Relevant bibliographies by topics / Novel hybrid approach

Academic literature on the topic 'Novel hybrid approach'

Author: Grafiati
Published: 10 December 2022
Last updated: 19 February 2023

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Journal articles on the topic "Novel hybrid approach"

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Naga Raju, D. V., and V. Vallikumari. "A Novel Hybrid Cryptosystem Based Approach for Secure Multicasting for Dynamic Groups." International Journal of Engineering and Technology 2, no. 4 (2010): 345–49. http://dx.doi.org/10.7763/ijet.2010.v2.145.

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Navarra, Giuseppe, Letterio Rando, Giuseppe La Malfa, Giuseppe Bartolotta, and Giuseppe Pracanica. "Hybrid transvaginal cholecystectomy: a novel approach." American Journal of Surgery 197, no. 6 (June 2009): e69-e72. http://dx.doi.org/10.1016/j.amjsurg.2008.07.059.

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Kumar, Manoj, Sangeet Sriastava, and Arnold Hensman. "A Hybrid Novel Approach of Video Watermarking." International Journal of Signal Processing, Image Processing and Pattern Recognition 9, no. 11 (October 31, 2016): 365–76. http://dx.doi.org/10.14257/ijsip.2016.9.10.35.

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Fang, Fen, Liyuan Li, Ying Gu, Hongyuan Zhu, and Joo-Hwee Lim. "A novel hybrid approach for crack detection." Pattern Recognition 107 (November 2020): 107474. http://dx.doi.org/10.1016/j.patcog.2020.107474.

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Jain, Divya, and Vijendra Singh. "A Novel Hybrid Approach for Chronic Disease Classification." International Journal of Healthcare Information Systems and Informatics 15, no. 1 (January 2020): 1–19. http://dx.doi.org/10.4018/ijhisi.2020010101.

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A two-phase diagnostic framework based on hybrid classification for the diagnosis of chronic disease is proposed. In the first phase, feature selection via ReliefF method and feature extraction via PCA method are incorporated. In the second phase, efficient optimization of SVM parameters via grid search method is performed. The proposed hybrid classification approach is then tested with seven popular chronic disease datasets using a cross-validation method. Experiments are then conducted to evaluate the presented classification method vis-à-vis four other existing classifiers that are applied on the same chronic disease datasets. Results show that the presented approach reduces approximately 40% of the extraneous and surplus features with substantial reduction in the execution time for mining all datasets, achieving the highest classification accuracy of 98.5%. It is concluded that with the presented approach, excellent classification accuracy is achieved for each chronic disease dataset while irrelevant and redundant features may be eliminated, thereby substantially reducing the diagnostic complexity and resulting computational time.
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Khan, Sajid Ullah, Najeeb Ullah, Imran Ahmed, Wang Yin Chai, and Shahid Ullah. "A Novel Hybrid Approach for Medical Image Compression." Current Medical Imaging Reviews 14, no. 6 (October 31, 2018): 845–52. http://dx.doi.org/10.2174/1573405613666170428162650.

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Bahl, Dushyant, Vaibhav Kain, Akshay Sharma, and Mugdha Sharma. "A novel hybrid approach towards movie recommender systems." Journal of Statistics and Management Systems 23, no. 6 (July 29, 2020): 1049–58. http://dx.doi.org/10.1080/09720510.2020.1799503.

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Wang, Jujie, Wenyu Zhang, Jianzhou Wang, Tingting Han, and Lingbin Kong. "A novel hybrid approach for wind speed prediction." Information Sciences 273 (July 2014): 304–18. http://dx.doi.org/10.1016/j.ins.2014.02.159.

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Sekhar, U. V. Chandra, N. Sai sunitha, and M. Prasanna. "A Novel Data Hiding Technique with Hybrid Approach." International Journal of Engineering Trends and Technology 10, no. 4 (April 25, 2014): 205–8. http://dx.doi.org/10.14445/22315381/ijett-v10p239.

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Kannan, Devika, Hassan Mina, Saeede Nosrati-Abarghooee, and Ghasem Khosrojerdi. "Sustainable circular supplier selection: A novel hybrid approach." Science of The Total Environment 722 (June 2020): 137936. http://dx.doi.org/10.1016/j.scitotenv.2020.137936.

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Dissertations / Theses on the topic "Novel hybrid approach"

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Horne, Simon James. "A novel hybrid approach to modeling chemical processes." Thesis, University of Newcastle Upon Tyne, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.531756.

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Hodsgon, Jeremy. "A novel approach to systematically deriving hybrid system models for hydraulic systems." Thesis, University of Bristol, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.500431.

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Turnbull, Julian. "A novel approach to reduce the computation time for CFD : hybrid LES-RANS modelling on parallel computers." Thesis, Cranfield University, 2003. http://hdl.handle.net/1826/104.

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Large Eddy Simulation is a method of obtaining high accuracy computational results for modelling fluid flow. Unfortunately it is computationally expensive limiting it to users of large parallel machines. However, it may be that the use of LES leads to an over-resolution of the problem because the bulk of the computational domain could be adequately modelled using the Reynolds averaged approach. A study has been undertaken to assess the feasibility, both in accuracy and computational efficiency of using a parallel computer to solve both LES and RANS type turbulence models on the same domain for the problem flow over a circular cylinder at Reynolds number 3 900 To do this the domain has been created and then divided into two sub-domains, one for the LES model and one for the kappa - epsilon turbulence model. The hybrid model has been developed specifically for a parallel computing environment and the user is able to allocate modelling techniques to processors in a way which enables expansion of the model to any number of processors. Computational experimentation has shown that the combination of the Smagorinsky model can be used to capture the vortex shedding from the cylinder and the information successfully passed to the kappa - epsilon model for the dissipation of the vortices further downstream. The results have been compared to high accuracy LES results and with both kappa - epsilon and Smagorinsky LES computations on the same domain. The hybrid models developed compare well with the Smagorinsky model capturing the vortex shedding with the correct periodicity. Suggestions for future work have been made to develop this idea further, and to investigate the possibility of using the technology for the modelling of mixing and fast chemical reactions based on the more accurate prediction of the turbulence levels in the LES sub-domain.
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Kadličko, Stephanie Rose. "Evaluation of Serbian commercial corn hybrid tolerance to feeding by larval western corn rootworm (Diabrotica virgifera virgifera LeConte) using the novel difference approach." [Ames, Iowa : Iowa State University], 2008.

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Schneider, Jakob [Verfasser], Paolo Akademischer Betreuer] Carloni, and Marc [Akademischer Betreuer] [Spehr. "Towards pharmaceutical applications on low resolution models of human G protein-coupled receptors: a novel hybrid molecular mechanics/coarse-grained simulation approach / Jakob Schneider ; Paolo Carloni, Marc Spehr." Aachen : Universitätsbibliothek der RWTH Aachen, 2020. http://d-nb.info/1225878810/34.

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Schneider, Jakob Verfasser], Paolo [Akademischer Betreuer] Carloni, and Marc [Akademischer Betreuer] [Spehr. "Towards pharmaceutical applications on low resolution models of human G protein-coupled receptors: a novel hybrid molecular mechanics/coarse-grained simulation approach / Jakob Schneider ; Paolo Carloni, Marc Spehr." Aachen : Universitätsbibliothek der RWTH Aachen, 2020. http://d-nb.info/1225878810/34.

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Wang, Yongchang. "Novel Approaches in Structured Light Illumination." UKnowledge, 2010. http://uknowledge.uky.edu/gradschool_diss/116.

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Among the various approaches to 3-D imaging, structured light illumination (SLI) is widely spread. SLI employs a pair of digital projector and digital camera such that the correspondences can be found based upon the projecting and capturing of a group of designed light patterns. As an active sensing method, SLI is known for its robustness and high accuracy. In this dissertation, I study the phase shifting method (PSM), which is one of the most employed strategy in SLI. And, three novel approaches in PSM have been proposed in this dissertation. First, by regarding the design of patterns as placing points in an N-dimensional space, I take the phase measuring profilometry (PMP) as an example and propose the edge-pattern strategy which achieves maximum signal to noise ratio (SNR) for the projected patterns. Second, I develop a novel period information embedded pattern strategy for fast, reliable 3-D data acquisition and reconstruction. The proposed period coded phase shifting strategy removes the depth ambiguity associated with traditional phase shifting patterns without reducing phase accuracy or increasing the number of projected patterns. Thus, it can be employed for high accuracy realtime 3-D system. Then, I propose a hybrid approach for high quality 3-D reconstructions with only a small number of illumination patterns by maximizing the use of correspondence information from the phase, texture, and modulation data derived from multi-view, PMP-based, SLI images, without rigorously synchronizing the cameras and projectors and calibrating the device gammas. Experimental results demonstrate the advantages of the proposed novel strategies for 3-D SLI systems.
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Liu, Jiamin. "Novel approaches to container loading : from heuristics to hybrid tabu search." Thesis, University of Bedfordshire, 2008. http://hdl.handle.net/10547/295765.

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This work investigates new approaches to the container loading problem which address the issue of how to load three-dimensional, rectangular items (e.g. boxes) into the container in such a way that maximum utilisation is made of the container space. This problem occurs in several industry sectors where the loading approach places cargo effectively into aeroplanes, ships, trailers or trucks in order to save considerable cost. In carrying out this work, the investigation starts by developing a new heuristic approach to the two-dimensional bin packing problem, which has lower complexity than container loading in the aspects of constraints and geometry. A novel approach, including the heuristic strategies and handling method for remaining areas, is developed that can produce good results when testing with benchmark and real world data. Based on the research for two-dimensional bin packing, a novel heuristic approach is developed to deal with the container loading problem with some practical constraints. The heuristic approach to container loading also includes heuristic strategies and the handling of remaining spaces. The heuristic strategies construct effective loading arrangements where combinations of identical or different box types are loaded in blocks. The handling method for remaining spaces further improves the loading arrangements through the representation, partitioning and merging of remaining spaces. The heuristic approach obtains better volume utilisation and the highest stability compared with other published heuristic approaches. However, it does not achieve as high a volume utilisation as metaheuristic approaches, e.g. genetic algorithms and tabu search.To improve volume utilisation, a new hybrid heuristic approach to the container loading problem is further developed based on the tabu search technique which covers the encoding, evaluation criterion and configuration of neighbourhood and candidate solutions. The heuristic strategies as well as the handling method for remaining spaces developed in the heuristic approach are used in this new hybrid tabu search approach. It is shown that the hybrid approach has better volume utilisation than the published approaches under the condition that all loaded boxes with one hundred per cent support from below. In addition, the experimental results show that both the heuristic and hybrid tabu search approaches can also be applied to the multiple container loading problem.
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Dziuda, Łukasz. "Novel approaches to hybrid voltage and current sensing for condition monitoring of remotely operated electrical plant." Thesis, University of Strathclyde, 2007. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.442134.

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Koßmann, Simone [Verfasser]. "Efficient Novel Approaches for the Calculation of Molecular Response Properties : Second-Order Many-Body Perturbation and Double-Hybrid Density Functional Theory / Simone Koßmann." Bonn : Universitäts- und Landesbibliothek Bonn, 2012. http://d-nb.info/1044082984/34.

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Books on the topic "Novel hybrid approach"

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Smith, Holly M. Conclusion. Oxford University Press, 2018. http://dx.doi.org/10.1093/oso/9780199560080.003.0014.

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Chapter 14 provides a summary of the book, describing the epistemic problem in morality, the rationales for seeking usability, and the three salient approaches to solving this problem (the Austere, Pragmatic, and Hybrid solutions). It assesses the strengths and weaknesses of each of these solutions as an approach to the problem of error and the problem of uncertainty. The book introduces a fully-worked out, novel version of the Hybrid solution—termed the “Constrained Standards Hybrid” solution—which is appraised in Chapter 13 as the best available moral system for accommodating the epistemic frailties human decision makers bring to their choices of how to act.
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Morrison, Alan R., Joseph C. Wu, and Mehran M. Sadeghi. Cardiovascular Molecular Imaging. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199392094.003.0029.

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Cardiovascular molecular imaging is a relatively young but rapidly expanding discipline that consists of a biologically-targeted approach to the assessment of physiologic and pathologic processes in vivo. This novel approach to imaging involves the integration of multiple disciplines such as cell and molecular biology, chemistry, and imaging sciences. The ultimate goal is quantitative assessment of cardiovascular processes at the cellular and molecular level, moving beyond traditional diagnostic information, in order to guide individually tailored therapy. In fact, it is likely that specific approaches to molecular imaging will be developed in tandem with the development of novel therapeutic strategies. Recent advances in probe development and imaging systems have contributed to evolution of molecular imaging toward clinical translational. These include technological progress in traditional imaging platforms; along with the emergence of newer imaging modalities such as photoacoustic imaging. In addition, hybrid imaging (e.g. nuclear imaging with CT or MRI) has the potential for improved spatial localization, and more accurate quantification by coupling anatomic and biological information. In addition to potential clinical applications that address existing diagnostic gaps in cardiovascular medicine, molecular imaging allows for unique approaches to studying pathophysiology. This chapter is intended to provide an overview of the state of the art in cardiovascular molecular imaging, highlighting how it may improve the management of major cardiovascular diseases.
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Özkazanç-Pan, Banu. Transnational Migration and the New Subjects of Work. Policy Press, 2019. http://dx.doi.org/10.1332/policypress/9781529204544.001.0001.

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This book brings about insights and key concepts from the field of transnational migration studies to bear upon the field of organization studies. It expands upon multiscalar global perspective, moving beyond methodological nationalism, and historical global conjuncturesas relevant transnational concepts for studying people and difference in novel ways including agentic, reflexive mobile subjectivities as the new subjects of diversity research that emerge in a ‘post-identitarian’ world. Specifically, the book offers transmigrant, hybrid, and cosmopolitan subjectivities as new the subjects of diversity research. Beyond new subjectivities, mobility ontology requires rethinking the epistemology of multiculturalism, examining inequalities, and redirecting the methodologies adopted to attend to difference. In expanding on these, the book offers new frameworks for the study of people on-the-move and organizations through a mobility ontology that foregrounds movement as the natural order of the social world. It also calls into question the ways existing research paradigms and approaches have potentially replicated the creation of boundaries and borders through implicit assumptions about difference, race/ethnicity and belonging. By shifting the ontological premise upon which the field of organization studies rests, this book provides novel ways of theorizing difference, people and work beyond static epistemologies guiding much of the field.
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Imlay, Talbot. The Practice of Socialist Internationalism. Oxford University Press, 2017. http://dx.doi.org/10.1093/oso/9780199641048.001.0001.

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The Practice of Socialist Internationalism examines the efforts of British, French, and German socialist parties to cooperate with one another on concrete international issues. Drawing on archival research in twelve countries, it spans the years from the First World War to the early 1960s, paying particular attention to the two post-war periods (1918 to the late 1920s and 1945 to the mid-1950s), during which national and international politics were recast. During these years, European socialists operated simultaneously in national and transnational spaces, and the book explores the ways in which these two spaces overlapped. In addition to highlighting a neglected dimension of twentieth-century European socialism, it provides novel perspectives on two related subjects: the history of internationalism and the history of international politics. Scholars of internationalism focus either on state or on non-state actors (INGOs), but socialist parties constituted something of a hybrid: rooted more firmly in national politics than most INGOs, they were also more self-consciously internationalist than state actors. Just as importantly, European socialists sought to forge a new practice of international relations, one that would emerge from their collective efforts to work out ‘socialist’ approaches to pressing issues of European politics such as post-war reconstruction, European integration, and decolonization. While the extent of their success is debatable, the efforts of European socialists to identify distinct approaches act as a spotlight, illuminating obscure yet vital aspects of an issue.
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Stegenga, Jacob. Medical Nihilism. Oxford University Press, 2018. http://dx.doi.org/10.1093/oso/9780198747048.001.0001.

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This book defends medical nihilism, which is the view that we should have little confidence in the effectiveness of medical interventions. If we consider the frequency of failed medical interventions, the extent of misleading evidence in medical research, the thin theoretical basis of many interventions, and the malleability of empirical methods in medicine, and if we employ our best inductive framework, then our confidence in the effectiveness of medical interventions ought to be low. Part I articulates theoretical and conceptual groundwork, which offers a defense of a hybrid theory of disease, which forms the basis of a novel account of effectiveness, and this is applied to pharmacological science and to issues such as medicalization. Part II critically examines details of medical research. Even the very best methods in medical research, such as randomized trials and meta-analyses, are malleable and suffer from various biases. Methods of measuring the effectiveness of medical interventions systematically overestimate benefits and underestimate harms. Part III summarizes the arguments for medical nihilism and what this position entails for medical research and practice. To evaluate medical nihilism with care, the argument is stated in formal terms. Medical nihilism suggests that medical research must be modified, that clinical practice should be less aggressive in its therapeutic approaches, and that regulatory standards should be enhanced.
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Book chapters on the topic "Novel hybrid approach"

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Naik, Bighnaraj, Debasmita Mishra, Janmenjoy Nayak, Danilo Pelusi, and Ajith Abraham. "Perturbation Based Efficient Crow Search Optimized FLANN for System Identification: A Novel Approach." In Hybrid Intelligent Systems, 204–12. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-76351-4_21.

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Agarwal, Kavishi, Akshita Sinha, and M. Hima Bindu. "A Novel Hybrid Approach to N-Queen Problem." In Advances in Intelligent and Soft Computing, 519–27. Berlin, Heidelberg: Springer Berlin Heidelberg, 2012. http://dx.doi.org/10.1007/978-3-642-30157-5_52.

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Xue, Xiang, Dong-mei Zhao, Huan-huan Dong, Jin-yi Wang, and Wen-hui Zhao. "A Novel Hybrid Approach for Wind Power Forecasting." In Lecture Notes in Electrical Engineering, 1019–27. New York, NY: Springer New York, 2013. http://dx.doi.org/10.1007/978-1-4614-4981-2_110.

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Weber, Ralf. "Radio Channel Measurement Using a Novel Hybrid Simulation Approach." In Digital Signal Processing for Communication Systems, 93–102. Boston, MA: Springer US, 1997. http://dx.doi.org/10.1007/978-1-4615-6119-4_11.

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Dhruv, Ajay, Aastha Kamath, Anuja Powar, and Karan Gaikwad. "Artist Recommendation System Using Hybrid Method: A Novel Approach." In Emerging Research in Computing, Information, Communication and Applications, 527–42. Singapore: Springer Singapore, 2019. http://dx.doi.org/10.1007/978-981-13-5953-8_44.

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Huang, Jinjie, Ning Lv, and Wenlong Li. "A Novel Feature Selection Approach by Hybrid Genetic Algorithm." In Lecture Notes in Computer Science, 721–29. Berlin, Heidelberg: Springer Berlin Heidelberg, 2006. http://dx.doi.org/10.1007/978-3-540-36668-3_76.

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Redekar, Pooja, and Madhumita Chatterjee. "A Novel Hybrid Approach for Detection of DDoS Attack." In International Conference on Intelligent Data Communication Technologies and Internet of Things (ICICI) 2018, 251–55. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-030-03146-6_27.

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Meselhi, Mohamed A., Hitham M. Abo Bakr, Ibrahim Ziedan, and Khaled Shaalan. "A Novel Hybrid Approach to Arabic Named Entity Recognition." In Communications in Computer and Information Science, 93–103. Berlin, Heidelberg: Springer Berlin Heidelberg, 2014. http://dx.doi.org/10.1007/978-3-662-45701-6_9.

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Agrawal, Uttkarsh, and Abhishek Srivastava. "A Novel Hybrid Approach for Localization in Wireless Sensor Networks." In Communications in Computer and Information Science, 22–33. Cham: Springer Nature Switzerland, 2023. http://dx.doi.org/10.1007/978-3-031-25380-5_2.

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Yang, Gang, Shaohui Wu, Jieping Xu, and Xirong Li. "A Novel Hybrid SCH-ABC Approach for the Frequency Assignment Problem." In Neural Information Processing, 522–29. Berlin, Heidelberg: Springer Berlin Heidelberg, 2013. http://dx.doi.org/10.1007/978-3-642-42042-9_65.

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Conference papers on the topic "Novel hybrid approach"

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Dobrean, Dragoş, and Laura Dioşan. "A Hybrid Approach to MVC Architectural Layers Analysis." In 16th International Conference on Evaluation of Novel Approaches to Software Engineering. SCITEPRESS - Science and Technology Publications, 2021. http://dx.doi.org/10.5220/0010326700360046.

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Ranjzad, Hamed, Hossein Ebrahimnezhad, and Afshin Ebrahimi. "A Novel Compound Approach for Iris Segmentation." In 2008 Third International Conference on Convergence and Hybrid Information Technology (ICCIT). IEEE, 2008. http://dx.doi.org/10.1109/iccit.2008.176.

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Wang, Yongming, and Junzhong Gu. "A Novel Hybrid Approach for Diarrhea Prediction." In The 27th International Conference on Software Engineering and Knowledge Engineering. KSI Research Inc. and Knowledge Systems Institute Graduate School, 2015. http://dx.doi.org/10.18293/seke2015-102.

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Tiwari, Rajeev, and Neeraj Kumar. "A Novel Hybrid Approach for Web Caching." In 2012 Sixth International Conference on Innovative Mobile and Internet Services in Ubiquitous Computing (IMIS). IEEE, 2012. http://dx.doi.org/10.1109/imis.2012.39.

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Schäfer, Anna, Iurii Karpenko, and Hannah Elfner. "Conservation laws in a novel hybrid approach." In The International conference on Critical Point and Onset of Deconfinement. Trieste, Italy: Sissa Medialab, 2022. http://dx.doi.org/10.22323/1.400.0046.

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Dexheimer, Veronica, Stefan Schramm, and Jirina Stone. "A Novel Approach to Model Hybrid Stars." In XII International Symposium on Nuclei in the Cosmos. Trieste, Italy: Sissa Medialab, 2013. http://dx.doi.org/10.22323/1.146.0101.

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Aithal, Prakash K., G. Rajesh, P. C. Siddalingaswamy, and Dinesh U. Acharya. "A novel skew estimation approach using radon transform." In 2011 11th International Conference on Hybrid Intelligent Systems (HIS 2011). IEEE, 2011. http://dx.doi.org/10.1109/his.2011.6122070.

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Kahara, Timo, Keijo Haataja, and Pekka Toivanen. "A novel recommendation system approach utilizing social network profiles." In 2013 13th International Conference on Hybrid Intelligent Systems (HIS). IEEE, 2013. http://dx.doi.org/10.1109/his.2013.6920474.

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Hadj-Kacem, Mouna, and Nadia Bouassida. "A Hybrid Approach To Detect Code Smells using Deep Learning." In 13th International Conference on Evaluation of Novel Approaches to Software Engineering. SCITEPRESS - Science and Technology Publications, 2018. http://dx.doi.org/10.5220/0006709801370146.

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Liu, Yangxing, Takeshi Ikenaga, and Satoshi Goto. "A Novel Hybrid Approach of Color Image Segmentation." In APCCAS 2006 - 2006 IEEE Asia Pacific Conference on Circuits and Systems. IEEE, 2006. http://dx.doi.org/10.1109/apccas.2006.342202.

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Reports on the topic "Novel hybrid approach"

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Ostersetzer-Biran, Oren, and Jeffrey Mower. Novel strategies to induce male sterility and restore fertility in Brassicaceae crops. United States Department of Agriculture, January 2016. http://dx.doi.org/10.32747/2016.7604267.bard.

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Abstract Mitochondria are the site of respiration and numerous other metabolic processes required for plant growth and development. Increased demands for metabolic energy are observed during different stages in the plants life cycle, but are particularly ample during germination and reproductive organ development. These activities are dependent upon the tight regulation of the expression and accumulation of various organellar proteins. Plant mitochondria contain their own genomes (mtDNA), which encode for rRNAs, tRNAs and some mitochondrial proteins. Although all mitochondria have probably evolved from a common alpha-proteobacterial ancestor, notable genomic reorganizations have occurred in the mtDNAs of different eukaryotic lineages. Plant mtDNAs are notably larger and more variable in size (ranging from 70~11,000 kbp in size) than the mrDNAs in higher animals (16~19 kbp). Another unique feature of plant mitochondria includes the presence of both circular and linear DNA fragments, which undergo intra- and intermolecular recombination. DNA-seq data indicate that such recombination events result with diverged mitochondrial genome configurations, even within a single plant species. One common plant phenotype that emerges as a consequence of altered mtDNA configuration is cytoplasmic male sterility CMS (i.e. reduced production of functional pollen). The maternally-inherited male sterility phenotype is highly valuable agriculturally. CMS forces the production of F1 hybrids, particularly in predominantly self-pollinating crops, resulting in enhanced crop growth and productivity through heterosis (i.e. hybrid vigor or outbreeding enhancement). CMS lines have been implemented in some cereal and vegetables, but most crops still lack a CMS system. This work focuses on the analysis of the molecular basis of CMS. We also aim to induce nuclear or organellar induced male-sterility in plants, and to develop a novel approach for fertility restoration. Our work focuses on Brassicaceae, a large family of flowering plants that includes Arabidopsis thaliana, a key model organism in plant sciences, as well as many crops of major economic importance (e.g., broccoli, cauliflower, cabbage, and various seeds for oil production). In spite of the genomic rearrangements in the mtDNAs of plants, the number of genes and the coding sequences are conserved among different mtDNAs in angiosperms (i.e. ~60 genes encoding different tRNAs, rRNAs, ribosomal proteins and subunits of the respiratory system). Yet, in addition to the known genes, plant mtDNAs also harbor numerous ORFs, most of which are not conserved among species and are currently of unknown function. Remarkably, and relevant to our study, CMS in plants is primarily associated with the expression of novel chimericORFs, which likely derive from recombination events within the mtDNAs. Whereas the CMS loci are localized to the mtDNAs, the factors that restore fertility (Rfs) are identified as nuclear-encoded RNA-binding proteins. Interestingly, nearly all of the Rf’s are identified as pentatricopeptide repeat (PPR) proteins, a large family of modular RNA-binding proteins that mediate several aspects of gene expression primarily in plant organelles. In this project we proposed to develop a system to test the ability of mtORFs in plants, which are closely related to known CMS factors. We will induce male fertility in various species of Brassicaceae, and test whether a down-relation in the expression of the recombinantCMS-genes restores fertility, using synthetically designed PPR proteins.
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Gur, Amit, Edward Buckler, Joseph Burger, Yaakov Tadmor, and Iftach Klapp. Characterization of genetic variation and yield heterosis in Cucumis melo. United States Department of Agriculture, January 2016. http://dx.doi.org/10.32747/2016.7600047.bard.

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Project objectives: 1) Characterization of variation for yield heterosis in melon using Half-Diallele (HDA) design. 2) Development and implementation of image-based yield phenotyping in melon. 3) Characterization of genetic, epigenetic and transcriptional variation across 25 founder lines and selected hybrids. The epigentic part of this objective was modified during the course of the project: instead of characterization of chromatin structure in a single melon line through genome-wide mapping of nucleosomes using MNase-seq approach, we took advantage of rapid advancements in single-molecule sequencing and shifted the focus to Nanoporelong-read sequencing of all 25 founder lines. This analysis provides invaluable information on genome-wide structural variation across our diversity 4) Integrated analyses and development of prediction models Agricultural heterosis relates to hybrids that outperform their inbred parents for yield. First generation (F1) hybrids are produced in many crop species and it is estimated that heterosis increases yield by 15-30% globally. Melon (Cucumismelo) is an economically important species of The Cucurbitaceae family and is among the most important fleshy fruits for fresh consumption Worldwide. The major goal of this project was to explore the patterns and magnitude of yield heterosis in melon and link it to whole genome sequence variation. A core subset of 25 diverse lines was selected from the Newe-Yaar melon diversity panel for whole-genome re-sequencing (WGS) and test-crosses, to produce structured half-diallele design of 300 F1 hybrids (MelHDA25). Yield variation was measured in replicated yield trials at the whole-plant and at the rootstock levels (through a common-scion grafted experiments), across the F1s and parental lines. As part of this project we also developed an algorithmic pipeline for detection and yield estimation of melons from aerial-images, towards future implementation of such high throughput, cost-effective method for remote yield evaluation in open-field melons. We found extensive, highly heritable root-derived yield variation across the diallele population that was characterized by prominent best-parent heterosis (BPH), where hybrids rootstocks outperformed their parents by 38% and 56 % under optimal irrigation and drought- stress, respectively. Through integration of the genotypic data (~4,000,000 SNPs) and yield analyses we show that root-derived hybrids yield is independent of parental genetic distance. However, we mapped novel root-derived yield QTLs through genome-wide association (GWA) analysis and a multi-QTLs model explained more than 45% of the hybrids yield variation, providing a potential route for marker-assisted hybrid rootstock breeding. Four selected hybrid rootstocks are further studied under multiple scion varieties and their validated positive effect on yield performance is now leading to ongoing evaluation of their commercial potential. On the genomic level, this project resulted in 3 layers of data: 1) whole-genome short-read Illumina sequencing (30X) of the 25 founder lines provided us with 25 genome alignments and high-density melon HapMap that is already shown to be an effective resource for QTL annotation and candidate gene analysis in melon. 2) fast advancements in long-read single-molecule sequencing allowed us to shift focus towards this technology and generate ~50X Nanoporesequencing of the 25 founders which in combination with the short-read data now enable de novo assembly of the 25 genomes that will soon lead to construction of the first melon pan-genome. 3) Transcriptomic (3' RNA-Seq) analysis of several selected hybrids and their parents provide preliminary information on differentially expressed genes that can be further used to explain the root-derived yield variation. Taken together, this project expanded our view on yield heterosis in melon with novel specific insights on root-derived yield heterosis. To our knowledge, thus far this is the largest systematic genetic analysis of rootstock effects on yield heterosis in cucurbits or any other crop plant, and our results are now translated into potential breeding applications. The genomic resources that were developed as part of this project are putting melon in the forefront of genomic research and will continue to be useful tool for the cucurbits community in years to come.
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Hanson, Robert N. Solid Phase Combinatorial Approach to Estradiol Tamoxifen/Raloxifene Hybrids: Novel Chemotherapeutic/Prophylactic Selective Estrogen Receptor Modulators. Fort Belvoir, VA: Defense Technical Information Center, July 2002. http://dx.doi.org/10.21236/ada410893.

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Flaishman, Moshe, Herb Aldwinckle, Shulamit Manulis, and Mickael Malnoy. Efficient screening of antibacterial genes by juvenile phase free technology for developing resistance to fire blight in pear and apple trees. United States Department of Agriculture, December 2008. http://dx.doi.org/10.32747/2008.7613881.bard.

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Objectives: The original objectives of this project were to: Produce juvenile-free pear and apple plants and examine their sensitivity to E. amylovora; Design novel vectors, for antibacterial proteins and promoters expression, combined with the antisense TFL1 gene, and transformation of Spadona pear in Israel and Galaxy apple in USA. The original objectives were revised from the development of novel vectors with antibacterial proteins combined with the TFL-1 due to the inefficiency of alternative markes initially evaluated in pear, phoshomannose-isomerase and 2-deoxyglucose-6-phosphate phosphatase and the lack of development of double selection system. The objectives of project were revised to focus primarily on the development additional juvenile free systems by the use of another pear variety and manipulation of the FT gene under the control of several promoters. Based on the results creation of fire blight resistance pear variety was developed by the use of the juvenile free transgenic plant. Background: Young tree seedlings are unable to initiate reproductive organs and require a long period of shoot maturation, known as juvenile phase. In pear, juvenile period can last 5-7 years and it causes a major delay in breeding programs. We isolated the TFL1 gene from Spadona pear (PcTFL1-1) and produced transgenic ‘Spadona’ trees silencing the PcTFL1 gene using a RNAi approach. Transgenic tissue culture ‘Spadona’ pear flowered in vitro. As expected, the expression of the endogenous PcTFL1 was suppressed in the transgenic line that showed precocious flowering. Transgenic plants were successfully rooted in the greenhouse and most of the plants flowered after only 4-8 months, whereas the non-transformed control plants have flowered only after 5-6 years of development. Major achievements: Prior to flower induction, transgenic TFL1-RNAi ‘Spadona’ plants developed a few branches and leaves. Flower production in the small trees suppressed the development of the vegetative branches, thus resulting in compact flowering trees. Flowering was initiated in terminal buds, as described for the Arabidopsis tfl1 mutant. Propagation of the transgenic TFL1-RNAi ‘Spadona’ was performed by bud grafting on 'Betulifolia' rootstock and resulted in compact flowering trees. The transgenic flowering grafted plants were grown in the greenhouse under a long photoperiod for one year, and flowered continuously. Pollination of the transgenic flowers with ‘Costia‘ pear pollen generated fruits of regular shape with fertile F1 seeds. The F1 transgenic seedling grown in the greenhouse formed shoots and produced terminal flowers only five months after germination. In addition, grafted F1 transgenic buds flower and fruit continuously, generating hybrid fruits with regular shape, color and taste. Several pear varieties were pollinated with the transgenic TFL1-RNAi ‘Spadona’ pollen including `Herald Harw` that was reported to have resistance to fire blight diseases. The F-1 hybrid seedlings currently grow in our greenhouse. We conclude that the juvenile-free transgenic ‘Spadona’ pear enables the development of a fast breeding method in pear that will enable us to generate a resistance pear to fire blight. Implications: The research supported by this grant has demonstrated the use of transgenic juvenile free technology in pear. The use of the juvenile free technology for enhancement of conventional breeding in fruit tree will serve to enhance fast breeding systems in pear and another fruit trees.
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Avni, Adi, and Gitta L. Coaker. Proteomic investigation of a tomato receptor like protein recognizing fungal pathogens. United States Department of Agriculture, January 2015. http://dx.doi.org/10.32747/2015.7600030.bard.

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Maximizing food production with minimal negative effects on the environment remains a long-term challenge for sustainable food production. Microbial pathogens cause devastating diseases, minimizing crop losses by controlling plant diseases can contribute significantly to this goal. All plants possess an innate immune system that is activated after recognition of microbial-derived molecules. The fungal protein Eix induces defense responses in tomato and tobacco. Plants recognize Eix through a leucine-rich-repeat receptor- like-protein (LRR-RLP) termed LeEix. Despite the knowledge obtained from studies on tomato, relatively little is known about signaling initiated by RLP-type immune receptors. The focus of this grant proposal is to generate a foundational understanding of how the tomato xylanase receptor LeEix2 signals to confer defense responses. LeEix2 recognition results in pattern triggered immunity (PTI). The grant has two main aims: (1) Isolate the LeEix2 protein complex in an active and resting state; (2) Examine the biological function of the identified proteins in relation to LeEix2 signaling upon perception of the xylanase elicitor Eix. We used two separate approaches to isolate receptor interacting proteins. Transgenic tomato plants expressing LeEix2 fused to the GFP tag were used to identify complex components at a resting and activated state. LeEix2 complexes were purified by mass spectrometry and associated proteins identified by mass spectrometry. We identified novel proteins that interact with LeEix receptor by proteomics analysis. We identified two dynamin related proteins (DRPs), a coiled coil – nucleotide binding site leucine rich repeat (SlNRC4a) protein. In the second approach we used the split ubiquitin yeast two hybrid (Y2H) screen system to identified receptor-like protein kinase At5g24010-like (SlRLK-like) (Solyc01g094920.2.1) as an interactor of LeEIX2. We examined the role of SlNRC4a in plant immunity. Co-immunoprecipitation demonstrates that SlNRC4a is able to associate with different PRRs. Physiological assays with specific elicitors revealed that SlNRC4a generally alters PRR-mediated responses. SlNRC4a overexpression enhances defense responses while silencing SlNRC4 reduces plant immunity. We propose that SlNRC4a acts as a non-canonical positive regulator of immunity mediated by diverse PRRs. Thus, SlNRC4a could link both intracellular and extracellular immune perception. SlDRP2A localizes at the plasma membrane. Overexpression of SlDRP2A increases the sub-population of LeEIX2 inVHAa1 endosomes, and enhances LeEIX2- and FLS2-mediated defense. The effect of SlDRP2A on induction of plant immunity highlights the importance of endomembrane components and endocytosis in signal propagation during plant immune . The interaction of LeEIX2 with SlRLK-like was verified using co- immunoprecipitation and a bimolecular fluorescence complementation assay. The defence responses induced by EIX were markedly reduced when SlRLK-like was over-expressed, and mutation of slrlk-likeusing CRISPR/Cas9 increased EIX- induced ethylene production and SlACSgene expression in tomato. Co-expression of SlRLK-like with different RLPs and RLKs led to their degradation, apparently through an endoplasmic reticulum-associated degradation process. We provided new knowledge and expertise relevant to expression of specific be exploited to enhance immunity in crops enabling the development of novel environmentally friendly disease control strategies.
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Fridman, Eyal, Jianming Yu, and Rivka Elbaum. Combining diversity within Sorghum bicolor for genomic and fine mapping of intra-allelic interactions underlying heterosis. United States Department of Agriculture, January 2012. http://dx.doi.org/10.32747/2012.7597925.bard.

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Heterosis, the enigmatic phenomenon in which whole genome heterozygous hybrids demonstrate superior fitness compared to their homozygous parents, is the main cornerstone of modern crop plant breeding. One explanation for this non-additive inheritance of hybrids is interaction of alleles within the same locus. This proposal aims at screening, identifying and investigating heterosis trait loci (HTL) for different yield traits by implementing a novel integrated mapping approach in Sorghum bicolor as a model for other crop plants. Originally, the general goal of this research was to perform a genetic dissection of heterosis in a diallel built from a set of Sorghum bicolor inbred lines. This was conducted by implementing a novel computational algorithm which aims at associating between specific heterozygosity found among hybrids with heterotic variation for different agronomic traits. The initial goals of the research are: (i) Perform genotype by sequencing (GBS) of the founder lines (ii) To evaluate the heterotic variation found in the diallel by performing field trails and measurements in the field (iii) To perform QTL analysis for identifying heterotic trait loci (HTL) (iv) to validate candidate HTL by testing the quantitative mode of inheritance in F2 populations, and (v) To identify candidate HTL in NAM founder lines and fine map these loci by test-cross selected RIL derived from these founders. The genetic mapping was initially achieved with app. 100 SSR markers, and later the founder lines were genotyped by sequencing. In addition to the original proposed research we have added two additional populations that were utilized to further develop the HTL mapping approach; (1) A diallel of budding yeast (Saccharomyces cerevisiae) that was tested for heterosis of doubling time, and (2) a recombinant inbred line population of Sorghum bicolor that allowed testing in the field and in more depth the contribution of heterosis to plant height, as well as to achieve novel simulation for predicting dominant and additive effects in tightly linked loci on pseudooverdominance. There are several conclusions relevant to crop plants in general and to sorghum breeding and biology in particular: (i) heterosis for reproductive (1), vegetative (2) and metabolic phenotypes is predominantly achieved via dominance complementation. (ii) most loci that seems to be inherited as overdominant are in fact achieving superior phenotype of the heterozygous due to linkage in repulsion, namely by pseudooverdominant mechanism. Our computer simulations show that such repulsion linkage could influence QTL detection and estimation of effect in segregating populations. (iii) A new height QTL (qHT7.1) was identified near the genomic region harboring the known auxin transporter Dw3 in sorghum, and its genetic dissection in RIL population demonstrated that it affects both the upper and lower parts of the plant, whereas Dw3 affects only the part below the flag leaf. (iv) HTL mapping for grain nitrogen content in sorghum grains has identified several candidate genes that regulate this trait, including several putative nitrate transporters and a transcription factor belonging to the no-apical meristem (NAC)-like large gene family. This activity was combined with another BARD-funded project in which several de-novo mutants in this gene were identified for functional analysis.
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Perdigão, Rui A. P. Earth System Dynamic Intelligence - ESDI. Meteoceanics, April 2021. http://dx.doi.org/10.46337/esdi.210414.

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Earth System Dynamic Intelligence (ESDI) entails developing and making innovative use of emerging concepts and pathways in mathematical geophysics, Earth System Dynamics, and information technologies to sense, monitor, harness, analyze, model and fundamentally unveil dynamic understanding across the natural, social and technical geosciences, including the associated manifold multiscale multidomain processes, interactions and complexity, along with the associated predictability and uncertainty dynamics. The ESDI Flagship initiative ignites the development, discussion and cross-fertilization of novel theoretical insights, methodological developments and geophysical applications across interdisciplinary mathematical, geophysical and information technological approaches towards a cross-cutting, mathematically sound, physically consistent, socially conscious and operationally effective Earth System Dynamic Intelligence. Going beyond the well established stochastic-dynamic, information-theoretic, artificial intelligence, mechanistic and hybrid techniques, ESDI paves the way to exploratory and disruptive developments along emerging information physical intelligence pathways, and bridges fundamental and operational complex problem solving across frontier natural, social and technical geosciences. Overall, the ESDI Flagship breeds a nascent field and community where methodological ingenuity and natural process understanding come together to shed light onto fundamental theoretical aspects to build innovative methodologies, products and services to tackle real-world challenges facing our planet.
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Dolja, Valerian V., Amit Gal-On, and Victor Gaba. Suppression of Potyvirus Infection by a Closterovirus Protein. United States Department of Agriculture, March 2002. http://dx.doi.org/10.32747/2002.7580682.bard.

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The plant virus family Polyviridae is the largest and most destructive of all plant viruses. Despite the continuous effort to develop resistant plant varieties, there is a desperate need for novel approaches conferring wide-range potyvirus resistance. Based on experiments with the tobacco etch potyvirus (TEV)-derived gene expression vector, we suggested approach for screening of the candidate resistance genes. This approach relies on insertion of the genes into a virus vector and evaluation of the phenotypes of the resulting recombinant viruses. The genes which suppress infection by the recombinant virus are selected as candidates for engineering transgenic resistance. Our analysis of the TEV variants expressing proteins of the beet yellows closterovirus (BYV) revealed that one of those, the leader proteinase (L-Pro), strongly and specifically interfered with the hybrid TEV infection. Since closterovirus L-Pro is evolutionary related to potyviral helper component-proteinase (HC-Pro), we suggested that the L-Pro interfered with HC-Pro function via a trans-dominant inhibitory effect. Based on these findings, we proposed to test two major hypotheses. First, we suggested that L-Pro-mediated suppression of potyvirus infection is a general phenomenon effective against a range of potyviruses. The second hypothesis stated that the suppression effect can be reproduced in transgenic plants expressing L-Pro, and can be utilized for generation of resistance to potyviruses. In accord with these hypotheses, we developed two original objectives of our proposal: A) to determine the range of the closterovirus-derived suppression of potyviral infection, and B) to try and utilize the L-Pro-mediated suppression for the development of transgenic resistance to potyviruses. In the first phase of the project, we have developed all major tools and technologies required for successful completion of the proposed research. These included TEV and ZYMV vectors engineered to express several closteroviral L-Pro variants, and generation of the large collection of transgenic plants. To our satisfaction, characterization of the infection phenotypes exhibited by chimeric TEV and ZYMV variants confirmed our first hypothesis. For instance, similar to TEV-L- Pro(BYV) chimera, ZYMV-L-Pro(LIYV) chimera was debilitated in its systemic spread. In contrast, ZYMV-GUS chimera (positive control) was competent in establishing vigorous systemic infection. These and other results with chimeric viruses indicated that several closteroviral proteinases inhibit long-distance movement of the potyviruses upon co-expression in infected plants. In order to complete the second objective, we have generated ~90 tobacco lines transformed with closteroviral L-Pro variants, as well as ~100 lines transformed with BYV Hsp70-homolog (Hsp70h; a negative control). The presence and expression of the trans gene in each line was initially confirmed using RT-PCR and RNA preparations isolated from plants. However, since detection of the trans gene-specific RNA can not guarantee production of the corresponding protein, we have also generated L-Pro- and Hsp70h-specific antisera using corresponding synthetic peptides. These antisera allowed us to confirm that the transgenic plant lines produced detectable, although highly variable levels of the closterovirus antigens. In a final phase of the project, we tested susceptibility of the transgenic lines to TEV infection. To this end, we determined that the minimal dilution of the TEV inoculum that is still capable of infecting 100% of nontransgenic plants was 1:20, and used 10 plants per line (in total, ~2,000 plants). Unfortunately, none of the lines exhibited statistically significant reduction in susceptibility. Although discouraging, this outcome prompted us to expand our experimental plan and conduct additional experiments. Our aim was to test if closteroviral proteinases are capable of functioning in trans. We have developed agroinfection protocol for BYV, and tested if co- expression of the L-Pro is capable of rescuing corresponding null-mutant. The clear-cut, negative results of these experiments demonstrated that L-Pro acts only in cis, thus explaining the lack of resistance in our transgenic plants. We have also characterized a collection of the L-Pro alanine- scanning mutants and found direct genetic evidence of the requirement for L-Pro in virus systemic spread. To conclude, our research supported by BARD confirmed one but not another of our original hypotheses. Moreover, it provided an important insight into functional specialization of the viral proteinases and generated set of tools and data with which we will be able to address the molecular mechanisms by which these proteins provide a variety of critical functions during virus life cycle.
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Ohad, Nir, and Robert Fischer. Regulation of Fertilization-Independent Endosperm Development by Polycomb Proteins. United States Department of Agriculture, January 2004. http://dx.doi.org/10.32747/2004.7695869.bard.

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Arabidopsis mutants that we have isolated, encode for fertilization-independent endosperm (fie), fertilization-independent seed2 (fis2) and medea (mea) genes, act in the female gametophyte and allow endosperm to develop without fertilization when mutated. We cloned the FIE and MEA genes and showed that they encode WD and SET domain polycomb (Pc G) proteins, respectively. Homologous proteins of FIE and MEA in other organisms are known to regulate gene transcription by modulating chromatin structure. Based on our results, we proposed a model whereby both FIE and MEA interact to suppress transcription of regulatory genes. These genes are transcribed only at proper developmental stages, as in the central cell of the female gametophyte after fertilization, thus activating endosperm development. To test our model, the following questions were addressed: What is the Composition and Function of the Polycomb Complex? Molecular, biochemical, genetic and genomic approaches were offered to identify members of the complex, analyze their interactions, and understand their function. What is the Temporal and Spatial Pattern of Polycomb Proteins Accumulation? The use of transgenic plants expressing tagged FIE and MEA polypeptides as well as specific antibodies were proposed to localize the endogenous polycomb complex. How is Polycomb Protein Activity Controlled? To understand the molecular mechanism controlling the accumulation of FIE protein, transgenic plants as well as molecular approaches were proposed to determine whether FIE is regulated at the translational or posttranslational levels. The objectives of our research program have been accomplished and the results obtained exceeded our expectation. Our results reveal that fie and mea mutations cause parent-of-origin effects on seed development by distinct mechanisms (Publication 1). Moreover our data show that FIE has additional functions besides controlling the development of the female gametophyte. Using transgenic lines in which FIE was not expressed or the protein level was reduced during different developmental stages enabled us for the first time to explore FIE function during sporophyte development (Publication 2 and 3). Our results are consistent with the hypothesis that FIE, a single copy gene in the Arabidopsis genome, represses multiple developmental pathways (i.e., endosperm, embryogenesis, shot formation and flowering). Furthermore, we identified FIE target genes, including key transcription factors known to promote flowering (AG and LFY) as well as shoot and leaf formation (KNAT1) (Publication 2 and 3), thus demonstrating that in plants, as in mammals and insects, PcG proteins control expression of homeobox genes. Using the Yeast two hybrid system and pull-down assays we demonstrated that FIE protein interact with MEA via the N-terminal region (Publication 1). Moreover, CURLY LEAF protein, an additional member of the SET domain family interacts with FIE as well. The overlapping expression patterns of FIE, with ether MEA or CLF and their common mutant phenotypes, demonstrate the versatility of FIE function. FIE association with different SET domain polycomb proteins, results in differential regulation of gene expression throughout the plant life cycle (Publication 3). In vitro interaction assays we have recently performed demonstrated that FIE interacts with the cell cycle regulatory component Retinobalsoma protein (pRb) (Publication 4). These results illuminate the potential mechanism by which FIE may restrain embryo sac central cell division, at least partly, through interaction with, and suppression of pRb-regulated genes. The results of this program generated new information about the initiation of reproductive development and expanded our understanding of how PcG proteins regulate developmental programs along the plant life cycle. The tools and information obtained in this program will lead to novel strategies which will allow to mange crop plants and to increase crop production.
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Coplin, David L., Shulamit Manulis, and Isaac Barash. roles Hrp-dependent effector proteins and hrp gene regulation as determinants of virulence and host-specificity in Erwinia stewartii and E. herbicola pvs. gypsophilae and betae. United States Department of Agriculture, June 2005. http://dx.doi.org/10.32747/2005.7587216.bard.

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Gram-negative plant pathogenic bacteria employ specialized type-III secretion systems (TTSS) to deliver an arsenal of pathogenicity proteins directly into host cells. These secretion systems are encoded by hrp genes (for hypersensitive response and pathogenicity) and the effector proteins by so-called dsp or avr genes. The functions of effectors are to enable bacterial multiplication by damaging host cells and/or by blocking host defenses. We characterized essential hrp gene clusters in the Stewart's Wilt of maize pathogen, Pantoea stewartii subsp. stewartii (Pnss; formerly Erwinia stewartii) and the gall-forming bacterium, Pantoea agglomerans (formerly Erwinia herbicola) pvs. gypsophilae (Pag) and betae (Pab). We proposed that the virulence and host specificity of these pathogens is a function of a) the perception of specific host signals resulting in bacterial hrp gene expression and b) the action of specialized signal proteins (i.e. Hrp effectors) delivered into the plant cell. The specific objectives of the proposal were: 1) How is the expression of the hrp and effector genes regulated in response to host cell contact and the apoplastic environment? 2) What additional effector proteins are involved in pathogenicity? 3) Do the presently known Pantoea effector proteins enter host cells? 4) What host proteins interact with these effectors? We characterized the components of the hrp regulatory cascade (HrpXY ->7 HrpS ->7 HrpL ->7 hrp promoters), showed that they are conserved in both Pnss and Fag, and discovered that the regulation of the hrpS promoter (hrpSp) may be a key point in integrating apoplastic signals. We also analyzed the promoters recognized by HrpL and demonstrated the relationship between their composition and efficiency. Moreover, we showed that promoter strength can influence disease expression. In Pnss, we found that the HrpXY two-component signal system may sense the metabolic status of the bacterium and is required for full hrp gene expression in planta. In both species, acyl-homoserine lactone-mediated quorum sensing may also regulate epiphytic fitness and/or pathogenicity. A common Hrp effector protein, DspE/WtsE, is conserved and required for virulence of both species. When introduced into corn cells, Pnss WtsE protein caused water-soaked lesions. In other plants, it either caused cell death or acted as an Avr determinant. Using a yeast- two-hybrid system, WtsE was shown to interact with a number of maize signal transduction proteins that are likely to have roles in either programmed cell death or disease resistance. In Pag and Pab, we have characterized the effector proteins HsvG, HsvB and PthG. HsvG and HsvB are homologous proteins that determine host specificity of Pag and Pab on gypsophila and beet, respectively. Both possess a transcriptional activation domain that functions in yeast. PthG was found to act as an Avr determinant on multiple beet species, but was required for virulence on gypsophila. In addition, we demonstrated that PthG acts within the host cell. Additional effector genes have been characterized on the pathogenicity plasmid, pPATHₚₐg, in Pag. A screen for HrpL- regulated genes in Pnsspointed up 18 candidate effector proteins and four of these were required for full virulence. It is now well established that the virulence of Gram-negative plant pathogenic bacteria is governed by Hrp-dependent effector proteins. However; the mode of action of many effectors is still unresolved. This BARD supported research will significantly contribute to the understanding of how Hrp effectors operate in Pantoea spp. and how they control host specificity and affect symptom production. This may lead to novel approaches for genetically engineering plants resistant to a wide range of bacterial pathogens by inactivating the Hrp effectors with "plantabodies" or modifying their receptors, thereby blocking the induction of the susceptible response. Alternatively, innovative technologies could be used to interfere with the Hrp regulatory cascade by blocking a critical step or mimicking plant or quorum sensing signals.
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