Sweet potato chlorotic stunt virus: Studies on viral synergism and suppression of RNA silencing

The studies presented in this thesis aimed to a better understanding of the molecular biology of Sweet potato chlorotic stunt virus (SPCSV, Crinivirus, Closteroviridae) and its role in the development of synergistic viral diseases. The emphasis was on the severe sweet potato virus disease (SPVD) that results from a synergistic interaction of SPCSV and Sweet potato feathery mottle virus (SPFMV, Potyvirus, Potyviridae). SPVD is the most important disease affecting sweetpotato. It is manifested as a significant increase in symptom severity and SPFMV titres. This is accompanied by a dramatic sweetpotato yield reduction. SPCSV titres remain little affected in the diseased plants. Viral synergistic interactions have been associated with the suppression of an adaptive general defence mechanism discovered in plants and known as RNA silencing. In the studies of this thesis two novel proteins (RNase3 and p22) identified in the genome of a Ugandan SPCSV isolate were shown to be involved in suppression of RNA silencing. RNase3 displayed a dsRNA-specific endonuclease activity that enhanced the RNA-silencing suppression activity of p22. Comparative analyses of criniviral genomes revealed variability in the gene content at the 3´end of the genomic RNA1. Molecular analyses of different isolates of SPCSV indicated a marked intraspecific heterogeneity in this region where the p22 and RNase3 genes are located. Isolates of the East African strain of SPCSV from Tanzania and Peru and an isolate from Israel were missing a 767-nt fragment that included the p22 gene. However, regardless of the absence of p22, all SPCSV isolates acted synergistically with SPFMV in co-infected sweetpotato, enhanced SPFMV titres and caused SPVD. These results showed that p22 is dispensable for development of SPVD. The role of RNase3 in SPVD was then studied by generating transgenic plants expressing the RNase3 protein. These plants had increased titres of SPFMV (ca. 600-fold higher in comparison with nontransgenic plants) 2-3 weeks after graft inoculation and displayed the characteristic SPVD symptoms. RNA silencing suppression (RSS) activity of RNase3 was detected in agroinfiltrated leaves of Nicotiana bethamiana. In vitro studies showed that RNase3 was able to cleave small interferring RNAs (siRNA) to products of ~14-nt. The data thus identified RNase3 as a suppressor of RNA silencing able to cleave siRNAs. RNase3 expression alone was sufficient for breaking down resistance to SPFMV in sweetpotato and for the development of SPVD. Similar RNase III-like genes exist in animal viruses which points out a novel and possibly more general mechanism of RSS by viruses. A reproducible method of sweetpotato transformation was used to target RNA silencing against the SPCSV polymerase region (RdRp) with an intron-spliced hairpin construct. Hence, engineered resistance to SPCSV was obtained. Ten out of 20 transgenic events challenged with SPCSV alone showed significantly reduced virus titres. This was however not sufficient to prevent SPVD upon coinfection with SPFMV. Immunity to SPCSV seems to be required to control SPVD and targeting of different SPCSV regions need to be assessed in further studies. Based on the identified key role of RNase3 in SPVD the possibility to design constructs that target this gene might prove more efficient in future studies.

Dancing and Professional Dancers in Roman Egypt

The main aim of the study is to create a many-sided view of dancing in Roman Egypt (1st - early 4th centuries AD) and especially of the dancers who earned their living by dancing as hired performers. Even though dancers and other performers played a central part in many kinds of festivities throughout the ancient world, research on ancient professional dancers is rare and tends to rest on the ancient literature, which reflects the opinions of the elite. Documentary written sources (i.e., papyri, ostraka) the core of the present study are mentioned rather superficially, easily resulting in a stereotypical view of the dancers. This study will balance the picture of professional dancers in antiquity and of ancient dancing in a more general sense. The second aim characterizes this study as basic research: to provide a corpus of written sources from Greco-Roman Egypt on dancing and to discuss pictorial sources contemporary with the texts. The study also takes into account the theoretical discussion that centres on dancing as a nonverbal communicative mode. Dancers are seen as significant conveyors of social and cultural matters. This study shows that dancers were hired to perform especially in religious contexts, where the local associations on the village level also played an important part as the employers of the performers. These performers had a better standard of living in economic terms than the average hired worker, and dancers were better paid than other performers. In the Egyptian villages and towns, where the dancers performed and lived, the dancers do not seem to have been marginal because they were professionals or because of some ethnic or social background. However, their possible marginality may have occurred for reasons related to the practicalities of their profession (e.g., the itinerant life style). The oriental background of performers was a literary topos reflecting partly the situation in the centres of the empire, especially Rome, where many performers were of other than Roman origin. The connection of dancing, prostitution and slavery reflects the essential link between dance, body and gender: dancers are equated with such professions or socio-legal statuses where the body is the focus of attention, a commodity and a source of sensual pleasure; this dimension is clearly observable in ancient literature. According to the Egyptian documentary sources, there is no watertight evidence that professional dancers would have been engaged in prostitution and very little, if any, evidence that the disapproval of the professional dancers expressed by the ancient authors was shared by the Egyptians. From the 4th century onwards the dancers almost disappear from the documentary sources, reflecting the political and religious changes in the Mediterranean east.

Text and Intensification of Its Impact in Chishti Samā`

In my master’s thesis I analyse mystical Islamic poetry in ritualistic performance context, samā` , focusing on the poetry used by the Chishti Sufis. The work is based on both literary sources and ethnographic material collected in India. The central textual source is Surūd-i Rūhānī, a compilation of mystical poetry. Textual sources, however, can be understood properly only in relation to the living performance context and therefore I also utilise interviews of Sufis and performers of mystical music and recordings of samā` assemblies along with texts. First part of the thesis concentrates on thematic overview of the poems and the process of selecting a suitable text for performance. The poems are written in three languages, viz. in Persian, Urdu and Hindi. Among the authors are both Sufis and non-Sufis. The poems, mystical and non-mystical alike, share the same poetic images and they acquire a mystical meaning when they are set to qawwali music and performed in samā` assemblies. My work includes several translations of verses not previously translated. Latter part of the thesis analyses the musical idiom of qawwali and the ways in which the impact of text on listeners is intensified in performance. Typically the intensification is accomplished in the level of a single poem through three different techniques: using introductory verses, inserting verses between the verses of the main poem and repeating individual units of text. The former two techniques are tied to creating a mystical state in the listeners while the latter aims at sustaining it. It is customary that a listener enraptured by mystical experience offers a monetary contribution to the performers. Thus, intensification of the text’s impact aims at enabling the listeners to experience mystical states.

Feeling Better, Performing Better? Holistically-Oriented Top Performance and Well-Being (HOPE) : Performance Enhancement and Its Perceived Impacts on Musicians

Most musicians choose a career in music based on their love of the art and a desire to share it with others. However, being a performing musician is highly demanding. Despite considerable evidence of the great frequency of performance-related problems (e.g. debilitating performance anxiety) among professional musicians or aspiring musicians in the current Western classical music tradition these problems are seldom discussed openly. The existing system offers musicians very little help in learning how to build sustainable performance success into their musical career. This study it is first of its kind in Finland which addresses the issue on larger scale in a systematic way. I devised the HOPE intervention (Holistically-Oriented Top Performance and Well-Being Enhancement), in order to learn how to integrate professional peak performance and a sense of personal well-being into the lives and careers of musicians. Unlike most interventions in previous research, the HOPE intervention is explicitly holistic and aims at enhancing the whole musician, not just alleviating performance anxiety. Earlier research has not in principle focused on musicians´ psychological well-being or on their subjective perceptions. The main purpose of the study is to understand the perceived impacts of the specially devised HOPE intervention on the participants and particularly in four key areas: performing, playing or singing well-being, and overall (performing, playing or singing and well-being combined). Furthermore, it is hoped that a deeper understanding of performers´ development will be gained. The research method is interdisciplinary and mainly qualitative. The primary data consist of a series of linked questionnaires (before and after the intervention) and semi-structured follow-up interviews collected during action research-oriented HOPE intervention courses for music majors in the Sibelius Academy. With the longitudinal group called Hope 1, the core data were collected during a nine month HOPE intervention course and from follow-up interviews conducted six months later in 2003-2004. The core data of Hope 1 (nine participants) are compared with the perceived impacts on fifty-three other participants in the HOPE courses during the period since their inception, 2001-2006. The focus is particularly on participants´ subjective perceptions. Results of the study suggest that the HOPE intervention is beneficial in enhancing overall performance capacity, including music performance, and a personal sense of well-being in a music university setting. The findings indicate that within all key areas significant positive changes take place between the beginning and the end of a HOPE intervention course. The longitudinal data imply that the perceived positive changes are still ongoing six months after the HOPE intervention course is finished. The biggest change takes place within the area of performing and the smallest, in participants´ perception of their playing or singing. The main impacts include reduced feelings of stress and anxiety (an enhanced sense of well-being) as well as increased sense of direction and control in one's life. Since the results of the present research gave no other reason to believe otherwise, it is to be expected that the HOPE intervention and the results of the study can be exploited in other areas of human activity as well, especially where continuous professional top performance is a prerequisite such as in business or sports. Keywords: performance enhancement, professional top performance, subjective well-being, subjective perceptions, holism, coaching, music performance anxiety, studying music, music.

Muusikon mentaaliharjoittelu : Konsertoivan ja opettavan pianistin mentaaliharjoittelu

This research explores the foci, methods and processes of mental training by pianists who are active as performers and teachers. The research is based on the concept of mental training as a solely mental mode of practising. Musician s mental training takes place without an instrument or the physical act of playing. The research seeks answers to questions: 1) What are the foci of a pianist s mental training? 2) How does a pianist carry out the mental training? 3) What does mental training in music entail as a process? The research approach is qualitative, and the materials were gathered from thematic interviews. The aim of practising is always an improved result both in the act of playing and the performance. Mental training by a pianist is collaboration between technical, auditory, visual, kinaesthetic and affective factors. Also interpretation, memory and overcoming stage fright are needed. Technical, cognitive and performance skills are involved. According to the results of this research, mental training is a goal-oriented activity which can have an impact on all of these factors. Without a musical inner ear and its functionality, true musicianship cannot exist. One particular result of this research is the conceptualisation of opening up the inner ear. Auditory exercises and internally playing mental images are essential elements of the mental practice of a musician. Visual images, such as a picture of music notation or a performance event, are the point of focus for musicians who find visual images to be the easiest to realise. When developing technical skills by using mental training, it is important to focus on the technically most difficult sections. It is also necessary to focus on the holistic experiencing of the performance situation. By building on positive energies and strengths, the so-called psyching up may be the most important element in mental training. Based on the results of this research, a synthesis is outlined of the music event as an activity process, built on representations and schemes. Mental training aims at the most ideal possible act of playing and the creation of a musical event; these are achieved by focussing on various mental images produced by the different senses, together with concrete practising. Mental training in sports and in music share common factors. Both modes of practising, mental as well as physical, involve three important elements: planning, realisation and evaluation of the practice. In music, however, the goal is an artistic end result which does not often apply to an athletic event. Keywords: Mental training in music, auditory imagining, visualisation, kinaesthetic-mental experience, mastery of the psyche

The Significance of Wild Plants in the Evolutionary Ecology of Three Major Viruses Infecting Cultivated Sweetpotato in Uganda

The studies presented in this thesis contribute to the understanding of evolutionary ecology of three major viruses threatening cultivated sweetpotato (Ipomoea batatas Lam) in East Africa: Sweet potato feathery mottle virus (SPFMV; genus Potyvirus; Potyviridae), Sweet potato chlorotic stunt virus (SPCSV; genus Crinivirus; Closteroviridae) and Sweet potato mild mottle virus (SPMMV; genus Ipomovirus; Potyviridae). The viruses were serologically detected and the positive results confirmed by RT-PCR and sequencing. SPFMV was detected in 24 wild plant species of family Convolvulacea (genera Ipomoea, Lepistemon and Hewittia), of which 19 species were new natural hosts for SPFMV. SPMMV and SPCSV were detected in wild plants belonging to 21 and 12 species (genera Ipomoea, Lepistemon and Hewittia), respectively, all of which were previously unknown to be natural hosts of these viruses. SPFMV was the most abundant virus being detected in 17% of the plants, while SPMMV and SPCSV were detected in 9.8% and 5.4% of the assessed plants, respectively. Wild plants in Uganda were infected with the East African (EA), common (C), and the ordinary (O) strains, or co-infected with the EA and the C strain of SPFMV. The viruses and virus-like diseases were more frequent in the eastern agro-ecological zone than the western and central zones, which contrasted with known incidences of these viruses in sweetpotato crops, except for northern zone where incidences were lowest in wild plants as in sweetpotato. The NIb/CP junction in SPMMV was determined experimentally which facilitated CP-based phylogenetic and evolutionary analyses of SPMMV. Isolates of all the three viruses from wild plants were genetically similar to those found in cultivated sweetpotatoes in East Africa. There was no evidence of host-driven population genetic structures suggesting frequent transmission of these viruses between their wild and cultivated hosts. The p22 RNA silencing suppressor-encoding sequence was absent in a few SPCSV isolates, but regardless of this, SPCSV isolates incited sweet potato virus disease (SPVD) in sweetpotato plants co-infected with SPFMV, indicating that p22 is redundant for synergism between SCSV and SPFMV. Molecular evolutionary analysis revealed that isolates of strain EA of SPFMV that is largely restricted geographically in East Africa experience frequent recombination in comparison to isolates of strain C that is globally distributed. Moreover, non-homologous recombination events between strains EA and C were rare, despite frequent co-infections of these strains in wild plants, suggesting purifying selection against non-homologous recombinants between these strains or that such recombinants are mostly not infectious. Recombination was detected also in the 5 - and 3 -proximal regions of the SPMMV genome providing the first evidence of recombination in genus Ipomovirus, but no recombination events were detected in the characterized genomic regions of SPCSV. Strong purifying selection was implicated on evolution of majority of amino acids of the proteins encoded by the analyzed genomic regions of SPFMV, SPMMV and SPCSV. However, positive selection was predicted on 17 amino acids distributed over the whole the coat protein (CP) in the globally distributed strain C, as compared to only 4 amino acids in the multifunctional CP N-terminus (CP-NT) of strain EA largely restricted geographically to East Africa. A few amino acid sites in the N-terminus of SPMMV P1, the p7 protein and RNA silencing suppressor proteins p22 and RNase3 of SPCSV were also submitted to positive selection. Positively selected amino acids may constitute ligand-binding domains that determine interactions with plant host and/or insect vector factors. The P1 proteinase of SPMMV (genus Ipomovirus) seems to respond to needs of adaptation, which was not observed with the helper component proteinase (HC-Pro) of SPMMV, although the HC-Pro is responsible for many important molecular interactions in genus Potyvirus. Because the centre of origin of cultivated sweetpotato is in the Americas from where the crop was dispersed to other continents in recent history (except for the Australasia and South Pacific region), it would be expected that identical viruses and their strains occur worldwide, presuming virus dispersal with the host. Apparently, this seems not to be the case with SPMMV, the strain EA of SPFMV and the strain EA of SPCSV that are largely geographically confined in East Africa where they are predominant and occur both in natural and agro-ecosystems. The geographical distribution of plant viruses is constrained more by virus-vector relations than by virus-host interactions, which in accordance of the wide range of natural host species and the geographical confinement to East Africa suggest that these viruses existed in East African wild plants before the introduction of sweetpotato. Subsequently, these studies provide compelling evidence that East Africa constitutes a cradle of SPFMV strain EA, SPCSV strain EA, and SPMMV. Therefore, sweet potato virus disease (SPVD) in East Africa may be one of the examples of damaging virus diseases resulting from exchange of viruses between introduced crops and indigenous wild plant species. Keywords: Convolvulaceae, East Africa, epidemiology, evolution, genetic variability, Ipomoea, recombination, SPCSV, SPFMV, SPMMV, selection pressure, sweetpotato, wild plant species Author s Address: Arthur K. Tugume, Department of Agricultural Sciences, Faculty of Agriculture and Forestry, University of Helsinki, Latokartanonkaari 7, P.O Box 27, FIN-00014, Helsinki, Finland. Email: tugume.arthur@helsinki.fi Author s Present Address: Arthur K. Tugume, Department of Botany, Faculty of Science, Makerere University, P.O. Box 7062, Kampala, Uganda. Email: aktugume@botany.mak.ac.ug, tugumeka@yahoo.com

Diccionario Enciclopédico de Música en Mexico

The work is the most extensive encyclopedic compilation on Mexican music published to the date, in the form of a dictionary. It includes composer's bios, list of works, musical institutions, instruments, performers, theatres, and many other categories about the music of Mexico. Most of entries include bibliography.

Cosmic Clowns : Convention, Invention, and Inversion in the Yaqui Easter Ritual

Cosmic Clowns: Convention, Invention, and Inversion in the Yaqui Easter Ritual is an ethnographic study of masked clown figures called Chapayekas. They represent Judas and the Roman soldiers in the Passion play that forms the narrative core of the Easter ritual of the Yaquis, an indigenous group in Sonora, Mexico. The study looks at how the Chapayeka is created as a ritual figure, how their performance is constructed, and what the part of the clown is in the dynamics of the ritual. The material was gathered over three periods of anthropological fieldwork in Cócorit, Sonora during Easter in 2004, 2006 and 2007. The Chapayeka masks portray foreigners, animals, mythological figures, and even figures from television and movies. They combine two kinds of performance: they perform set, conventional actions, and improvise and invent new ones. This creates dialectics of invention and convention that allow the figure to mediate between the ritual and its context and different kinds of beings within the Yaqui cosmology. The conventional side of their performance is a cycle of death and rebirth that is an inversion of the cycle of Jesus. Through invention, they separate themselves from the other performers and make themselves powerful. Alternation between the two modes enhances that power and brings it into the conventions of the ritual; ultimately the Chapayekas revitalize the entire ritual. The study finds that the clowns are extremely important to the continuity of both ritual and culture, as the combination of continuity and change, convention and invention, is what makes it possible to recreate the conventions of Yaqui culture as powerful and compelling in various contexts. Another factor is the prevalence of dialectical mediation, which relates concepts by defining them against each other as opposites, and makes it possible to cross a boundary while keeping it intact. Clowns embody and create dialectics to mediate boundaries while guarding against relativization, the disappearance of distinctions. The Chapayekas create and constitute boundaries between the self and other, microcosm and macrocosm, sacred and profane. The study argues that all clown and trickster figures are characterized by constantly alternating between invention and convention; this is what connects them to the collective and moral aspect of culture and, at the same time, makes them unpredictable and powerful. It is possible to do justice to the opposed aspects of these ambiguous and paradoxical figures by taking into account the different foundations and contextual effects of the different modes of symbolization.