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The Study of Fasciation

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The Study of Fasciation Effectiveness of Selection Methods for Improvement of Portuguese Maize (Zea mays L.) The study of fasciation Pedro Manuel Reis Mendes Moreira Dissertation presented to obtain the Ph.D degree in Biology (Population Genetics) Instituto de Tecnologia Química e Biológica António Xavier | Universidade Nova de Lisboa Oeiras, “July 2015” PhD Supervisors: Dr. Maria Carlota Vaz Patto (Supervisor) Prof. Pedro Fevereiro (Co-Supervisor) The work presented in this thesis was performed at: Instituto de Tecnologia Química e Biológica António Xavier (ITQB) Instituto Politécnico de Coimbra - Escola Superior Agrária de Coimbra PhD thesis Advisory Board Prof. Arnel R.Hallauer Dr. Silas E. Pêgo VASO PPB Project Since 1984 ii The student, Pedro Manuel Reis Mendes Moreira received financial support from Fundação para a Ciência e Tecnologia and Fundo Social Europeu in the scope of Quadro Comunitário de Apoio through the PhD fellowship SFRH/BD/22188/2005 and in the scope of programa de apoio à formação avançada de docentes do Ensino Superior Politécnico SFRH/PROTEC/50365/2009 Additional Funding: FCT project « Analysis of the breeding potential of fasciation traits in Portuguese maize landraces » (PTDC/AGR-AAM/70845/2006) (2007-2010) Chapter III presentation was sponsored by Cost Action 860 Short-term grant by Zagreb University, under guidance of Zlatko Šatović ERASMUS Program OpenDipSA - ERASMUS 22-23 July 2013 University of Bologna European Commission through the FP7-KBBE- 2009-3, under the grant agreement no. 245058, SOLIBAM project iii “Advances in medicine and agriculture have saved vastly more lives than have been lost in all the wars in history.” ― Carl Sagan, The Demon-Haunted World: Science as a Candle in the Dark iv v To Daniela, Lucas, Sara and Francisca vi Acknowledgments After my stage at NUMI/BPGV since 1995 to 1997 under the guidance of Silas Pêgo, many attempts were made to start the PhD program. In 29 March 2003 the start of this journey was initiated with the submission of a poster to Arnel R. Hallauer International Symposium on Plant Breeding held at the Hotel Camino Real in Mexico City from 17-22 August, 2003. At this Symposium I had the opportunity to meet Arnel R. Hallauer, which resulted in two actions, my stage in 2004 at Iowa State University at Kendall Lamkey maize breeding program with a FLAD support and in the invitation to Arnel R. Hallauer to come to Portugal that was held at same year. During this time I had the opportunity to invite Arnel R. Hallauer as PhD. Advisor together with Silas E. Pêgo. In parallel the project POCTI/AGR/57994/2004 “Portuguese maize bread quality: from technological aspects to controlling genes”, had started in 1st of February of 2005 with Maria Carlota Vaz Patto as Project Coordinator. Due to Maria Carlota Vaz Patto expertise and interest on the maize breeding topic she was the skilled and right person as PhD Supervisor together with Pedro Fevereiro as PhD Co-supervisor on the molecular analysis that were missing on the fasciation trait. This was also the reason that we apply to both applications to FCT, both succeeded, throughout PhD fellowship and later by the project «Analysis of the breeding potential of fasciation traits in Portuguese maize landraces» vii (PTDC/AGR-AAM/70845/2006) coordinated also by MC Vaz Patto, that were crucial for the development of this thesis. This story gives the context to acknowledge to my Supervisors and Advisors for their acceptance and valuable contributions, which were crucial in each step of PhD work. Without them this work would not be possible. - Silas Pêgo, for his constant help motivation and availability, guidance, valuable corrections and suggestions and wisdom for starting the VASO PPB Project, one of the oldest in the world. - Arnel Hallauer, for all the support, guidance and valuable corrections and suggestions of the manuscripts. - Pedro Salema Fevereiro, for his acceptance and support in the PhD work. - Carlota Vaz Patto for her constant help, support and guidance, since the lab work interpretation of results and a crucial help on the paper submissions and thesis elaboration. My experience at Iowa State University with Arnel Hallauer is naturally extended to Kendal Lamkey and team from Iowa State University that kindly accepted me in his program and later he kindly accepted to perform part of multilocation trials in USA during 2005, described in Chapter III and Chapter IV. To BCV, ITQB personal (Nuno Almeida, Mara Alves, Rita Caré) for their untiring tutoring, support and teachings in the lab, constant encouragement and meaningful advice before and after the lab. To Zlatko Šatović, for his kind support, and fruitfull discutions at Zagreb University via his guidance and scholarship that made possible and very pleasant my stage at his lab, 谢谢. viii To João Pedro Mendes Moreira for the presential and tele meetings that were very important to create a common language of discussion especially important for Chapter VI. To my students and collaborators: João Santos, J.P. Santos, Marisa Antunes, Liliana Morgado, Ralf Moura, Cláudia Brites, Manuel Paulo, Daniel Gaspar, Graciano Spencer, Joana Laires … To collaborators Engº Vaz Patto, Engª Rosa Guilherme from IPC-ESAC. In memoriam to Wayne Haag (as CIMMYT director of maize breeding in the Mediterranean area in 1985) who first supported the VASO project. To the farmers and specially Mr. Francisco Meireles for its critical analysis and empirical knowledge, that made VASO project a bridge between empirical knowledge and science… and probably a “seed” for Socionomics in Participatory Plant Breeding. To my friends that do not see me for a while … even the nearest ones… but that were encouraging and supporting me all the time. To my parents for their solid contribution to my G, E and G x E. To my father for his constant learning encouragement and perseverance. To Daniela, Lucas, Sara and Francisca that also started the PhD with me… A Todos o meu Muito Obrigado. ix Sumário Desde a sua introdução, pós Colombo, o milho provocou uma revolução silenciosa sobre a região Norte e Centro de Portugal. Revolução que se traduziu pela reformulação dos sistemas de cultivo, agronomia, paisagem e cultura ao longo dos anos. Na década de 1940, o sucesso das sementes híbridas americanas iniciou o seu contributo para a erosão genética. No NUMI (Estação de melhoramento de milho em Braga) Silas Pêgo compreendeu desde logo esta ameaça. Assim, várias missões de colheita de germoplasma de milho foram organizadas. Esta recolha contribuiu para a conservação ex-situ; tendo contribuído também para as atividades in situ / on-farm e on-station via pré-melhoramento. O presente trabalho inicia-se com a descrição do projeto VASO (um projeto de Melhoramento Participativo de Plantas), iniciado em 1984 na região do Vale do Sousa onde a conservação da diversidade genética e atividades de melhoramento continuam, tendo por objetivo a qualidade do milho para "broa" (pão de milho). O projeto VASO representa também uma oportunidade para a adaptação do germoplasma às áreas marginais de produção, agricultura sustentável e integração do conhecimento tradicional. O nosso trabalho prosseguiu com uma caracterização detalhada do trabalho de melhoramento participativo do projeto VASO desde a sua génese. A abordagem quantitativa seguida, permitiu comparar os x métodos de seleção aplicados pelo melhorador e agricultor, utilizando 'Pigarro’ "(variedade regional portuguesa de milho liso, branco) e ‘Fandango’ (uma população sintética de milho amarelo dentado). Como resultado, os agricultores selecionaram espigas mais curtas e largas, com aumento dos níveis de fasciação e grãos de menor dimensão. No caso da seleção do melhorador, as espigas tornaram-se mais compridas e menos fasciadas, com um aumento da uniformidade da cultura. Ambos os métodos de seleção do melhorador e agricultor foram eficazes para a conservação da diversidade. Deste modo, a escolha do método de seleção, dependerá dos objetivos do programa de melhoramento: seleção fenotípica recorrente é mais fácil e potencialmente mais económica para adotar pelos agricultores para melhoramento de OPVs (variedades de polinização livre), enquanto que os resultados de seleção pelo melhorador resultam numa maior uniformidade da cultura, estando mais adaptadas para programas de desenvolvimento de híbridos. A análise da evolução da diversidade molecular enfatizou associações potenciais entre determinados marcadores moleculares neutros e os loci responsáveis pelo controlo de algumas das características fenotípicas sob seleção (e.g., comprimento da espiga, fasciação e características associadas à espiga como o diâmetro da espiga e número de grãos por carreira). Estas associações precisam no entanto de ser melhor analisadas e validadas através de xi mapeamento de ligação ou associação, de modo a ancorarem a seleção de características a sistemas de agricultura sustentáveis. O nosso trabalho permitiu ainda que os dados fenotípicos fossem utilizados no desenvolvimento de ferramentas de seleção para os agricultores, contribuindo para melhorar o processo de seleção. Com este propósito, melhoramos a "fórmula do valor da espiga", como ferramenta de seleção do agricultor, procurando aumentar a produção com base nas características da espiga. O valor da Espiga (EV) foi desenvolvido em 1993 no âmbito de um concurso regional de espigas de milho (Concurso da “Melhor Espiga do Vale do Sousa”). Esta fórmula tinha dois objetivos principais, a avaliação de espigas para o referido concurso e como ferramenta pedagógica no melhoramento de milho por parte dos agricultores. A fórmula EV foi baseada em correlações de características de milho publicadas na literatura, sem que constassem dados de campo como a produção. Para cumprir esta lacuna geramos métodos diferentes e desenvolvemos um método de classificação para compará-los. Utilizamos para tal os dados de um conjunto de populações de onde as melhores espigas do Vale do Sousa provieram. A partir dos métodos utilizados, a fórmula EVA foi a escolhida por ser facilmente adotada pelos agricultores e associações interessadas na conservação e no desenvolvimento de germoplasma.
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