农杆菌介导的双价抗盐基因转化番茄的研究

近年来，植物耐盐生物技术研究取得了可喜的进展，特别是通过抗盐基因转化在一定程度上使植物的耐盐性得到了提高。然而，植物的耐盐性是一个多基因控制的复杂性状，依赖于多个基因之间的相互作用。因此，只是将单个基因导入植物获得的抗逆性还是远不能达到满意的效果。一般认为，将多个与耐盐相关的基因转入到同一个植物（即所谓的“复合基因转化”）将会大大提高转基因植物的耐盐能力。 渗透调节是植物抵御盐胁迫的主要方式。植物渗透调节的方式分为两类：一是在细胞中吸收和积累无机盐，如通过离子通道、Na+/H+逆向运输蛋白和ATP酶/H+泵;二是在细胞中合成有机溶质，如脯氨酸和甘氨酸甜菜碱。 我们通过农杆菌介导法向转AtNHX1（拟南芥Na+/H+逆向运输蛋白编码基因）的番茄（Lycopersicum esculentum L. ‘Moneymaker’）株系X1OEA1自交二代植株（T2）中转入山菠菜甜菜碱醛脱氢酶基因（BADH）。PCR、Southern、RT-PCR和甜菜碱含量分析结果证明，BADH已经整合到目标植物基因组，并在转基因植株中转录和翻译表达。叶绿素荧光（Fv/Fm）、相对电导率（Rc/Rc’）、叶绿素含量（Chla+b）、叶绿素a/b比（Chla/b）和光合速率（Pn）测定结果表明，在200 mM NaCl 胁迫下，二次转化的番茄植株各项生理指标均优于转单基因AtNHX1的番茄。初步证明“复合基因转化”有助于进一步提高植物的耐盐性。同时对番茄的转化系统进行了优化，结果表明使用抗生素‘特美汀’作为抑制农杆菌的抗生素的转化效率明显高于使用头孢霉素。

Extreme Sports: A positive transformation in courage and humility

Extreme sports and extreme sports participants have been most commonly explored from a negative perspective, for example the “need to take unnecessary risks.” This study explored what can be learned from extreme sports about courage and humility - two positive psychology constructs. A phenomenological method was used via unstructured interviews with 15 extreme sports participants and other first hand accounts. The extreme sports included B.A.S.E. jumping, big wave surfing, extreme skiing, waterfall kayaking, extreme mountaineering and solo rope-free climbing. Results indicate that humility and courage can be deliberately sought out by participating in activities that involve a real chance of death, fear and the realisation that nature in its extreme is far greater and more powerful than humanity.

Creative industries in China : four perspectives on social transformation

In late 2004, the concept of the creative industries arrived in China. It was warmly welcomed in Shanghai then subsequently adopted with some degree of caution in Beijing. In the years since, officials, scholars, practitioners, entrepreneurs and developers have exploited of the idea of creative industries, and a range of associated terms, to construct an alternative vision of an emerging China. In 2009, Li Wuwei, the Director of the Shanghai Creative Industries Association, himself a leading player in national political reform, released a book titled Creativity is Changing China (Chuangyi gaibian Zhongguo), subsequently translated as Creative Industries Are Changing China in English. The paper investigates the uptake of the creative industries in China and asks: can they really change China, or are they just rearranging the cultural landscape in some cities?

Applied theatre and organisational transformation

Applied Theatre is an umbrella term for a range of drama-based techniques, all of which align with a lineage of pedagogical theory and practice: (e.g.) Freire, Moreno, Heathcote. It encompasses methods and forms including Drama Education (O’Neill); Forum Theatre (Boal); and Process Drama (Haseman, O’Toole). Applied theatre often occurs in non-theatrical settings (schools, hospitals, prisons) with the aim of helping participants address issues of local concern. Increasingly, Applied Theatre practices are utilised in the corporate environment. Appied Theatre adopts artistic principles in production, but posits a practical utility beyond simple entertainment.

Dynamics of transformation and fragmentation of composite liquid nano-particles

Recent research on particle size distributions and particle concentrations near a busy road cannot be explained by the conventional mechanisms for particle evolution of combustion aerosols. Specifically they appear to be inadequate to explain the experimental observations of particle transformation and the evolution of the total number concentration. This resulted in the development of a new mechanism based on their thermal fragmentation, for the evolution of combustion aerosol nano-particles. A complex and comprehensive pattern of evolution of combustion aerosols, involving particle fragmentation, was then proposed and justified. In that model it was suggested that thermal fragmentation occurs in aggregates of primary particles each of which contains a solid graphite/carbon core surrounded by volatile molecules bonded to the core by strong covalent bonds. Due to the presence of strong covalent bonds between the core and the volatile (frill) molecules, such primary composite particles can be regarded as solid, despite the presence of significant (possibly, dominant) volatile component. Fragmentation occurs when weak van der Waals forces between such primary particles are overcome by their thermal (Brownian) motion. In this work, the accepted concept of thermal fragmentation is advanced to determine whether fragmentation is likely in liquid composite nano-particles. It has been demonstrated that at least at some stages of evolution, combustion aerosols contain a large number of composite liquid particles containing presumably several components such as water, oil, volatile compounds, and minerals. It is possible that such composite liquid particles may also experience thermal fragmentation and thus contribute to, for example, the evolution of the total number concentration as a function of distance from the source. Therefore, the aim of this project is to examine theoretically the possibility of thermal fragmentation of composite liquid nano-particles consisting of immiscible liquid v components. The specific focus is on ternary systems which include two immiscible liquid droplets surrounded by another medium (e.g., air). The analysis shows that three different structures are possible, the complete encapsulation of one liquid by the other, partial encapsulation of the two liquids in a composite particle, and the two droplets separated from each other. The probability of thermal fragmentation of two coagulated liquid droplets is discussed and examined for different volumes of the immiscible fluids in a composite liquid particle and their surface and interfacial tensions through the determination of the Gibbs free energy difference between the coagulated and fragmented states, and comparison of this energy difference with the typical thermal energy kT. The analysis reveals that fragmentation was found to be much more likely for a partially encapsulated particle than a completely encapsulated particle. In particular, it was found that thermal fragmentation was much more likely when the volume ratio of the two liquid droplets that constitute the composite particle are very different. Conversely, when the two liquid droplets are of similar volumes, the probability of thermal fragmentation is small. It is also demonstrated that the Gibbs free energy difference between the coagulated and fragmented states is not the only important factor determining the probability of thermal fragmentation of composite liquid particles. The second essential factor is the actual structure of the composite particle. It is shown that the probability of thermal fragmentation is also strongly dependent on the distance that each of the liquid droplets should travel to reach the fragmented state. In particular, if this distance is larger than the mean free path for the considered droplets in the air, the probability of thermal fragmentation should be negligible. In particular, it follows form here that fragmentation of the composite particle in the state with complete encapsulation is highly unlikely because of the larger distance that the two droplets must travel in order to separate. The analysis of composite liquid particles with the interfacial parameters that are expected in combustion aerosols demonstrates that thermal fragmentation of these vi particles may occur, and this mechanism may play a role in the evolution of combustion aerosols. Conditions for thermal fragmentation to play a significant role (for aerosol particles other than those from motor vehicle exhaust) are determined and examined theoretically. Conditions for spontaneous transformation between the states of composite particles with complete and partial encapsulation are also examined, demonstrating the possibility of such transformation in combustion aerosols. Indeed it was shown that for some typical components found in aerosols that transformation could take place on time scales less than 20 s. The analysis showed that factors that influenced surface and interfacial tension played an important role in this transformation process. It is suggested that such transformation may, for example, result in a delayed evaporation of composite particles with significant water component, leading to observable effects in evolution of combustion aerosols (including possible local humidity maximums near a source, such as a busy road). The obtained results will be important for further development and understanding of aerosol physics and technologies, including combustion aerosols and their evolution near a source.

The Federal circuit in Bilski : the machine-or-transformation test

In a much anticipated judgment, the Federal Circuit has sought to clarify the standards applicable in determining whether a claimed method constitutes patent-eligible subject matter. In Bilski, the Federal Circuit identified a test to determine whether a patentee has made claims that pre-empt the use of a fundamental principle or an abstract idea or whether those claims cover only a particular application of a fundamental principle or abstract idea. It held that the sole test for determining subject matter eligibility for a claimed process under § 101 is that: (1) it is tied to a particular machine or apparatus, or (2) it transforms a particular article into a different state or thing. The court termed this the “machine-or-transformation test.” In so doing it overruled its earlier State Street decision to the extent that it deemed its “useful, tangible and concrete result” test as inadequate to determine whether an alleged invention recites patent-eligible subject matter.

The transformation of banana with potential virus resistance genes

One approach to reducing the yield losses caused by banana viral diseases is the use of genetic engineering and pathogen-derived resistance strategies to generate resistant cultivars. The development of transgenic virus resistance requires an efficient banana transformation method, particularly for commercially important 'Cavendish' type cultivars such as 'Grand Nain'. Prior to this study, only two examples of the stable transformation of banana had been reported, both of which demonstrated the principle of transformation but did not characterise transgenic plants in terms of the efficiency at which individual transgenic lines were generated, relative activities of promoters in stably transformed plants, and the stability of transgene expression. The aim of this study was to develop more efficient transformation methods for banana, assess the activity of some commonly used and also novel promoters in stably transformed plants, and transform banana with genes that could potentially confer resistance to banana bunchy top nanovirus (BBTV) and banana bract mosaic potyvirus (BBrMV). A regeneration system using immature male flowers as the explant was established. The frequency of somatic embryogenesis in male flower explants was influenced by the season in which the inflorescences were harvested. Further, the media requirements of various banana cultivars in respect to the 2,4-D concentration in the initiation media also differed. Following the optimisation of these and other parameters, embryogenic cell suspensions of several banana (Musa spp.) cultivars including 'Grand Nain' (AAA), 'Williams' (AAA), 'SH-3362' (AA), 'Goldfinger' (AAAB) and 'Bluggoe' (ABB) were successfully generated. Highly efficient transformation methods were developed for both 'Bluggoe' and 'Grand Nain'; this is the first report of microprojectile bombardment transformation of the commercially important 'Grand Nain' cultivar. Following bombardment of embryogenic suspension cells, regeneration was monitored from single transfom1ed cells to whole plants using a reporter gene encoding the green fluorescent protein (gfp). Selection with kanamycin enabled the regeneration of a greater number of plants than with geneticin, while still preventing the regeneration of non-transformed plants. Southern hybridisation confirmed the neomycin phosphotransferase gene (npt II) was stably integrated into the banana genome and that multiple transgenic lines were derived from single bombardments. The activity, stability and tissue specificity of the cauliflower mosaic virus 358 (CaMV 35S) and maize polyubiquitin-1 (Ubi-1) promoters were examined. In stably transformed banana, the Ubi-1 promoter provided approximately six-fold higher p-glucuronidase (GUS) activity than the CaMV 35S promoter, and both promoters remained active in glasshouse grown plants for the six months they were observed. The intergenic regions ofBBTV DNA-I to -6 were isolated and fused to either the uidA (GUS) or gfjJ reporter genes to assess their promoter activities. BBTV promoter activity was detected in banana embryogenic cells using the gfp reporter gene. Promoters derived from BBTV DNA-4 and -5 generated the highest levels of transient activity, which were greater than that generated by the maize Ubi-1 promoter. In transgenic banana plants, the activity of the BBTV DNA-6 promoter (BT6.1) was restricted to the phloem of leaves and roots, stomata and root meristems. The activity of the BT6.1 promoter was enhanced by the inclusion of intron-containing fragments derived from the maize Ubi-1, rice Act-1, and sugarcane rbcS 5' untranslated regions in GUS reporter gene constructs. In transient assays in banana, the rice Act-1 and maize Ubi-1 introns provided the most significant enhancement, increasing expression levels 300-fold and 100-fold, respectively. The sugarcane rbcS intron increased expression about 10-fold. In stably transformed banana plants, the maize Ubi-1 intron enhanced BT6.1 promoter activity to levels similar to that of the CaMV 35S promoter, but did not appear to alter the tissue specificity of the promoter. Both 'Grand Nain' and 'Bluggoe' were transformed with constructs that could potentially confer resistance to BBTV and BBrMV, including constructs containing BBTV DNA-1 major and internal genes, BBTV DNA-5 gene, and the BBrMV coat protein-coding region all under the control of the Ubi-1 promoter, while the BT6 promoter was used to drive the npt II selectable marker gene. At least 30 transgenic lines containing each construct were identified and replicates of each line are currently being generated by micropropagation in preparation for virus challenge.

The role of organisational capacity in the knowledge-based transformation of Brisbane, Australia

Purpose - The cumulative impacts of the knowledge economy together with the emerging dominance of knowledge-intensive sectors, have led to an unprecedented period of socio-economic and spatial restructuring. As a result, the paradigm of knowledge-based urban development (KBUD) has emerged as a development strategy to guide knowledge-based economic transformation (Knight, 1995; Yigitcanlar, 2007). Notwithstanding widespread government commitment and financial investment, in many cases providing the enabling circumstances for KUBUD has proven a complicated task as institutional barriers remain. Researchers and practitioners advocate that the way organisations work and their institutional relationships, policies and programs, will have a significant impact on a regions capacity to achieve KBUD (Savitch, 1998; Savitch and Kantor, 2002; Keast and Mandell, 2009). In this context, building organisational capacity is critical to achieving institutional change and bring together all of the key actors and sources, for the successful development, adoption, and implementation of knowledge-based development of a city (Yigitcanlar, 2009). Design/methodology/approach - There is a growing need to determine the complex inter-institutional arrangements and intra-organisational interactions required to advance urban development within the knowledge economy. In order to design organisational capacity-building strategies, the associated attributes of good capacity must first be identified. The paper, with its appraisal of knowledge-based urban development, scrutinises organisational capacity and institutional change in Brisbane. As part of the discussion of the case study findings, the paper describes the institutional relationships, policies, programs and funding streams, which are supporting KBUD in the region. Originality/value - In consideration that there has been limited investigation into the institutional lineaments required to provide the enabling circumstances for KBUD, the broad aim of this paper is to discover some good organisational capacity attributes, achieved through a case study of Brisbane. Practical implications - It is anticipated that the findings of the case study will contribute to moving the discussion on the complex inter-institutional arrangements and intra-organisaational interactions required for KBUD, beyond a position of rhetoric.

Enhancing agrobacterium-mediated transformation efficiency of sugarcane : progress towards an efficient, genotype-independent method

Several key issues need to be resolved before an efficient and reproducible Agrobacterium-mediated sugarcane transformation method can be developed for a wider range of sugarcane cultivars. These include loss of morphogenetic potential in sugarcane cells after Agrobacterium-mediated transformation, effect of exposure to abiotic stresses during in vitro selection, and most importantly the hypersensitive cell death response of sugarcane (and other nonhost plants) to Agrobacterium tumefaciens. Eight sugarcane cultivars (Q117, Q151, Q177, Q200, Q208, KQ228, QS94-2329, and QS94-2174) were evaluated for loss of morphogenetic potential in response to the age of the culture, exposure to Agrobacterium strains, and exposure to abiotic stresses during selection. Corresponding changes in the polyamine profiles of these cultures were also assessed. Strategies were then designed to minimize the negative effects of these factors on the cell survival and callus proliferation following Agrobacterium-mediated transformation. Some of these strategies, including the use of cell death protector genes and regulation of intracellular polyamine levels, will be discussed.