{"pk":41374,"title":"Genetic transformation of sweet orange to overexpress a CsPR-8 gene aiming for Candidatus Liberibacter asiaticus resistance","subtitle":null,"abstract":"A strategy to produce HLB-resistant citrus using genetic engineering is the overexpression of genes identified in the citrus genome. Plants respond to pathogen attacks by producing several pathogenesis-related (PR) proteins. Therefore, individual PR overexpression in transgenic plants can lead to an increased resistance. In this study, we have chosen to use one \nPR-8\n isoform cloned from \nCitrus sinensis \n(\nCsPR-8\n). The PR-8 is an endochitinase that also has lysozyme activity, to be potentially used against bacterial attacks. We constructed an expression transformation vector (pCAMBIA2201) containing the \nCsPR-8\n gene and the selection gene \nnpt\nII that confers kanamycin resistance in plants, both driven by the CaMV35S constitutive promoter. Epicotyl segments collected from \nin vitro \nseedlings of ‘Hamlin’ sweet orange (\nCitrus sinensis\n L. Osbeck) were used for transformation via \nAgrobacterium tumefaciens\n strain EHA105. The developed shoots were excised from the explants and \nin vitro\n grafted onto Carrizo citrange [\nC. sinensis\n x \nPoncirus trifoliata\n (L.) Raf] seedlings. The grafted plants were analyzed by PCR, using specific primers for detection of the \nnpt\nII gene. Acclimation of transgenic plants is under way in order to be transferred to the greenhouse. These plants will be analyzed by Southern blot to confirm the integration of the transgene and by RT-qPCR to evaluate the transgene expression, prior to their evaluation for \nCandidatus\n Liberibacter asiaticus resistance.","language":"en","license":{"name":"Creative Commons Attribution 4.0","short_name":"CC BY 4.0","text":"Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.\n\nNo additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.","url":"https://creativecommons.org/licenses/by/4.0"},"keywords":[],"section":"Abstracts of Presentations at the 3rd International Research Conference on Huanglongbing","is_remote":true,"remote_url":"https://escholarship.org/uc/item/9bz1w1hc","frozenauthors":[{"first_name":"F.","middle_name":"A.A.","last_name":"Mourão Filho","name_suffix":"","institution":"Universidade de São Paulo/ESALQ, Piracicaba, Brazil","department":"None"},{"first_name":"L.","middle_name":"C.L.","last_name":"Stipp","name_suffix":"","institution":"Universidade de São Paulo/ESALQ, Piracicaba, Brazil","department":"None"},{"first_name":"A.","middle_name":"B.","last_name":"Beltrame","name_suffix":"","institution":"Universidade de São Paulo/ESALQ, Piracicaba, Brazil","department":"None"},{"first_name":"R.","middle_name":"L.","last_name":"Boscariol-Camargo","name_suffix":"","institution":"Centro de Citricultura Sylvio Moreira/IAC, Cordeirópolis, Brazil","department":"None"},{"first_name":"R.","middle_name":"","last_name":"Harakava","name_suffix":"","institution":"Instituto Biológico, Seção de Bioquímica Fitopatológica, São Paulo, Brazil","department":"None"},{"first_name":"B.","middle_name":"M.J.","last_name":"Mendes","name_suffix":"","institution":"Universidade de São Paulo/CENA, Piracicaba, Brazil","department":"None"}],"date_submitted":"2014-12-23T00:08:02Z","date_accepted":"2014-12-23T00:08:02Z","date_published":"2014-01-01T00:00:00Z","render_galley":null,"galleys":[{"label":"","type":"pdf","path":"https://journalpub.escholarship.org/iocv_journalcitruspathology/article/41374/galley/30973/download/"}]}