Strawberries are considered an important crop in California where in 2018 it was in the top 5 valued fruit and vegetable commodities valued at $2.84 billion accounting for 88% of the total U.S. production. Strawberry production can be severely impacted by soilborne pathogens that can affect strawberry roots, crowns and leaves which can result in plant mortality. As much as 50 to 60% mortality can occur in one field. Pathogens responsible for such losses include Colletotrichum acutatum (syn.C. nymphaeae), Macrophomina phaseolina and Verticillium dahliae. With the phaseout of methyl bromide, host resistance and an understanding of host-pathogen interactions can play an important role in control of these diseases. A two-year study was conducted in order to evaluate host resistance of anthracnose in 105 cultivars and elite breeding lines developed by six strawberry breeding programs. Cultivars and elite breeding lines were inoculated using three local isolates in both years. All breeding programs provided genotypes that had a wide range of anthracnose susceptibility ranging from 0 to 100% mortality during both years. In both years an average of 78% of all the plant mortality occurred by 1 January. From the 105 cultivars and elite breeding lines, 30 cultivars were common to both years. Of these 30 cultivars, nine of them differed in their disease susceptibility between experiments by more than 20%. This suggests that several years of field evaluation may be necessary to determine susceptibility to anthracnose. Popular cultivars that represent the spectrum of susceptibility are Monterey (susceptible), Festival (moderately resistant), and Sensation (resistant). A second study was conducted toevaluate pathogen colonization of resistant and susceptible strawberry cultivars, testing interactions among crown and root plant tissue and two sampling timings. These cultivars were challenged with two soilborne pathogens, Macrophomina phaseolinaand Verticillium dahliae,over two years. Existing qPCR protocols for M. phaseolina and V. dahliae were used in order to quantify how much pathogen DNA was detected in crown and root samples. For the 2016-2017 V. dahliae trial there were significant effects for cultivar. Cultivar Benicia had significantly higher pathogen DNA compared to resistant cultivars Marquis, UC-12 and Camino Real. Susceptible cultivar BG 1975 had significantly less pathogen DNA compared to resistant cultivars San Andreas and Petaluma. In the 2017-2018 V. dahliaetrial pathogen DNA amount was not significantly different based on cultivar, plant part colonization, or the sampling period. In the 2017-2018 M. phaseolina trial all three of the fixed factors, cultivars, plant part colonization and sampling period were statistically significant. Cultivar ‘Sweet Ann’ had a significantly higher level of M. phaseolinaDNA in the early vs. the late sampling.

The fungus Macrophomina phaseolina (Tassi) Goidanich (Mp) is the causal agent of charcoal rot in soybean and infects over 500 plant species worldwide. Mp produces various mycotoxins and is suspected of utilizing a toxin-mediated process to penetrate host tissue. Identification and evaluation of secondary metabolites produced by Mp will further elucidate the pathogenesis mechanisms used by the fungus. Mp cultures isolated from soybean were evaluated for phytotoxicity in a hydroponic soybean bioassay and chemically analyzed by LC-MS/MS. All Mp cultures at two dilutions induced phytotoxicity symptoms including chlorosis, necrosis, wilting, stunting, and death. Analysis identified 13 unreported secondary metabolites including mellein, a compound with various biological activities. The phytotoxicity of mellein was evaluated against soybean seedlings in hydroponic culture, and symptoms of wilting and stunting were observed at levels above 40 MUg/L. Observations suggest that mellein does not directly contribute to the phytotoxic effects of Mp cultures.

ROT-projekt allmänt är ganska återkommande i Sverige och utomlands. Till exempel spenderas drygt 38% av alla investeringar inom bostadsmarknaden på antingen renovering eller ombyggnation. Dessa projekt brukar ha många svårigheter och risker, bland annat oförutsägbarhet, bristfälliga handlingar/ritningar, många intressenter, hantering av kvarsittande hyresgäster samt grannar, brist på erfarenhet hos de olika parterna, mm… Tyvärr har dessa svårigheter varit en stor utmaning för byggentreprenörer under en lång period och väldigt få forskningar har försökt lösa dessa problem och optimera ombyggnationsprocessen. Det här examensarbetet kommer att försöka optimera ROT-projekt med hjälp av Lean och med fokus på standardisering. Författaren av detta examensarbete skulle nästan våga påstå att den här studien är väldigt unikt i sitt slag och kan räknas som ett pilotprojekt för tillämpning av standardisering inom ROT-projekt. För det här syftet har författaren samarbetat med en traditionell byggentreprenör där han fick vara med i ett ROT-projekt (fasadrenovering, byte av fönster, byte av värmestamm, mm…). Vissa arbetsmoment inom det ROT-projektet var repetitiva och då har kunnat metodstandarder tillämpas och resultatet av tillämpningen studerats och analyserats i denna rapport. Författaren har även intervjuat verksamma personer från byggentreprenörsidan samt beställarsidan och även en hyresgäst för att kunna jämföra resultaten från de äldre studierna om Lean och standardisering med hur tillämpas standardisering inom ROT-projekt. Resultatet från detta examensarbete visade att standardiseringen bidrar till att minska oförutsägbarheten inom ROT-projekt och hjälper byggentreprenören att ta fram en träffsäker tidplan samt budget. Det bidrar även till att uppnå den önskade kvalitén på det utförda arbetet samt minimera slöserierna inom byggbranschen som uppskattas till 30% av den totala produktionskostnaden. Resultaten visade också att det finns en stor brist på kunskap på hur kan Lean, särskilt standardisering tillämpas inom ROT-projekt. Resultaten har även visat att standardiseringen och det kontinuerliga förbättringsarbetet av standarderna inte leder till en bestående förbättring som fallet är när man sysslar med nyproduktion. Utan att de nya överraskningarna som drabbar produktionen leder till en viss försämring av resultatet, vilket gör att förbättringen inte blir bestående. Detta var säkert ett bevis på att det är dags att börja forska om tillämpningen av Lean och standardisering inom ROT-projekt, för de nuvarande studierna är baserade på nyproduktion och resultaten från dessa studier går inte att använda när man sysslar med ROT-projekt. Slutligen vill författaren gärna att forskarna börjar titta mer på det området och hoppas att det här examensarbetet blir som ögonöppnare för de. Författaren hoppas även att universitet och högskolor börjar undervisa separata ämnen som handlar om ROT-projekt och dess utmaningar och möjligheter. ROT-projekt är återkommande projekt som inte påverkas av konjunkturläget och branschen behöver säkert nyexaminerade ingenjörer som är kompetenta för att driva dessa projekt. / Renovation and remodeling projects in general are quite recurring in Sweden and abroad. For example, just over 38% of all investments in the housing market are spent on either renovation or remodeling. These projects usually have many difficulties and risks, including unpredictability, deficient documents / drawings, many stakeholders, managing of remaining tenants and neighbors, lack of experience on the part of the various parties, etc.… Unfortunately, these difficulties have been a major challenge for construction contractors for a long period and very little research has attempted to solve these problems and optimize the remodeling process. This thesis will try to optimize renovation and remodeling projects with the help of Lean and with a focus on standardization. The author of this thesis would almost dare to say that this study is unique in its kind and can be counted as a pilot project for the application of standardization in renovation and remodeling projects. For this purpose, the author has collaborated with a traditional building contractor where he was involved in a renovation and remodeling project (facade renovation, replacement of windows, replacement of heating installation, etc.…). Some work within the remodeling project were repetitive and then it was possible to apply method standards and the results of the application were studied and analyzed in this report. The author has also interviewed active people from the construction contractor side as well as the client side and a tenant to be able to compare the results from the older studies on Lean and standardization with how standardization is applied within renovation and remodeling projects. The results from this thesis showed that the standardization helps to reduce the unpredictability within renovation and remodeling projects and helps the construction contractor to develop an accurate schedule and budget. It also contributes to achieving the desired quality of the work performed and minimizing the waste in the construction industry, which is estimated at 30% of the total production cost. The results also showed that there is a great lack of knowledge on how Lean can be applied, especially standardization within renovation and remodeling projects. The results have also shown that the standardization and the continuous improvement of the standards do not lead to a permanent improvement, as is the case when dealing with new production. The new surprises that affect production leading to a certain deterioration of the result, which means that the improvement will not be consisting. This certainly proof that it is time to start researching the application of Lean and standardization in renovation and remodeling projects, because the current studies are based on new production and the results from these studies cannot be used when working with renovation and remodeling projects. Finally, the author would like the researchers to start looking more at that area and hope that this thesis will be an eye opener for them. The author also hopes that universities and colleges will start teaching separate subjects about renovation and remodeling projects, their challenges and opportunities. renovation and remodeling projects are recurring projects that are not affected by the economic situation and the industry certainly needs newly graduated engineers who are competent to run these projects.

Hydrogen is re-emerging as a serious alternative to fossil fuels. It is a clean gas with high energy density and its combustion only generates water vapour. Nevertheless, the hydrogen industry has a significant carbon footprint since this gas is mostly derived from fossil fuels reforming processes. This project focusses on the development of sustainable alternatives to conventional hydrogen production, in which approaches based on dark fermentation (DF) using an inexpensive residue from the brewery industry as primary feedstock are presented. Firstly, a fungal pre-treatment (FT) was proposed to degrade a high-strength brewery waste slurry (BWS) to obtain an effluent with a lower concentration of chemical oxygen demand (COD) but rich in readily fermentable sugars for the ensuing DF, thus improving hydrogen yields (HY). Secondly, microbial electrolysis and fuel cells (MECs and MFCs) were proposed to assist DF, generating electricity in MFCs while improving HY by MECs. Coupling both microbial electrochemical technologies sequentially after DF did not show any advantage. However, promising results were obtained for electricity and hydrogen production when taking a single-staged approach. Treating BWS directly by MFCs produced 2.0 watts/g COD consumed, while the DF process assisted simultaneously by MECs (DF/MEC) produced 1.6 times more hydrogen than DF alone. An average HY of 2.32 ± 0.06 mol H₂/mol glucose was attained between both DF/MEC and DF after FT, hence approaching the theoretical value of 2.4 mol H₂/mol glucose, representing roughly a 50% improvement compared to DF alone. With an overall COD reduction above 76%, the DF after FT exhibited the highest energy conversion rate per substrate consumed (6.3 kJ/g COD). As valuable by-products obtained, up to 31 g/L of fungal biomass, which is appreciated in many state-of-the-art biomaterials applications, was produced by using BWS. While in the DF/MEC process, 18 g/L of butyric acid were generated, which is three times more than with DF alone. Butyric acid being the precursor to butanol and building block of biodegradable thermoplastics, this result is not without significance. The proposed approaches not only valorize BWS but also validate their economic and environmental attractiveness as promising alternative hydrogen production methods.

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Diplodia ear rot (DER) of maize (Zea mays L.) caused by the fungus, Stenocarpella maydis (Berk.) Sutton has increased in incidence in localized fields over the past decade. My research focused on screening for resistance by examining the development of DER following inoculations prior to flowering, analyzing a diallel cross for DER resistance, and examining the genetic variability of the fungus from isolates collected from the U.S. and the Republic of South Africa. DER developed in maize following inoculations with a spore suspension prior to flowering in both greenhouse and field evaluations. A spore suspension gave a better differentiation of resistance responses than dried preparations of colonized millet, colonized ground popcorn, or kernels from a diseased maize ear, all applied in the whorl 10 to 15 days prior to flowering (V12 for inbreds), and natural occurrence of disease. General combining ability was significant for both 1994 and 1995 growing seasons in an analysis of the F₁ of the diallel cross, indicating that additive gene action may be responsible for resistance and could be introduced into commercial cultivars. Specific combining ability was significant in 1995 and indicates that dominant gene action or epistasis may play role in DER resistance. There were minimal numbers of isozyme polymorphisms found in my S. maydis collection. Two isolates were polymorphic for esterase, two isolates were polymorphic for hexokinase and malate dehydrogenase and one isolate was polymorphic for hexose kinase. Fungi that have limited isozyme polymorphisms often are biotrophs or fungi with formae speciales which are usually limited to one host. These groups of fungi usually have races and this may indicate that a gene-for-gene interaction exists. These findings suggest that i) the whorl inoculation separates genotypes into resistant, intermediate, and susceptible groupings; ii) additive gene action is predominant form of inheritance, and iii) there are few isozyme polymorphisms in the population of S. maydis sampled. / Ph. D.

Vitis vinifera L.(Vv), the European cultivated grapevine is one of the most worldwide important crops but is highly susceptible to the necrotrophic fungus Botrytis cinerea (Bc), the causal agent of bunch rot (BR) disease. In grapevine, as well as in other fruit species, it has been described a primary infection by Bc at full bloom, followed by a quiescence period during the berry development, and the fungal egression after veraison reaching the maximum at harvest. Today, this important disease is mainly controlled by massive use of fungicides, which are applied at different developmental stages that happen to be critical during the grapevine-Bc interaction. During the contact, the fungus must overcome several barriers from the host, which protect it from the pathogen attack and might be also modulated or activated due to the presence of the pathogen itself. The cuticle and the cell wall (CW) represent the first barriers from the plant encountered by the pathogen. To successfully colonize the plant tissue, Bc possesses several virulence factors, and CW modifying enzymes (CWMEs) are part of them. On the other hand, the regulated activity of the CWMEs, expressed both by the host and the pathogen, could alter the plant CW composition and porosity, therefore facilitating, or limiting the penetration of the fungus. Among the CWMEs, Pectin methylesterases (PMEs) regulate the degree of methyl esterification of the pectin homogalacturonan, also modifying the epitopes for the activity of other CWMEs such as polygalacturonases (PGs) and pectate lyases (PLs) by whose action, pectin becomes more susceptible to degradation and the CW more accessible by the pathogen. Previous works both in Arabidopsis thaliana and in crop species identified several PME genes with an altered expression in response to pathogens. A previous report characterizing Atpme17 mutant lines has highlighted a role of AtPME17 in the resistance response to Bc in contrast with the opposite role of AtPME3, suggesting that PME genes could have a completely different action during Bc response depending on the isoform involved. In this context, the main objective of the project was to identify new strategies for Bc BR-control, and specifically i) to identify candidate genes involved in the response to Bc, whose inactivation/overexpression would lead to Bc resistant plants and ii) to set up a molecular method to monitor the Bc load in the field and therefore implement a more sustainable control of the pathogen. To further understand the effects of the Bc primary infection in grapevine flowers at CW level, two contrasting genotypes (Souvignier gris (SG) and Teroldego (TE)) in their resistance to the fungus were considered. An artificial inoculation of different biological replicates, in vase maintained, was performed at full bloom, in controlled conditions, and samples were collected at 24 hours post-inoculation (hpi) with the fungus and post-treatment with the respective control, for the following RNA-seq analysis and biochemical characterization of PME activity and CW composition in the two genotypes before and upon infection. The Bc load was estimated in the flowers using qPCR and as expected, a higher biomass of Bc was found in TE, the susceptible cultivar, than in SG, the resistant one. The analysis of CW composition, PME activity and degree of pectin methyl-esterification, both in treated and control flowers, showed significant differences between the two genotypes, in particular SG showed a significant induction of PME activity with respect to the control, evidence not present in the susceptible genotype. The RNAseq analysis on the same samples showed a total of 4800 genes modulated, out of which 3064 are only modulated in TE, 739 only in SG and a common group of 997 genes. Regardless of the cultivar, upon infection there was a total 2919 genes upregulated vs 1909 genes downregulated. A gene set enrichment analysis (GSEA) indicated several over-represented categories upon infection, including response to pathogens and biosynthesis of secondary metabolites, with a general down-regulation of those genes related to CW organization and pectin modification (CWMEs), mostly in the resistant genotype. Within the down-regulated CWMEs, Pectin methylesterase (PME) genes were found highly represented. Unlike, a larger gene set, in many cases with a higher fold-change of induction, was identified in TE respect to SG. This is the case of genes involved in the defense response and its regulation, and in the modification/reinforcement of the cell wall, therefore attesting for an initial tentative by the susceptible genotype to counteract the pathogen, although at the end without success. This was also the case of the seven VviPME genes previously highlighted by the in-silico co-expression analysis and therefore of VviPME10, the gene with the highest homology to AtPME17. Among the regulators, one WRKY factor (VviWRKY3), known to be related to defense response in grapevine mediated by stilbene synthesis, was also further characterized as putative regulator of VviPME10, whose promoter hosts more than one several predicted binding sites for VviWRKY3. Indeed, luciferase assay results indicate a significant activation of VviPME10 promoter by VviWRKY3 factor. Parallelly, the genome-wide analysis of the last structural annotation of the grapevine genome assembly allowed us to identify 62 VviPME gene members, 15 more than a previous report, and manually curate the gene structure for 39 of them. Then, to corroborate the idea of the role of the CW, and in particular of PME activity, in the grapevine response to the fungus, an in silico co-expression analysis of the 62 VviPME members, considering the publicly available RNA-seq experiments related to grapevine-Bc interactions and the RNAseq experiment conducted in this project, was performed. The analysis highlighted a group of seven genes (VviPME1, VvPME9, VviPME10, VviPME11, VviPME12, VviPME13 and VviPME54) with significant induction upon Bc infection, five of them (VviPME8, VviPME9, VviPME10, VviPME11, and VviPME54) located in the same chromosome (chr06). VviPME10 showed the highest homology and was found to be phylogenetically close to the Arabidopsis thaliana PME17 gene, suggesting being considered as its putative orthologue. Afterward, Therefore, considering the increased VviPME10 expression upon infection, and the reported effect of AtPME17 in A. thaliana, VviPME10 was selected as a potential candidate to study its role in grapevine. In this regard, two strategies were adopted, i. VviPME10 knock-out (KO) with CRISPR/Cas9 and ii. VviPME10 overexpression (OE, under CaMV35S promoter) through embryogenic callus transformation of the grapevine cultivar ‘Sugraone’ mediated by Agrobacterium tumefaciens. More than 100 embryos developed, and around 20 plantlets per transformation were analyzed to check the presence of the transgenic construct. Then, the mutation profile, in the case of KO lines, and the expression analysis of the transgene, in the case of OE lines, were carried out to select the appropriate lines to acclimatize. OE lines were also tested for VviPME10 activity. A total protein extract was obtained from the leaves of the lines, showing a higher protein activity compared to the control, and indicating the functionality of the enzyme. Unfortunately, grapevine OE lines couldn’t be analyzed for their response to Bc, while KO lines showed a significantly larger lesion area when compared to the control at 5 days post fungal inoculation (dpi). However, the effect of VviPME10 overexpression upon Bc infection was evaluated also in Nicotiana benthamiana VviPME10-OE lines, generated in parallel. At 3 dpi a significant reduction was observed in the lesion area compared to the control. These results suggest that pectin modification, mediated by VviPME10, plays an important role in the grapevine response to Bc, in particular it seems to behave more like a resistance gene than a susceptibility one. For this reason, it could be considered as a valuable target to improve resistance to Bc in susceptible grapevine varieties. Finally, a molecular method for Bc detection, based on quantitative RT-PCR assays, was set up and applied to estimate the Bc load in field conditions. Although the method allowed the successful detection of the presence of the fungus in samples at different developmental stages, from two V. vinifera cultivars, in different vineyards, the lack of environmental conditions for the development of the disease might have impaired the correlation between detection and the development of the disease. Nonetheless, the method represents a good alternative for monitoring the Bc load in the field at the early season, to predict the BcBR severity at harvest and eventually apply the disease management protocols based on the real need.