Conjugal transfer of the symbiotic plasmid in Rhizobium galegae sv. officinalis

Rhizobia are agriculturally important bacteria that possess the ability to fix nitrogen for their host legumes, an attribute ascribed to the presence of symbiosis-related genes usually clustered on plasmids called symbiotic plasmids (pSyms). Many pSyms have been proven self-transmissible, capable of transferring themselves to other bacteria through conjugation, thereby propagating their symbiotic features. Rhizobium galegae symbiovar (sv.) officinalis has a pSym, on which typical conjugation genes have been revealed. A Type IV secretion system (T4SS) functioning as a conjugation system has also been computationally predicted on a chromid, another replicon in R. galegae sv. officinalis. In addition, the transfer of the pSym of R. galegae sv. officinalis to a non-nodulating mutant strain of R. galegae sv. orientalis has been previously observed under laboratory conditions. Therefore, this thesis was aimed at investigating the self-transmissibility of the pSym of R. galegae sv. officinalis and the necessity of the T4SS’ presence for the pSym transfer. Two derivatives of the R. galegae sv. officinalis were generated with one strain cured of its pSym by using Tn5-mob-sacB transposon and the other strain excised the T4SS from the chromid by Cre-lox site specific recombination system. Conjugation were then performed between these two derivatives as well as between the wild-type strain and the plasmid-cured derivative, followed by the host plant nodulation tests. The tests showed no formation of a single nodule in either pair, which was unexpectedly inconsistent with the previous experimental observation. No solid explanations could be proposed at this stage. It might be due to the low transfer frequency resulted from complex associations with subtle environmental signal molecules or recipient cell recognition that presumably disabled the transmissibility of the pSym

Decay Properties Of Multilinear Oscillatory Integrals

In this thesis, we study the following multilinear oscillatory integral introduced by Christ, Li, Tao and Thiele \cite{CLTT} \begin{equation} I_{\lambda}(f_1,...f_n)=\int_{\mathbb{R}^m}e^{i\lambda P(x)}\prod_{j=1}^n f_j(\pi_j(x))\eta(x)dx, \end{equation} where $P:\mathbb{R}^m\to\mathbb{R}$ is a real-valued measurable function, $\eta$ is a compactly supported smooth cutoff function. Each $\pi_j$ is a surjective linear transformation from $\mathbb{R}^m$ to $\mathbb{R}^{k_j}$, where $1\le k_j\le m-1$. Each $f_j:\mathbb{R}^{k_j}\rightarrow \mathbb{C}$ is a locally integrable function with respect to Lebesgue measure on $\mathbb{R}^{k_j}$. In Chapter 2, we first introduce the nondegeneracy degree along with the nondegeneracy norm defined in \cite{CLTT} to characterize the nondegeneracy condition of the phase function. In the same chapter, we will summarize some powerful tools that can help to simplify the problem and introduce the idea of a special geometric structure called ``separation . There are three results in this thesis. The first proves trilinear oscillatory integrals with nondegenerate polynomial phase always have the decay property. The second one extends the one-dimensional case whose phase function has large nondegeneracy degree. The third result deals with the case where every linear mapping preserves the direct sum decomposition

Tan Dun\u27s Eight Memories in Watercolor: Insights into performance

Shostakovich, as· a composer also living under a dictatorship, taught me to express deep humanity. From Takemitsu, I learned that Western and Eastern instruments can be part of the same color palette. John Cage led me to discover structures and sounds as yet unknown, by always keeping an open mind. I owe deepest thanks to these three composers, whose contributions to music have also helped me develop myself---as a composer from a·. traditional culture, growing up in a high-pressure society, living in a now international world. (Tan Dun) The global reach of Tan Dun\u27s music reached its zenith recently at the 2001 Academy Awards where he received an Oscar for the music to the film Crouching Tiger, Hidden Dragon. He is as equally lauded in China- where, in fact, he is their most prominent cultural export- as he is in the West: where he enjoys commissions from the world\u27s finest ensembles including the Metropolitan Opera and many international symphony orchestras. Though particularly known for his long epic-scale music, this dissertation\u27s subject matter centres round Tan Dun as a young 21-year-old, before his successes in the West. In particular, the dissertation is a study of performance practices in Tan\u27s opus one, Eight Memories in Watercolor. Being of similar cultural background to Tan Dun - I was born in the same province as him in China, and educated as a young adult in the West- I am able to offer from the inside a reading of Eight Memories in Watercolor that draws on the performance sensibilities of both Chinese music and western classical music. The dissertation is part exegesis- i.e. it documents how I arrived at an interpretation, and part pedagogical/practical, i.e. it offers a performance practice guide to those pianists seeking to probe some of the Chinese \u27meanings\u27 latent in the music. Chapter one sets the scene, and offers both a background to the place of music in the Cultural Revolution, and a historical sketch of the significance of the Beijing conservatory to the development of Tan Dun\u27s music. Chapter two opens by setting out some parameters of performance practice theory, and in particular how these theories can be used to illuminate Tan\u27s piano music. Chapter three traces the myriad of styles in Eight Memories in Watercolor. Through analysis, I show the work\u27s close relationship to traditional Chinese music and also illustrate aesthetic elements that are drawn from both western and Chinese cultures. This chapter is to be read as a guide to performing this work, and, by dint of sharing the same culturally diverse background as Tan Dun, it is my hope that my reading of his music and that my guide to performance will offer to western pianists a richer starting point from which to craft their performances. I put this theory to the test in chapter four, where I conduct an experiment illustrating how my performance directions can assist non-Chinese pianists in capturing the spirit of Tan Dun\u27s music. The results of this experiment are offered through a transcript and analysis of three interviews, one by each of the pianists undertaking the experiment. The dissertation concludes with an evaluation of the success of my research and offers some areas for further study

A Sandbox Development For Demonstrating Bom Transmission In A Plm And Erp Integrated System

Integrating Product Lifecycle Management (PLM) and Enterprise Resource Planning (ERP) in a company is important. Currently, many companies have difficulties on integrating PLM and ERP systems. This study developed a prototype (sandbox) of PLM and ERP integrated system for demonstrating BOM transmission in a PLM and ERP integrated system, describing key aspects of system integration and BOM transmission. This sandbox will facilitate users\u27 knowledge on how to process transactions in a PLM and ERP integrated system. The BOM accuracy was examined by comparing BOM data in PLM system to BOM data in ERP system after implementing engineering changes. This study was limited to Teamcenter PLM, SAP ERP and Tesis Gateway for SAP

Genome, transcriptome, and methylome in the conifer pathogen Heterobasidion parviporum

The fungus Heterobasidion parviporum is considered as the most destructive disease agent of conifers. The main host of H. parviporum is Norway spruce (Picea abies), and the primary infection is mediated by aerial sexual spores (basidiospores) landing on fresh stump surfaces or tree injuries. The secondary infection is mediated by vegetative spread of the fungus via root contacts formed between neighboring trees. H. parviporum features a dual and flexible lifestyle (saprotrophy and necrotrophy). Due to the unavailability of a reference genome, studies of the molecular pathology have heavily relied on parallel studies on its sibling species H. irregulare. The rapid development of next-generation sequencing techniques has revolutionized the scalability, reliability, and resolution of sequencing in life sciences. The first aim of this study was to provide a reference genome for H. parviporum by application of whole-genome sequencing. We selected a virulent isolate (isolate 96026) as the reference, which presented us a genome assembly of 37.76 Mb, hosting 10,502 protein-coding genes. To identify genomic variations potentially accountable for the virulence of the reference isolate, 14 less virulent isolates were also sequenced. Comparative genomic analysis uncovered not only the remarkable intraspecific level of polymorphism, but also two genomic regions exclusive to the reference isolate. The proteins encoded by genes situated in these two regions included a cytochrome P450, a major facilitator superfamily transporter, and secreted proteins, suggestive of their potential implication in the virulence of H. parviporum reference isolate. To propose candidate virulence factors for functional characterizations, secreted protein-coding genes under genome-wide selection pressure or possessing featured variants were explored, and examples were listed. We then targeted small-secreted proteins (SSPs) in H. parviporum reference isolate to provide promising SSP candidate(s) with functional evidence in terms of promoting disease development. A particular SSP (HpSSP35.8) was found capable of causing rapid cell death on the leaves of Nicotiana benthamiana and of triggering the expression of some defense-related genes in N. benthamiana. In addition, HpSSP35.8 was also highly expressed during the pre-symptomatic phase of H. parviporum infection of Norway spruce seedlings, in which some defense-related genes were also induced. Collectively, all evidence suggests this SSP as a potentially important virulence candidate, presumably in the early stage of pathogenic interaction with the host. Lastly, the transcriptome and DNA methylome profiles of the reference isolate growing as a necrotroph and as a saprotroph were obtained. A group of carbohydrate-active enzymes was found essential for its both necrotrophic and saprotrophic growth, and a small set of signaling- and transcription factors might be associated with the transition of saprotrophic to necrotrophic growth. DNA methylation was demonstrated to mostly target and repress the activity of transposable elements in the genome. A small group of genes with significantly varied methylation and expression levels were identified, implying the potential of DNA methylation as one of the transcriptional regulatory mechanisms in this fungus. Our study provided the reference genome and first methylome of H. parviporum. Follow-up research into the molecular mechanisms of Heterobasidion pathogenesis could be greatly facilitated by the availability of the genome, transcriptome, methylome and secretome data.Kuusentyvilaho (Heterobasidion parviporum) on metsäpatogeeni, joka lahottaa kuusen juuristoa ja runkoa. Kuusentyvilaho sieni tartuttuu ilmalevintäisillä itiöillä, jota kutsutaan kantaitiöiksi. Ilmassa olevat itiöt itävät laskeutuessaan kantoihin tai juuren tai haarojen haavoihin. Sen jälkeen sieni kehittyy ja leviää viereisiin terveisiin puihin juuriyhteyksien kautta. Vaikka H. parviporum nekrotrofina voi tappaa isäntapuita ravinteiden saamiseksi kasvua ja lisääntymistä varten, sieni voi myös suoraan syödä kuolleita puusoluja saprotrofina. Tämän väitöskirjan ensimmäinen tavoite oli sekvensoida tämän sienilajin erittäin virulenttin kannan koko genomi, jotta sitä voitiin käyttää vertailugenomina. Sienen koko genomin koko oli 37,76 Mb, jossa oli 10 502 ennustettua proteiinia koodaavaa geeniä. Saman sienilajin neljätoista vähemmän virulenttista kantaa myös sekvensoitiin ja verrattiin referenssigenomiin. Vertaileva genomianalyysi paljasti kaksi genomin aluetta, jotka löytyivät yksinomaan virulentissa referenssikannasta. Näihin kahteen alueeseen sisältyvien geenien koodaamat proteiinit sisälsivät sytokromi P450:a, joka on merkittävä entsymaattisten proteiinien superperhe, sekä erittyiviä proteiineja Nämä analyysit osoittavat, näiden kahden alueen voivan olla osatekijöitä referenssikannan virulenssiin. H. parviporumin referenssigenomin avulla seulottiin ja tutkittiin edelleen vertailukannan koodaamia pieniä erittyneitä proteiineja (SSP). Erityisen SSP: n (HpSSP35.8) havaittiin kykenevän aiheuttamaan nopean solukuoleman tupakan sukuisessa kasvin (Nicotiana benthamiana) lehdillä ja käynnistämään joitakin puolustukseen liittyvien geenien ilmentymistä. Lisäksi HpSSP35.8 ilmentyi myös voimakkaasti kuusen taimilla H. parviporum -infektion aikana. Taimissa myös indusoitiin joitakin puolustukseen liittyviä geenejä. Tulokset osoittivat yhdessä, että tämä SSP on potentiaalisesti tärkeä virulenssikandidaatti H. parviporum kasvisairauden aiheuttajana. Lopuksi analysoitiin nekrotrofi ja saprotrofi kasvuvaiheissa kasvavan sienen referenssigenomin transkriptio- ja DNA-metylaatio- profiilit. Ryhmä hiilihydraattiaktiivisia entsyymejä havaittiin välttämättömäksi sekä nekrotrofisen, että saprotrofisen kasvun kannalta, sekä havaittiin pienen joukkon signalointi- ja transkriptiotekijöitä, jotka saattavat liittyä sienen siirtymiseen saprotrofisesta kasvusta nekrotrofiseen kasvuun. DNA-metylaation osoitettiin kohdistuvan pääasiassa liikkuvien geneettisten elementtien (transposoni) aktiivisuuden estämiseen genomissa. Tässä sienessä havaittiin lisäksi pieni ryhmä geenejä, joilla oli merkittävästi vaihtelevia metylaatio- ja ilmentymistasoja, mikä viittaa DNA-metylaation mahdollisuuteen yhtenä transkription säätelymekanismeistä. Kuusentyvilaho patogeneesimekanismien jatkotutkimus voisi hyötyä tutkimuksessamme tuotetuista genomi-, transkriptomi-, metylaatio- ja tiedoista