The viral coat protein is regulated by HSP70 and HSP40 in Potato virus A infection

Plus-stranded (plus) RNA viruses multiply within a cellular environment as tightly integrated units and rely on the genetic information carried within their genomes for multiplication and, hence, persistence. The minimal genomes of plus RNA viruses are unable to encode the molecular machineries that are required for virus multiplication. This sets requisites for the virus, which must form compatible interactions with host components during multiplication to successfully utilize primary metabolites as building blocks or metabolic energy, and to divert the protein synthesis machinery for production of viral proteins. In fact, the emerging picture of a virus-infected cell displays tight integration with the virus, from simple host and virus protein interactions through to major changes in the physiological state of the host cell. This study set out to develop a method for the identification of host components, mainly host proteins, that interact with proteins of Potato virus A (PVA; Potyvirus) during infection. This goal was approached by developing affinity-tag based methods for the purification of viral proteins complexed with associated host proteins from infected plants. Using this method, host membrane-associated viral ribonucleoprotein (RNP) complexes were obtained, and several host and viral proteins could be identified as components of these complexes. One of the host proteins identified using this strategy was a member of the heat shock protein 70 (HSP70) family, and this protein was chosen for further analysis. To enable the analysis of viral gene expression, a second method was developed based on Agrobacterium-mediated virus genome delivery into plant cells, and detection of virally expressed Renilla luciferase (RLUC) as a quantitative measure of viral gene expression. Using this method, it was observed that down-regulation of HSP70 caused a PVA coat protein (CP)-mediated defect associated with replication. Further experimentation suggested that CP can inhibit viral gene expression and that a distinct translational activity coupled to replication, referred to as replication-associated translation (RAT), exists. Unlike translation of replication-deficient viral RNA, RAT was dependent on HSP70 and its co-chaperone CPIP. HSP70 and CPIP together regulated CP turnover by promoting its modification by ubiquitin. Based on these results, an HSP70 and CPIP-driven mechanism that functions to regulate CP during viral RNA replication and/or translation is proposed, possibly to prevent premature particle assembly caused by CP association with viral RNA.

Streptomyces scabies and S. turgidiscabies as pathogens causing potato common scab, and potential methods for their control

Tieteellinen tiivistelmä Common scab is one of the most important soil-borne diseases of potato (Solanum tuberosum L.) in many potato production areas. It is caused by a number of Streptomyces species, in Finland the causal agents are Streptomyces scabies (Thaxter) Lambert & Loria and S. turgidiscabies Takeuchi. The scab-causing Streptomyces spp. are well-adapted, successful plant pathogens that survive in soil also as saprophytes. Control of these pathogens has proved to be difficult. Most of the methods used to manage potato common scab are aimed at controlling S. scabies, the most common of the scab-causing pathogens. The studies in this thesis investigated S. scabies and S. turgidiscabies as causal organisms of common scab and explored new approaches for control of common scab that would be effective against both species. S. scabies and S. turgidiscabies are known to co-occur in the same fields and in the same tuber lesions in Finland. The present study showed that both these pathogens cause similar symptoms on potato tubers, and the types of symptoms varied depending on cultivar rather than the pathogen species. Pathogenic strains of S. turgidiscabies were antagonistic to S. scabies in vitro indicating that these two species may be competing for the same ecological niche. In addition, strains of S. turgidiscabies were highly virulent in potato and they tolerated lower pH than those of S. scabies. Taken together these results suggest that S. turgidiscabies has become a major problem in potato production in Finland. The bacterial phytotoxins, thaxtomins, are produced by the scab-causing Streptomyces spp. and are essential for the induction of scab symptoms. In this study, thaxtomins were produced in vitro and four thaxtomin compounds isolated and characterized. All four thaxtomins induced similar symptoms of reduced root and shoot growth, root swelling or necrosis on micro-propagated potato seedlings. The main phytotoxin, thaxtomin A, was used as a selective agent in a bioassay in vitro to screen F1 potato progeny from a single cross. Tolerance to thaxtomin A in vitro and scab resistance in the field were correlated indicating that the in vitro bioassay could be used in the early stages of a resistance breeding program to discard scab-susceptible genotypes and elevate the overall levels of common scab resistance in potato breeding populations. The potential for biological control of S. scabies and S. turgidiscabies using a non-pathogenic Streptomyces strain (346) isolated from a scab lesion and S. griseoviridis strain (K61) from a commercially available biocontrol product was studied. Both strains showed antagonistic activity against S. scabies and S. turgidiscabies in vitro and suppressed the development of common scab disease caused by S. turgidiscabies in the glasshouse. Furthermore, strain 346 reduced the incidence of S. turgidiscabies in scab lesions on potato tubers in the field. These results demonstrated for the first time the potential for biological control of S. turgidiscabies in the glasshouse and under field conditions and may be applied to enhance control of common scab in the future.

Resistance responses blocking systemic movement of Potato virus A in potato

Peruna kestää A-virusta estämällä sen leviämistä Peruna on maissin ohella maailman kolmanneksi tärkein ravintokasvi vehnän ja riisin jälkeen. Perunaa lisätään kasvullisesti mukuloita istuttamalla, jolloin virukset siirtyvät sairaiden siemenmukuloiden välityksellä kasvukaudesta toiseen. Virustauteja voi torjua ainoastaan terveen siemenperunan ja kestävien lajikkeiden avulla. Kestävyys perustuu usein siihen, että kasvi estää viruksen leviämisen tartuntakohdasta välttyäkseen virustaudilta. Tässä työssä tutkittiin kolmea perunan A-viruksen (PVA) liikkumista estävää kestävyysmekanismia perunassa. Lisäksi työn kokeelliseen osaan oleellisesti kuuluvaa virustartutusta varten kehitettiin uusi paranneltu versio geenipyssystä. Tämä itse rakennettu laite optimoitiin PVA:n tartuttamiseen mahdollisimman helposti ja pienin käyttökustannuksin. Tutkimuksen kohteena olleessa perunan risteytysjälkeläistössä oli PVA:ta kestäviä kasveja (ryhmä nnr), jotka estivät viruksen liikkumisen aiheuttamatta oireita tartutuskohdassa, sekä kasveja, joissa PVA aiheutti kuolioläikkinä näkyvän yliherkkyysvasteen (ryhmä HR). Molemmissa kestävyystyypeissä virus pystyi monistumaan ja leviämään solusta soluun paikallisesti, mutta liikkuminen muihin kasvinosiin nilan kautta estyi. Ryhmän nnr kasveissa PVA-tartunta ei aiheuttanut tilastollisesti merkitsevää muutosta useimpien geenien ilmenemiseen tartuntakohdassa. Ainoastaan geeniperhe, joka ilmentää tiettyä proteinaasi-inhibiittoria (PI), reagoi PVA:han 24 tuntia tartutuksesta. Kun tämän PVA:han reagoivan geeniperheen jäsenet hiljennettiin nnr- perunalinjoissa, ne muuttuivat alttiiksi PVA:lle ja virus levisi tartuntakohdasta muihin kasvinosiin. Tulos osoittaa, että PI on viruskestävyystekijä. Lisäksi muut tutkimuksessa saadut tulokset tukevat mahdollisuutta, että PI estää PVA:n P1-proteinaasin toimintaa. HR-linjoissa todettiin erilaisiin puolustusvasteisiin liittyvien PR-geenien aktivoitumista PVA-tartunnan seurauksena, mutta myös ilman sitä kasvien kasvettua mullassa noin neljä viikkoa. Sen sijaan solukkoviljelyssä tai vasta kaksi viikkoa mullassa kasvaneissa kasveissa vastaavaa ei vielä todettu. Tulos viittaa siihen, että HR-perunat reagoivat herkemmin ympäristöön ja/tai kasvin kehitysasteeseen laukaisten puolustusvasteita, jotka saattavat parantaa kestävyyttä taudinaiheuttajia vastaan. Kolmas tutkittu kestävyystyyppi havaittiin Pito-perunalajikkeessa. Se muistutti nnr-kestävyyttä siten, että myös siinä viruksen liikkuminen nilassa muihin kasvinosiin estyi. PVA:n todettiin pysähtyvän vasta lehtiruodin tyvelle muodostuvaan irtoamisvyöhykkeeseen, mitä havainnollistettiin käyttämällä muunnettua PVA-rotua, joka tuotti UV-valossa fluoresoivaa vihreää valoa. Tulos viittaa siihen, että virus ei pääse kulkemaan vyöhykkeeseen kuuluvan suojaavan kerroksen läpi, jollei sillä ole pääsyä nilaan. Tällainen kestävyys on tarpeen, jotta virus ei voi korvata nilakuljetusta solusta soluun leviämisellä. Tulokset tuovat uusia näkökulmia kasvien viruskestävyyteen ja auttavat selittämään viruksen nilakuljetuksen estymistä sekä solusta soluun leviämisen pysähtymistä kestävissä kasveissa.

Adoption processes for EDM, EDI and BIM technologies in the construction industry

Three strategically important uses of IT in the construction industry are the storage and management of project documents on webservers (EDM), the electronic handling of orders and invoices between companies (EDI) and the use of 3-D models including non-geometrical attributes for integrated design and construction (BIM). In a broad longitudinal survey study of IT use in the Swedish Construction Industry the extent of use of these techniques was measured in 1998, 2000 and 2007. The results showed that EDM and EDI are currently already well-established techniques whereas BIM, although it promises the biggest potential benefits to the industry, only seems to be at the beginning of adoption. In a follow-up to the quantitative studies, the factors affecting the decisions to implement EDM, EDI and BIM as well as the actual adoption processes, were studied using semi-structured interviews with practitioners. The theoretical basis for the interview studies was informed by theoretical frameworks from IT-adoption theory, where in particular the UTAUT model has provided the main basis for the analyses presented here. The results showed that the decisions to take the above technologies into use are made on three differ- ent levels: the individual level, the organizational level in the form of a company, and the organiza- tional level in the form of a project. The different patterns in adoption can to some part be explained by where the decisions are mainly taken. EDM is driven from the organisation/project level, EDI mainly from the organisation/company level, and BIM is driven by individuals pioneering the technique.

Building Information Modelling – Experts´ Views on Standardisation and Industry Deployment

The goal of the single building information model has existed for at least thirty years and various standards have been published leading up to the ten-year development of the Industry Foundation Classes. These have been initiatives from researchers, software developers and standards committees. Now large property owners are becoming aware of the benefits of moving IT tools from specific applications towards more comprehensive solutions. This study addresses the state of Building Information Models and the conditions necessary for them to become more widely used. It is a qualitative study based on information from a number of international experts and has asked a series of questions about the feasibility of BIMs, the conditions necessary for their success, and the role of standards with particular reference to the IFCs. Some key statements were distilled from the diverse answers received and indicate that BIM solutions appear too complex for many and may need to be applied in limited areas initially. Standards are generally supported but not applied rigorously and a range of these are relevant to BIM. Benefits will depend upon the building procurement methods used and there should be special roles within the project team to manage information. Case studies are starting to appear and these could be used for publicity. The IFCs are rather oversold and their complexities should be hidden within simple-to-use software. Inevitably major questions remain and property owners may be the key to answering some of these. A framework for presenting standards, backed up by case studies of successful projects, is the solution proposed to provide better information on where particular BIM standards and solutions should be applied in building projects.

Use of Document Management Systems - a Case Study of the Finnish Construction Industry

Triggered by the very quick proliferation of Internet connectivity, electronic document management (EDM) systems are now rapidly being adopted for managing the documentation that is produced and exchanged in construction projects. Nevertheless there are still substantial barriers to the efficient use of such systems, mainly of a psychological nature and related to insufficient training. This paper presents the results of empirical studies carried out during 2002 concerning the current usage of EDM systems in the Finnish construction industry. The studies employed three different methods in order to provide a multifaceted view of the problem area, both on the industry and individual project level. In order to provide an accurate measurement of overall usage volume in the industry as a whole telephone interviews with key personnel from 100 randomly chosen construction projects were conducted. The interviews showed that while around 1/3 of big projects already have adopted the use of EDM, very few small projects have adopted this technology. The barriers to introduction were investigated through interviews with representatives for half a dozen of providers of systems and ASP-services. These interviews shed a lot of light on the dynamics of the market for this type of services and illustrated the diversity of business strategies adopted by vendors. In the final study log files from a project which had used an EDM system were analysed in order to determine usage patterns. The results illustrated that use is yet incomplete in coverage and that only a part of the individuals involved in the project used the system efficiently, either as information producers or consumers. The study also provided feedback on the usefulness of the log files.

The Context and Diffusion of Knowledge in the Finnish Jewellery Industry - The role of the House of Fabergé

Researchers within the fields of economic geography and organizational management have extensively studied learning and the prerequisites and impediments for knowledge transfer. This paper combines two discourses within the two subjects: the-communities-of-practice and the learning region approaches, merging them through the so-called ecology of knowledge-approach, which is used to examine the knowledge transfer from the House of Fabergé to the Finnish jewellery industry. We examine the pre-revolution St Petersburg jewellery cluster and the post-revolution Helsinki, and the transfer of knowledge between these two locations through the components of communities of people, institutions and industry. The paper shows that the industrial dynamics of the Finnish modern-day goldsmith industry was inherently shaped both through the transfer and the non-transfer of knowledge. It also contends that the “knowledge-economy” is not anchored in and exclusive for the high technology sector of the late 20th century.

Dimensionality of quality from a customer perspective in the wood industry

The research analyzes product quality from a customer perspective in the case of the wood products industry. Of specific interest is to understand better how environmental quality is perceived from a customer perspective. The empirical material used comprises four data-sets from Finland, Germany and the UK, collected during 1992 2004. The methods consist of a set of quantitative statistical analyses. The results indicate that perceived quality from a customer perspective can be presented using a multidimensional and hierarchical construct with tangible and intangible dimensions, that is common to different markets and products. This applies in the case of wood products but also more generally at least for some other construction materials. For wood products, tangible product quality has two main sub-dimensions: technical quality and appearance. For product intangibles, a few main quality dimensions seem be detectable: Quality of intangibles related to the physical product, such as environmental issues and product-related information, supplier-related characteristics, and service and sales personnel behavior. Environmental quality and information are often perceived as being inter-related. Technical performance and appearance are the most important considerations for customers in the case of wood products. Organizational customers in particular also clearly consider certain intangible quality dimensions to be important, such as service and supplier reliability. The high technical quality may be considered as a license to operate , but product appearance and intangible quality provide potential for differentiation for attracting certain market segments. Intangible quality issues are those where Nordic suppliers underperform in comparison to their Central-European competitors on the important German markets. Environmental quality may not have been used to its full extent to attract customers. One possibility is to increase the availability of the environment-related information, or to develop environment-related product characteristics to also provide some individual benefits. Information technology provides clear potential to facilitate information-based quality improvements, which was clearly recognized by Finnish forest industry already in the early 1990s. The results indeed indicate that wood products markets are segmented with regard to quality demands