AGROBEST: an efficient Agrobacterium-mediated transient expression method for versatile gene function analyses in Arabidopsis seedlings

Background: Transient gene expression via Agrobacterium-mediated DNA transfer offers a simple and fast method to analyze transgene functions. Although Arabidopsis is the most-studied model plant with powerful genetic and genomic resources, achieving highly efficient and consistent transient expression for gene function analysis in Arabidopsis remains challenging. Results: We developed a highly efficient and robust Agrobacterium-mediated transient expression system, named AGROBEST (Agrobacterium-mediated enhanced seedling transformation), which achieves versatile analysis of diverse gene functions in intact Arabidopsis seedlings. Using β-glucuronidase (GUS) as a reporter for Agrobacterium-mediated transformation assay, we show that the use of a specific disarmed Agrobacterium strain with vir gene pre-induction resulted in homogenous GUS staining in cotyledons of young Arabidopsis seedlings. Optimization with AB salts in plant culture medium buffered with acidic pH 5.5 during Agrobacterium infection greatly enhanced the transient expression levels, which were significantly higher than with two existing methods. Importantly, the optimized method conferred 100% infected seedlings with highly increased transient expression in shoots and also transformation events in roots of ~70% infected seedlings in both the immune receptor mutant efr-1 and wild-type Col-0 seedlings. Finally, we demonstrated the versatile applicability of the method for examining transcription factor action and circadian reporter-gene regulation as well as protein subcellular localization and protein–protein interactions in physiological contexts. Conclusions: AGROBEST is a simple, fast, reliable, and robust transient expression system enabling high transient expression and transformation efficiency in Arabidopsis seedlings. Demonstration of the proof-of-concept experiments elevates the transient expression technology to the level of functional studies in Arabidopsis seedlings in addition to previous applications in fluorescent protein localization and protein–protein interaction studies. In addition, AGROBEST offers a new way to dissect the molecular mechanisms involved in Agrobacterium-mediated DNA transfer

Characterization of a Bacillus anthracis spore coat-surface protein that influences coat-surface morphology

Bacterial spores are encased in a multilayered proteinaceous shell, called the coat. In many Bacillus spp., the coat protects against environmental assault and facilitates germination. In Bacillus anthracis , the spore is the etiological agent of anthrax, and the functions of the coat likely contribute to virulence. Here, we characterize a B. anthracis spore protein, called CotΒ, which is encoded only in the genomes of the Bacillus cereus group. We found that CotΒ is synthesized specifically during sporulation and is assembled onto the spore coat surface. Our analysis of a cotΒ null mutant in the Sterne strain reveals that CotΒ has a role in determining coat-surface morphology but does not detectably affect germination. In the fully virulent Ames strain, a cotΒ null mutation has no effect on virulence in a murine model of B. anthracis infection.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72138/1/j.1574-6968.2008.01380.x.pd

Small heat-shock protein HspL is induced by VirB protein(s) and promotes VirB/D4-mediated DNA transfer in Agrobacterium tumefaciens

Agrobacterium tumefaciens is a Gram-negative plant-pathogenic bacterium that causes crown gall disease by transferring and integrating its transferred DNA (T-DNA) into the host genome. We characterized the chromosomally encoded alpha-crystallin-type small heat-shock protein (α-Hsp) HspL, which was induced by the virulence (vir) gene inducer acetosyringone (AS). The transcription of hspL but not three other α-Hsp genes (hspC, hspAT1, hspAT2) was upregulated by AS. Further expression analysis in various vir mutants suggested that AS-induced hspL transcription is not directly activated by the VirG response regulator but rather depends on the expression of VirG-activated virB genes encoding components of the type IV secretion system (T4SS). Among the 11 virB genes encoded by the virB operon, HspL protein levels were reduced in strains with deletions of virB6, virB8 or virB11. VirB protein accumulation but not virB transcription levels were reduced in an hspL deletion mutant early after AS induction, implying that HspL may affect the stability of individual VirB proteins or of the T4S complex directly or indirectly. Tumorigenesis efficiency and the VirB/D4-mediated conjugal transfer of an IncQ plasmid RSF1010 derivative between A. tumefaciens strains were reduced in the absence of HspL. In conclusion, increased HspL abundance is triggered in response to certain VirB protein(s) and plays a role in optimal VirB protein accumulation, VirB/D4-mediated DNA transfer and tumorigenesis

Musical decolonisation: the reception of Pablo Casals in Japan and the Sinophone world

This thesis, ‘Musical decolonisation: The reception of Pablo Casals in Japan and the Sinophone world,’ discusses the reception of Western art music in Japan, Taiwan, China, and Hong Kong through the lens of colonialism, the Cold War, and capitalism with a case study on the Spanish cellist and humanitarian Pablo Casals (1876-1973). From the late nineteenth century onwards, Western art music has rooted in Japan and the Sinophone world alongside colonialism. However, postcolonial criticisms of Western art music by scholars from this region were absent until the twenty-first century. To the present day, Western art music enjoys a privileged status in this region as it receives more financial support from both governments and private sectors than local genres. My thesis considers that the phenomenon of Western art music being regarded as superior over local genres was an outcome of cultural imagination under colonisation. The core questions explored in this thesis are: 1) Why was Western art music welcome despite its association with colonialism? 2) Why did not postcolonial criticisms of Western art music gain a foothold in Japan and the Sinophone world in the twentieth century? and 3) What did the consumption of Western art music mean in these places? In this thesis, I argue that the development of postcolonial discourses of Western art music in Japan, Taiwan, China, and Hong Kong was held back by the Cold War and consumer capitalism. In order to gain a clearer understanding of how the development of Western art music in these places was mediated by these factors, I have presented the reception history of Pablo Casals as a case study. Although Casals only visited Japan once, in 1961, and did not have any direct connection with China, Taiwan, and Hong Kong, his name became well-known in the circle of Western art music lovers in these places through written materials and recordings. I have illustrated how Casals was introduced into Japan, Taiwan, and China in the first two decades of the Cold War, how his image was utilised to advance the governmental propaganda of Cold War ideologies, including humanitarianism, anti-communism, and leftism, and how his Cold War image was used to promote his CD sales in the post-Cold War era. Methodologically, by conducting a dialogue with Chen Kuan-Hsing’s ‘inner Asian referential framework,’ my thesis has demonstrated how Casals’ reception in Japan and the Sinophone world served as the commonality amongst these territories to validate Chen’s framework. The research has also further addressed the role of capitalism in Chen’s discussion of the entangled ideologies of colonialism and the Cold War. Drawing further from Chen’s framework, which emphasises the importance of finding an alternative point of reference other than Western academia for knowledge production, my thesis has cited criticisms of Casals’ legacies made by local musicians during his residency from 1957 to 1973 from an indigenous perspective of Puerto Rico. The aim of including the Puerto Rican perspective is to prompt further discussions of musical decolonisation amongst scholars from different regions of the world.</p

Musical decolonisation: the reception of Pablo Casals in Japan and the Sinophone world

This thesis, ‘Musical decolonisation: The reception of Pablo Casals in Japan and the Sinophone world,’ discusses the reception of Western art music in Japan, Taiwan, China, and Hong Kong through the lens of colonialism, the Cold War, and capitalism with a case study on the Spanish cellist and humanitarian Pablo Casals (1876-1973). From the late nineteenth century onwards, Western art music has rooted in Japan and the Sinophone world alongside colonialism. However, postcolonial criticisms of Western art music by scholars from this region were absent until the twenty-first century. To the present day, Western art music enjoys a privileged status in this region as it receives more financial support from both governments and private sectors than local genres. My thesis considers that the phenomenon of Western art music being regarded as superior over local genres was an outcome of cultural imagination under colonisation. The core questions explored in this thesis are: 1) Why was Western art music welcome despite its association with colonialism? 2) Why did not postcolonial criticisms of Western art music gain a foothold in Japan and the Sinophone world in the twentieth century? and 3) What did the consumption of Western art music mean in these places? In this thesis, I argue that the development of postcolonial discourses of Western art music in Japan, Taiwan, China, and Hong Kong was held back by the Cold War and consumer capitalism. In order to gain a clearer understanding of how the development of Western art music in these places was mediated by these factors, I have presented the reception history of Pablo Casals as a case study. Although Casals only visited Japan once, in 1961, and did not have any direct connection with China, Taiwan, and Hong Kong, his name became well-known in the circle of Western art music lovers in these places through written materials and recordings. I have illustrated how Casals was introduced into Japan, Taiwan, and China in the first two decades of the Cold War, how his image was utilised to advance the governmental propaganda of Cold War ideologies, including humanitarianism, anti-communism, and leftism, and how his Cold War image was used to promote his CD sales in the post-Cold War era. Methodologically, by conducting a dialogue with Chen Kuan-Hsing’s ‘inner Asian referential framework,’ my thesis has demonstrated how Casals’ reception in Japan and the Sinophone world served as the commonality amongst these territories to validate Chen’s framework. The research has also further addressed the role of capitalism in Chen’s discussion of the entangled ideologies of colonialism and the Cold War. Drawing further from Chen’s framework, which emphasises the importance of finding an alternative point of reference other than Western academia for knowledge production, my thesis has cited criticisms of Casals’ legacies made by local musicians during his residency from 1957 to 1973 from an indigenous perspective of Puerto Rico. The aim of including the Puerto Rican perspective is to prompt further discussions of musical decolonisation amongst scholars from different regions of the world

The Uncertain Identity of Chinese Music: Symphonization and Adaptation from Western Music in the Development of Taipei Chinese Orchestra

在二十世紀初期，為了追求中國音樂的現代化，中國知識分子及音樂家們開始將中國傳統樂器依據西方交響樂團的架構組成現代國樂團。隨著戰爭造成的移民潮，現代國樂團被傳播到東亞各地區。在目前東亞不同的社會脈絡中，現代國樂團在台灣、香港、中國大陸、新加坡等地有了不同的發展及名稱。目前這種合奏形式在台灣依舊稱國樂；在香港稱中樂；在中國大陸稱民樂、在新加坡則稱華樂。隨著70年代末期台、港兩地的大型國(中)樂團陸續成立；中國大陸的民樂團在文革的結束後重新恢復運作，以及80年代問世的一批大型國(中、民)樂團合奏作品，1980-90年代，國(中、民)樂交響化的討論在台、港、中三地作曲家、指揮家、演奏家間熱烈進行著。在這些討論中，「交響化」一詞成為整個西方音樂文化的縮影，並被視為一個與中國音樂文化相互參照的討論對象，兩岸三地的樂界人士藉由這些討論嘗試為國(中、民)樂團確立未來的發展方向。在交響化的討論中，大陸民樂家企圖建立「具有中國特色的現代民族管弦樂團」；香港則試圖尋找有別於追尋西方音樂美學的「中國式交響化」；台灣學者則開始提出交響化的國樂團，導致了國樂主體性的旁落，而國樂主體性一詞在2000年以後也逐漸成為台灣國樂圈在討論後續發展方向時所使用的詞彙。 然而何謂「國樂主體性」在現今的說法中卻僅及於交響化的國樂團不利於國樂主體性之建立，而缺乏更詳細的論述及完滿的答覆。因此，本文擬以台北市立國樂團歷史發展中與交響化討論最直接相關的西樂中奏作品為例，論述在北市國近年的代表性製作中「國樂主體性」面臨著與台灣國家認同類似的困境。In pursuing musical modernization, Chinese musicians in the early 20th Century began their projects by using Chinese instruments to replicate the European orchestras, thereby forming a contemporary Chinese orchestra. Subsequently, music of the contemporary Chinese orchestra was promoted as “Gouyue,” meaning “national music” in Chinese. After the Sino-Japanese War and the Chinese Civil War, the Chinese Orchestra was disseminated throughout the world as a result of large-scale emigrations. Currently among Asian countries, there are Chinese orchestras in China, Hong Kong, Singapore, and Taiwan; yet, occasionally they are named differently. In China, the genre was renamed to “Minyue,” meaning “ethnic music.” In Singapore the orchestra is referred to as “Huayue” (Chinese music); while in Hong Kong the term “Gongyue” that was used before the 1970s, was later replaced by “Zhongyue” (also meaning “Chinese music”). The name “Gouyue” is now only preserved in Taiwan. At the end of 1970’s, professional Chinese orchestras were established in Taiwan, Hong Kong, and mainland China. At the same time, a significant number of works requiring well-organized Chinese orchestras were composed, which triggered a debate on Chinese Music Symphonization (Guo Yue Jiao Xiang Hua) among the musicians and scholars from these same three places. In the discussions, musicians and scholars from Taiwan, Hong Kong, and mainland China tried to clarify the future of contemporary Chinese orchestra. Participants form mainland China wanted to establish a “Chinese orchestra which contain[ed] Chinese musical features”; Hong Kong sought for a “Chinese model of Symphonization” that was different from Western musical standards; while beginning in the 2000’s, Taiwanese scholars have argued that symphonized Chinese orchestras were disadvantageous towards the establishment of a Gouyue identity. However, “Gou Yue’s identity” has not been well-discussed among the scholars who were making this argument. Hence, this article intends to clarify this question by arguing that the establishment of musical identity is related to the choice of traditional musical elements. Furthermore, Xi Yue Zhong Zou (Western work performed by Chinese orchestra) pieces produced by the Taipei Chinese Orchestra will be taken as examples to demonstrate that “Gou Yue’s identity” experienced a dilemma which is very much akin to the one that the Taiwanese national identity has been experiencing in recent years

李鴻章政術憑藉之機緣網路

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Structure and Process in the Development of Political Thought in Late Ch'ing China: 1842-1911

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