Identificação de espécies de Fusicoccum causadoras de podridão em frutos de abacate

ABSTRACT The Fusicoccum genus of fungi are known to cause stem-end rot in various fruit plants, such as mango, guava, peach and avocado. Several species of this fungus are reported attacking avocado (Persea americana) in several countries. Based on this information, the present study aimed to identify species of Fusicoccum associated with rot in avocado fruits in the State of São Paulo. Samples were collected (fruits with rot symptoms) from regions of Bauru, Bernadino de Campos and Piraju. All isolates obtained had its pathogenicity confirmed by inoculation of healthy avocado fruits. After confirming its pathogenicity, these isolates had their DNA extracted and the ITS-5.8S rDNA region was amplified. After editing, these sequences were used to search for similar sequences in the NCBI. Eleven samples were identified as Neofusicoccum parvumand others were identified as Botryosphaeria dothidea(F. aesculi). Both species were found in all regions of collection.

Venäjän informaatio-psykologinen sodankäyntitapa terrorismin torjunnassa ja viiden päivän sodassa

This study examines Russia’s Pattern of Information-Psychologic Warfare in Counter-terrorism and in the Five Day War. The first case of this comparative case study research examines an internal national security crisis, namely two of the most notorious terrorist strikes that took place firstly in The Moscow Dubrovka Theater in October 2002 and two years later in Beslan School 2004 in September. In the second case an external national security crisis, the Five Day War conducted in August 2008 between the Russian Federation and the state of Georgia will be analyzed. At the centre of this research report lays an idea: a war of information by using information as the target and as a weapon. Based on a comparative case study setting this study tries to understand how Russian pattern of information warfare manifests itself in the light of these two internal / external national security crises. Three hypotheses that guide this research report are: Russian pattern of information warfare has a long tradition which can be traced back to the Cold War era; it is possible to discern specifically Russian, partially divergent information warfare pattern; and finally by exploring the two recent internal / external national security crises, it becomes possible to sketch specifically Russian systematics. In this research report the main focus of interest is on the information-psychological dimension of the overall information warfare concept as part of the military science tradition. After such theoretical review the two empirical cases will be contextualized and chronologically introduced. Analysis will be sharpened on the parties’ actions especially from the information-psychological perspective. This will be done with the help of the developed Russia’s six action fields-model which has been divided into two main dimensions: political and military with three levels: strategic informa-tion-psychological level, and two tactical levels, namely information-technical and information-PSYOP. This creates six possible actions fields. As the empirical analysis will reveal, many of these six action fields have been used by Russia in its internal / external national security crises, which proves the study’s hypotheses: Russia has its own pattern of information psychologic warfare that is based on its historical tradition and as such it creates a base for Russian systematics.

Taxonomy, species richness and biogeography of Finnish crane flies (Diptera, Tipuloidea).

Biodiversity is unequally spread throughout terrestrial ecosystems. The highest species richness of animals and plants is encountered around the Equator, and naturalists observe a decrease in the number of creatures with increasing latitude. Some animal groups, however, display an anomalous species richness pattern, but these are exceptions to the general rule. Crane flies (Diptera, Tipuloidea) are small to large sized, non-biting nematoceran insects, being mainly associated with moist environments. The species richness of crane flies is highest in the tropics, but these insects are species rich and abundant in all biogeographic realms, boreal and arctic biomes included. The phylogeny and systematics of crane flies are still at an early stage and somewhat controversial. New species are constantly discovered even from temperate Europe, faunistically the best known continent. Crane flies have been rather neglected group of insects in Finland. The history of Finnish crane fly taxonomy and faunistics started in 1907, the year when Carl Lundström published his two first articles on tipuloids. Within roughly 100 years there have been only a handful of entomologists studying the Finnish fauna, and the species richness and natural history of these flies have remained poorly understood and mapped. The aim of this thesis is to clarify the taxonomy of Finnish crane flies, present an updated and annotated list of species and seek patterns in regional species richness and assemblage composition. Tipula stackelbergi Alexander has been revised (I). This species was elevated to a species rank from a subspecific rank under T. pruinosa Wiedemann and T. stackelbergi was also deleted from the list of European crane flies. Two new synonyms were found: T. subpruinosa Mannheims is a junior synonym of T. freyana Lackschewitz and T. usuriensis Alexander is a junior synonym of T. pruinosa. A new species Tipula recondita Pilipenko & Salmela has been described (II). Both morphology and COI (mtDNA) sequences were used in the assessment of the status of the species. The new species is highly disjunct, known from Finland and Russian Far East. A list of Finnish crane flies was presented, including the presence of species in the Finnish biogeographical provinces (III). A total of twenty-four species were formally reported for the first time from Finland and twenty-two previously reported species were deleted from the list. A short historical review on the studies of Finnish crane flies has been provided. The current list of Finnish species consists of 338 crane flies (IV, Appendix I). Species richness of all species and saproxylic/fungivorous species is negatively correlated with latitude, but mire-dwelling species show a reversed species richness gradient (i.e. an increase in the number of species toward north). Provincial assemblages displayed a strong latitudinal gradient and faunistic distance increased with increasing geographical distance apart of the provinces. Nearly half (48 %) of the Finnish crane flies are Trans-Palaearctic, roughly one-third (34 %) are West Palaearctic and only 16 and 2 % are Holarctic and Fennoscandian, respectively. Due to the legacy of Pleistocene glaciations, endemic Fennoscandian species are problematic and it is thus concluded that there are probably no true endemic crane flies in this region. Finally, there are probably species living within Finnish borders that have hitherto remained unnoticed. Based on subjective assessment, the number of “true” (i.e. recorded + unknown species) species count of Finnish crane flies is at minimum 350.

Development and application of polymerase chain reaction test for detection of Conidiobolus lamprauges

Conidiobolomycosis is a granulomatous disease caused by the fungus Conidiobolus spp. in humans and animals. Traditional technique for diagnosis of the disease is isolation of the agent associated with the presence of typical clinical signs and pathological conditions. The aim of this study was to describe the development of a specific polymerase chain reaction (PCR) test for Conidiobolus lamprauges to detect the fungus in clinical samples. Samples from suspected animals were collected and submitted to isolation, histopathological analysis and amplification by PCR. DNA from tissues was subjected to PCR with fungi universal primers 18S rDNA gene, and specific primers were designed based on the same gene in C. lamprauges that generated products of about 540 bp and 222 bp respectively. The culture was positive in 26.6% of clinical samples. The PCR technique for C. lamprauges showed amplification of DNA from fresh tissues (80%) and paraffin sections (44.4%). In conclusion, the PCR technique described here demonstrated a high sensitivity and specificity for detection of fungal DNA in tissue samples, providing a tool for the rapid diagnosis of C. lamprauges.

Kolme kaupallista analyysiohjelmaa ekologeille

Tietokoneohjelman arvostelu.

Tietojohtaminen projektissa: hankevastaavan ja projektipäällikön näkökulmia

Tutkimuksen tavoitteena oli selvittää miten tietojohtamisen eri osatekijät näkyvät projektin läpiviemisen aikana. Teoreettinen viitekehys kokoaa projektijohtamisen osatekijät ja tietojohtamisen kolme pääelementtiä (tiedon hankinta, tiedon säilyttäminen ja tiedon siirto) yhteen projektin elinkaaren ympärille. Tutkimus toteutettiin laadullisena tutkimuksena, jonka empiirinen osuus suoritettiin haastattelemalla projektin asiakkaan vastuuhenkilöitä sekä toimittajan vastuuhenkilöitä yksi kerrallaan. Näiden lisäksi suoritettiin ryhmähaastattelu kohderyhmän portfoliohallitsijoiden kesken. Empiriasta saadut tulokset mallinnettiin viitekehyksen mukaisesti tietojohtamisen pääelementeille. Tiedon hankinnassa projektijohtamisen kentästä korostui sidosryhmät ja riskit, tiedon säilyttämisessä korostui integrointi ja viestintä sekä tiedon siirtämisessä korostui aika ja HR. Projektin aikana luodun ja hankitun osaamisen ja oppien säilyttäminen koettiin haastateltavien mukaan tietojohtamisen osatekijöistä haastavimmaksi. Alatutkimuskysymyksenä selvitetyt esteet ja hidasteet tietojohtamisen menetelmille kulminoituivat haastatteluissa prosesseihin, vastuisiin ja systemaattisuuden puuttumiseen. Haastatteluiden ja kirjallisuustutkimuksen tuloksena tehtiin kohdeyritykseen suosituksia tietojohtamisen elementtien hyödyntämisessä projekteissa.

Pankin notariaattipalvelut – asiantuntijan vastuu sivullista kohtaan

Pro gradu -tutkimuksen keskeisin tavoite on selvittää perusteita pankin notariaattipalveluiden tarjoajan vastuun laajenemiselle yli välittömän toimeksiantosopimussuhteen. Tutkimus on rajattu vain Suomen oikeuteen ja tutkimusasetelmasta seuraa, että vastuun henkilöllisen ulottuvuuden laajentumisen tilanteista tutkimuksessa perehdytään nimenomaisesti sivullisintressitilanteisiin. Tutkimus on toteutettu lainopillisesti. Suomen laissa ei säädetä suoraan välittömän sopimussuhteen ylittävästä vastuusta, joten lainsäädännön jäädessä vähemmälle korostuvat tutkimuksessa sopimusoikeuden yleiset periaatteet sekä korkeimman oikeuden tulkintakäytäntö ja oikeustieteellinen tutkimus. Tämä johtaa etsimään ja jäsentämään tutkimusongelman kannalta merkityksellisiä oikeusperiaatteita ja reaalisia argumentteja tarkkojen tulkintasuositusten tarjoamisen sijaan. Tutkimus osoittaa, että välittömän sopimussuhteen ylittävää vastuuta tulee arvioida sopimusvastuun normien mukaisesti deliktivastuun sijaan, ja sitä voidaan pitää eräänä sopimusvastuun erityistapauksena. Tutkimuksen pohjalta voidaan todeta, että pankin notariaattipalveluiden tarjoajan vastuu voi ulottua yli toimeksiantosopimussuhteen, jos vastuun syntymistä voidaan puoltaa riittävillä reaalisilla argumenteilla. Tällaisia perusteita ovat muun muassa pankin notariaattipalvelun tarjoajan ja sivullisen välinen erityissuhde ja pankin notariaattipalvelun tarjoajan mahdollisuus ennakoida vahinkoriski. Lisäksi kaksiasianosaissuhteen rajoituksesta poikkeamista voidaan perustella vaihdannan reaalisen rakenteen huomioonottamisella sekä sivulliselle syntyneellä perustellun luottamuksen suojaamisella.

The nucleolus: a paradigm for cell proliferation and aging

The nucleolus is the cellular site of ribosome biosynthesis. At this site, active ribosomal DNA (rDNA) genes are rapidly transcribed by RNA polymerase I (pol I) molecules. Recent advances in our understanding of the pol I transcription system have indicated that regulation of ribosomal RNA (rRNA) synthesis is a critical factor in cell growth. Importantly, the same signaling networks that control cell growth and proliferation and are deregulated in cancer appear to control pol I transcription. Therefore, the study of the biochemical basis for growth regulation of pol I transcription can provide basic information about the nuclear signaling network. Hopefully, this information may facilitate the search for drugs that can inhibit the growth of tumor cells by blocking pol I activation. In addition to its function in ribosome biogenesis, recent studies have revealed the prominent role of the nucleolus in cell senescence. These findings have stimulated a new wave of research on the functional relationship between the nucleolus and aging. The aim of this review is to provide an overview of some current topics in the area of nucleolus biology, and it has been written for a general readership.

Partial characterization of nif genes from the bacterium Azospirillum amazonense

Azospirillum amazonense revealed genomic organization patterns of the nitrogen fixation genes similar to those of the distantly related species A. brasilense. Our work suggests that A. brasilense nifHDK, nifENX, fixABC operons and nifA and glnB genes may be structurally homologous to the counterpart genes of A. amazonense. This is the first analysis revealing homology between A. brasilense nif genes and the A. amazonense genome. Sequence analysis of PCR amplification products revealed similarities between the amino acid sequences of the highly conserved nifD and glnB genes of A. amazonense and related genes of A. brasilense and other bacteria. However, the A. amazonense non-coding regions (the upstream activator sequence region and the region between the nifH and nifD genes) differed from related regions of A. brasilense even in nitrogenase structural genes which are highly conserved among diazotrophic bacteria. The feasibility of the 16S ribosomal RNA gene-based PCR system for specific detection of A. amazonense was shown. Our results indicate that the PCR primers for 16S rDNA defined in this article are highly specific to A. amazonense and can distinguish this species from A. brasilense.

Low variation in ribosomal DNA and internal transcribed spacers of the symbiotic fungi of leaf-cutting ants (Attini: Formicidae)

Leaf-cutting ants of the genera Atta and Acromyrmex (tribe Attini) are symbiotic with basidiomycete fungi of the genus Leucoagaricus (tribe Leucocoprineae), which they cultivate on vegetable matter inside their nests. We determined the variation of the 28S, 18S, and 5.8S ribosomal DNA (rDNA) gene loci and the rapidly evolving internal transcribed spacers 1 and 2 (ITS1 and ITS2) of 15 sympatric and allopatric fungi associated with colonies of 11 species of leafcutter ants living up to 2,600 km apart in Brazil. We found that the fungal rDNA and ITS sequences from different species of ants were identical (or nearly identical) to each other, whereas 10 GenBank Leucoagaricus species showed higher ITS variation. Our findings suggest that Atta and Acromyrmex leafcutters living in geographic sites that are very distant from each other cultivate a single fungal species made up of closely related lineages of Leucoagaricus gongylophorus. We discuss the strikingly high similarity in the ITS1 and ITS2 regions of the Atta and Acromyrmex symbiotic L. gongylophorus studied by us, in contrast to the lower similarity displayed by their non-symbiotic counterparts. We suggest that the similarity of our L. gongylophorus isolates is an indication of the recent association of the fungus with these ants, and propose that both the intense lateral transmission of fungal material within leafcutter nests and the selection of more adapted fungal strains are involved in the homogenization of the symbiotic fungal stock.

Green ICT metrics and Biomimicry

Sustainability issue of ICT have gathered attention in recent years, and researchers are working on this problem. Sustainability incorporates numerous interconnected aspects as well as methods to achieve it in ICT, so it is quite complicated to have a general view on a problem without a proper framework. However, a general methodology for such a research is missing. In this work it is proposed to use Biomimicry approach as a framework for sustainability research and development, as it introduces systematics and also forces to account sustainable aspects. Additionally, an interesting problem of green network measurements for enhancing sustainability in ICT will be researched using mentioned approach. The goal is to investigate Biomimicry as a systemic approach for developing sustainable systems, as well as to apply it in green network measurements study. Comparative study is performed for examining Biomimicry approach, as well as a use case of green network measurements research is presented. As a result, green network measurement can potentially improve sustainability, but only to a limited extent as it cannot incorporate all the aspects; within Biomimicry approach, two methodologies exist. It is possible to conclude that Biomimicry is a good framework for developing sustainable systems, nevertheless, another methodology has to be introduced within it; new methodology has to incorporate advantages of two existing ones.

Screening of iron-enriched fungus from natural environment and evaluation of organically bound iron bioavailability in rats

Iron is an essential element for nearly all living organisms, and its deficiency is the most common form of malnutrition in the world. The organic forms of trace elements are considered more bioavailable than the inorganic forms. Although Saccharomyces cerevisiae can enrich metal elements and convert inorganic iron to organic species, its tolerability and transforming capacity are limited. The aim of this study was to screen higher biomass and other iron-enriched fungi strains besides Saccharomyces cerevisiae from the natural environment. A PDA medium containing 800 μg/mL iron was used for initial screening. Fifty strains that tolerated high iron concentration were isolated from the natural environment, and only one strain, No.BY1109, grew well at Fe (II) concentration of 10,000μg/ml. According to morphological characterization, 18S rDNA sequence analysis, and biophysical and biochemical characterization, the strain No.BY1109 was identified as Rhodotorula. The iron content of No.BY1109 (10 mg Fe/g dry cell) was determined using atomic absorption spectrometry. The results of distribution of iron in the cells showed that iron ion was mainly chelated in the cell walls and vacuoles. The bioavailability in rats confirmed that strain No.BY1109 had higher absorption efficiency than that of ferrous sulfate after single dose oral administration. The present study introduces new iron supplements, and it is a basis for finding new iron supplements from natural environment.

Amplification of the DFR1 gene in Saccaromyces cerevisiae

A new system was employed to study amplification of t,he DHF'R gene DFB,1 ) in Sa<,:;charoillYCB§. .Q~~Yi...S!i<;1~. . This system consists of a series of yeast strains containing a casset,te which encodes t he yeast, D..ERl gene ttghtly linked tjO a f usion of the yeast 1EU2. regulat,ory region wi tJ1 the LAQZ str ctural gene from E. cO.1-1 (,) . M. Clement , unpubl i,::;hed) . Th's casset;t e was shown t.o be integrat,ed int o a unj que chromosomal l ocati on in each strain . Yeast cells were se l ected for MTX-resistance and overproduction of ~ galac t osi d se ( B-gal ). Since the inserted DF'Rl and ~ACZ genes are independently regulated, it was thought that cel l s with this phenotype probably contain e d ampl if ications of the cassette. A lar ge variat ion in the f requn y o f MTX-resistance was found between the di ff e r ent str ains. These freqlen c ~ es r anged from about 2 x 10 - 7 fo r a population of cells containing the cassette integrated at, the BI J2.l gene in t,he middle of the long arm of chromosome V, to about 5 x 10-4 for a strain with the cassette i nserted in the r DNA cluster Abo It 85% of the MTX- res i stcmt iso l ates examined showed enhanced B·-gal act i v ity rel a t ive t o the parental strain . For the ma jorit y of strains, the mean B- gal activity in drug-r sistant clones was about 3 times that o f the parent following a single se l ect i on step . I n con t r ast, primary MTX-resistant derivat~ves of cells with the cassette inserted 3 at the rDNA cluster showed inc r eases in B- gal activity ranging from 9 - 14 f old r elative to the parent. Analysis of the latte r s train by Southe rn hybr idization indicated that the cassette was inde e d amplified several fold in MTX-re sistant derivatives. A sing l e strain, in which the cassette was inserted at the !lEA;], loc u.s , was used to examine in more detai 1 , the parameters affecting DFRl gene amplificat~ion in yeast . The mean B- gal activity in drug-resistant derivatives of this strain could be increased from 3 to 6 or 7 fold relative to the parent, by stepwise sel ection using increasing MTX concentrations. B-gal overproduction was found to be un stable in all primary and highly -resistant isolates examined. There was no indication, h owever, of a decrease i n growth r a t e in MTX-res i s tant cells which overproduced B - gal.

Isolation and analysis of a corprinus cinereus DNA fragment showing homology to a fimbrial cDNA from ustilago violacea

Surface proteinaceous fibrils, termed fimbriae, were first identified on gram negative bacteria in the 1940s. Fungal fimbriae, discovered some 25 years later, are found on members of all fungal classes. In the present study, polyclonal antiserum raised against the fimbrial proteins of U. vio/acea were used in order to identify antigenically related proteins from Coprinus cinereus and Schizophy//um commune. Two polypeptides with molecular masses of 37 and 39 kDa from C. cinereus were observed and confirm earlier results. A single previously unidentified 50 kDa polypeptide in S. commune crossreacted with the antiserum. The 50 kDa protein was found to consist of 3 isoforms with isoelectric points ranging from 5.6 to 5.8. A fimbrial cDNA derived from U. vio/acea was used to identify DNA restriction fragments from C. cinereus and S. commune showing homology to the fimbrial transcript of U. vio/acea. Heterologous hybridization with this cDNA was used in order to screen a C. cinereus genomic DNA library. A single clone, A2-3A, with a 14 kbp insert showed strong homology to the pfim3-1 cDNA. The region of homology, a 700 bp Xba I fragment, was subcloned into pUG19. This plasmid was refered to as pXX8. DNA sequence determinations of pXX8 and adjacent fragments from A2-3A suggested that the cloned DNA was a portion of the rONA repeat encoding the small subunit rRNA. DNA sequence analysis of pfim3-1 yielded an incomplete open reading frame. The predicted amino acid sequence codes for a 206 amino acid, 22 kDa polypeptide which contains a domain similar to a transmembrane domain from rat leukocyte antigen, GDS3. As well, an untranslated 576 nucleotide domain showed 81 % homology to pXX8 and 830/0 homology to the 188 rRNA sequence of Ustilago maydis. This sequence was found adjacent to a region of adenine-thymine base pairs presumed to represent the polyadenylation sequence of the fimbrial transcript. The size and extent of homology is sufficient to account for the hybridization of pfim3-1 to rDNA. It is suggested that this domain represents a completely novel regulatory domain within eukaryotes that may enable the observed rapid regeneration of fimbriae in U. violacea.

Study of new yeast strains as novel starter cultures for Riesling icewine production

Icewine is a sweet dessert wine fermented from the juice of grapes naturally frozen on the vine. The production of Icewine faces many challenges such as sluggish fermentation, which often yields wines with low ethanol, and an accumulation of high concentration of volatile acidity, mainly in the form of acetic acid. This project investigated three new yeast strains as novel starter cultures for Icewine fermentation with particular emphasis on reducing acetic acid production: a naturally occurring strain of S. bayanus/S. pastorianus isolated from Icewine grapes, and two hybrids between S. cerevisiae and S. bayanus, AWRI 1571 and AWRI 1572. These strains were evaluated for sugar consumption patterns and metabolic production of ethanol, glycerol and acetic acid, and were compared to the performance of a standard commercial wine yeast KI-VI116. The ITS rONA region of the two A WRI crosses was also analyzed during fermentations to assess their genomic stability. Icewine fermentations were performed in sterile filtered juice, in the absence of indigenous microflora, and also in unfiltered juice in order to mirror commercial wine making practices. The hybrid A WRI 1572 was found to be a promising candidate as a novel starter culture for Icewine production. I t produced 10.3 % v/v of ethanol in sterile Riesling Icewine fermentations and 11.2 % v/v in the unfiltered ones within a reasonable fermentation time (39 days). Its acetic acid production per gram sugar consumed was approximately 30% lower in comparison with commercial wine yeast K I -V 1116 under both sterile filtered and unfiltered fermentations. The natural isolate S. bayanus/S. pastorianus and AWRI 1571 did not appear to be suitable for commercial Icewine production. They reached the target ethanol concentration of approximately 10 % v/v in 39 day fermentations and also produced less acetic acid as a function of both time and sugar consumed in sterile fermentations compared to KI-V1116. However, in unfiltered fermentations, both of them failed to produce the target concentration of ethanol and accumulated high concentration of acetic acid. Both A WRI crosses displayed higher loss of or reduced copies in ITS rDNA region from the S. bayanus parent compared to the S. cerevisiae parent; however, these genomic losses could not be related to the metabolic profile.