the transgenic lines grown in a growth chamber was determined quantitatively using fluorometric assays and compared to the activity in Cauliflower Mosaic Virus (CaMV) 35S promoter-driven transgenic lines. In both apple genotypes, the Pgst1 promoter exhibited a low level of expression after bacterial and fungal inoculation compared to the level obtained with the PCaMV35S promoter (15% and 8% respectively). The Pgst1 promoter was systematically activated in apple at the site of infection with a fungal pathogen. It was also activated after treatment with salicylic acid, but not after wounding. Taken together, these data show that, although the Pgst1 promoter is less active than the PCaMV35S promoter in apple, its pathogen responsiveness could be useful in driving the expression of transgenes to promote bacterial and fungal disease resistanc

Synergistic activity of endochitinase and exochitinase from Trichoderma atroviride (T. harzianum) against the pathogenic fungus (Venturia inaequalis) in transgenic apple plants

Genes from the biocontrol fungus Trichoderma atroviride encoding the antifungal proteins endochitinase or exochitinase (N-acetyl-β-D-hexosaminidase) were inserted into ‘Marshall McIntosh' apple singly and in combination. The genes were driven by a modified CaMV35S promoter. The resulting plants were screened for resistance to Venturia inaequalis, the causal agent of apple scab, and for effects of enzyme expression on growth. Disease resistance was correlated with the level of expression of either enzyme when expressed alone but exochitinase was less effective than endochitinase. The level of expression of endochitinase was negatively correlated with plant growth while exochitinase had no consistent effect on this character. Plants expressing both enzymes simultaneously were more resistant than plants expressing either single enzyme at the same level; analyses indicated that the two enzymes acted synergistically to reduce disease. Selected lines, especially one expressing low levels of endochitinase activity and moderate levels of exochitinase activity, were highly resistant in growth chamber trials and had negligible reduction in vigor relative to control plants. We believe that this is the first report of resistance in plants induced by expression of an N-acetylhexosaminidase and is the first report of in planta synergy between an exochitinase and an endochitinas

Stable expression and phenotypic impact of attacin E transgene in orchard grown apple trees over a 12 year period

<p>Abstract</p> <p>Background</p> <p>Transgenic trees currently are being produced by <it>Agrobacterium</it>-mediated transformation and biolistics. The future use of transformed trees on a commercial basis depends upon thorough evaluation of the potential environmental and public health risk of the modified plants, transgene stability over a prolonged period of time and the effect of the gene on tree and fruit characteristics. We studied the stability of expression and the effect on resistance to the fire blight disease of the lytic protein gene, <it>attacin E</it>, in the apple cultivar 'Galaxy' grown in the field for 12 years.</p> <p>Results</p> <p>Using Southern and western blot analysis, we compared transgene copy number and observed stability of expression of this gene in the leaves and fruit in several transformed lines during a 12 year period. No silenced transgenic plant was detected. Also the expression of this gene resulted in an increase in resistance to fire blight throughout 12 years of orchard trial and did not affect fruit shape, size, acidity, firmness, weight or sugar level, tree morphology, leaf shape or flower morphology or color compared to the control.</p> <p>Conclusion</p> <p>Overall, these results suggest that transgene expression in perennial species, such as fruit trees, remains stable in time and space, over extended periods and in different organs. This report shows that it is possible to improve a desirable trait in apple, such as the resistance to a pathogen, through genetic engineering, without adverse alteration of fruit characteristics and tree shape.</p

Genetic transformation of apple ( Malus x domestica ) without use of a selectable marker gene

Selectable marker genes are widely used for the efficient transformation of crop plants. In most cases, antibiotic or herbicide resistance marker genes are preferred because they tend to be most efficient. Due mainly to consumer and grower concerns, considerable effort is being put into developing strategies (site-specific recombination, homologous recombination, transposition, and cotransformation) to eliminate the marker gene from the nuclear or chloroplast genome after selection. For the commercialization of genetically transformed plants, use of a completely marker-free technology would be desirable, since there would be no involvement of antibiotic resistance genes or other marker genes with negative connotations for the public. With this goal in mind, a technique for apple transformation was developed without use of any selectable marker. Transformation of the apple genotype "M.26” with the constructs pPin2Att35SGUSintron and pPin2MpNPR1 was achieved. In different experiments, 22.0-25.4% of regenerants showed integration of the gene of interest. Southern analysis in some transformed lines confirmed the integration of one copy of the gene. Some of these transformed lines have been propagated and used to determine the uniformity of transformed tissues in the plantlets. The majority of the lines are uniformly transformed plants, although some lines are chimeric, as also occurs with the conventional transformation procedure using a selectable marker gene. A second genotype of apple, "Galaxy,” was also transformed with the same constructs, with a transformation efficiency of 13

EST contig-based SSR linkage maps for Malus × domestica cv Royal Gala and an apple scab resistant accession of M. sieversii , the progenitor species of domestic apple

Malus sieversii is a progenitor species of domestic apple M.×domestica. Using population "GMAL 4595” of 188 individuals derived from a cross of Royal Gala×PI 613988 (apple scab resistant, M. sieversii), 287 SSR (simple sequence repeats) loci were mapped. Of these SSRs, 80 are published anchors and 207 are newly developed EST (expressed sequence tag) contig-based SSRs, representing 1,630 Malus EST accessions in GenBank. Putative gene functions of these EST contigs are diverse, including regulating plant growth, development and response to environmental stresses. Among the 80 published SSRs, 18 are PI 613988 specific, 38 are common and 24 are Royal Gala specific. Out of the 207 newly developed EST contig-based SSRs, 79 are PI 613988 specific, 45 are common and 83 are Royal Gala specific. These results led to the construction of a M. sieversii map (1,387.0cM) of 180 SSR markers and a Royal Gala map (1,283.4cM) of 190 SSR markers. Mapping of scab resistance was independently conducted in two subsets of population "GMAL 4595” that were inoculated with Ventura inaequalis races (1) and (2), respectively. In combination with the two major resistance reactions Chl (chlorotic lesions) and SN (stellate necrosis) to each race, four subsets of resistance data, i.e., Chl/race (1), SN/race (1), Chl/race (2) and SN/race (2), were constituted and analyzed, leading to four resistance loci mapped to the linkage group 2 of PI 613988; SNR1 (stellate necrosis resistance to race (1)) and SNR2 are tightly linked in a region of known scab resistance genes, and ChlR1 (Chlorotic lesion resistance to race (1)) and ChlR2 are also linked tightly but in a region without known scab resistance genes. The utility of the two linkage maps, the new EST contig-based markers and M. sieversii as sources of apple scab resistance are discusse

Foraging on human-derived foods by urban bird species

Capsule: Providing peanuts on bird feeders was shown to attract more individuals and more species than providing cheese or bread. Aims: To investigate how the provision of different human-derived foods affected visit rates of urban birds at bird feeders. Methods: A fully replicated study design was set up in parkland, offering a binary choice from three food types (peanuts, bread and cheese) on bird tables. Birds were observed by using a scan-sample method. Results: Peanuts attracted more visits, and a greater diversity of bird species, than cheese or bread. This preference was strongest for Blue Tits Cyanistes caeruleus and Great Tits Parus major, whereas European Robins Erithacus rubecula visited all food types equally, and Blackbirds Turdus merula preferred cheese. Bread was the most consumed food type when measured by mass, but this could be linked to varying bite sizes. Conclusion: Our results indicate that birds preferred to visit feeding stations with the most protein- and energy-rich foods, but that some birds still chose the carbohydrate-rich bread. The findings indicate that peanuts, rather than household scraps like bread and cheese, attract the highest number of species and individuals to bird tables. The findings will be of interest to the public and to organizations providing information on bird feeding for recreational purposes