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ANATOMICAL and PHYSIOLOGICAL RESPONSES of Billbergia Zebrina (Bromeliaceae) UNDER DIFFERENT in VITRO CONDITIONS

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JOÃO PAULO RODRIGUES MARTINS ANATOMICAL AND PHYSIOLOGICAL RESPONSES OF Billbergia zebrina (Bromeliaceae) UNDER DIFFERENT IN VITRO CONDITIONS LAVRAS- MG 2015 JOÃO PAULO RODRIGUES MARTINS ANATOMICAL AND PHYSIOLOGICAL RESPONSES OF Billbergia zebrina (BROMELIACEAE) UNDER DIFFERENT IN VITRO CONDITIONS This thesis is being submitted in a partial fulfilment of the requirements for degree of Doctor in Applied Botanic of Universidade Federal de Lavras. Supervisor Dr. Moacir Pasqual Co-supervisor Dr. Maurice De Proft LAVRAS- MG 2015 Ficha catalográfica elaborada pelo Sistema de Geração de Ficha Catalográfica da Biblioteca Universitária da UFLA, com dados informados pelo(a) próprio(a) autor(a). Martins, João Paulo Rodrigues. Anatomical and physiological responses of Billbergia zebrina (Bromeliaceae) under different in vitro conditions / João Paulo Rodrigues Martins. – Lavras : UFLA, 2015. 136 p. : il. Tese(doutorado)–Universidade Federal de Lavras, 2015. Orientador(a): Moacir Pasqual. Bibliografia. 1. Bromeliad. 2. In vitro culture. 3. Photoautotrophic growth. 4. Plant anatomy. 5. Plant physiology. I. Universidade Federal de Lavras. II. Título. JOÃO PAULO RODRIGUES MARTINS ANATOMICAL AND PHYSIOLOGICAL RESPONSES OF Billbergia zebrina (BROMELIACEAE) UNDER DIFFERENT IN VITRO CONDITIONS This thesis is being submitted in a partial fulfilment of the requirements for degree of Doctor in Applied Botanic of Universidade Federal de Lavras. APPROVED 09th of June, 2015 Dr Diogo Pedrosa Corrêa da Silva UFLA Dra Leila Aparecida Salles Pio UFLA Dr Thiago Corrêa de Souza UNIFAL-MG Dra Vânia Helena Techio UFLA Dra Cynthia de Oliveira UFLA Supervisor Dr. Moacir Pasqual Co-supervisor Dr. Maurice De Proft LAVRAS- MG 2015 ACKNOWLEDGEMENTS God for having guided my path. My wonderful family (Including Capivara), I could not ask for better people. Thanks for supporting me and I apologise for my absence during my academic life. My grandmother Maria Ana (in memoriam). She pushed me to study and she will always be the most important person in my life. Thanks! CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior) for granting me a scholarship. Eliana and Fernando, I will always be thankful for your help during my PhD. Italo, Claret and Vantuil, thanks you all support in the lab, I could not finish my thesis without your help. My promoter in Brazil, Dr. Moacir Pasqual, for all confidence in the development of the thesis project. My colleagues of NECULT, especially Adalvan, for all help in our lab. Thanks for that! All master and PhD students of Applied Botanic Post-graduation Program. You are great friends and I will never forget the farewell party you have done to me. I hope to keep our friendship for a very long time. Thanks for sharing your time with me. My most sincere thanks to Alexandra Nunes, Carol, Cíntia, Diego, Dorfo, Flávia, Gabriel, Jaiane, Jaque, Jean, Katixa, Laiane, Larissa, Luiz, Mairy, Marcio, Marinês, Mayara, Nayara, Suelen, and Thais. I could not spend my time at many bars with better people. A special thanks to Marinês, who pushed me to PhD, I would not be here without her. Pedro, you are a loyal friend, thanks for your help and time at Meu Cantinho and other places. I really appreciate it. Martin and his family, they gave me one of the nicest Christmas of my life. Thanks Martin, you pushed me to Europe, I will always be in debt to you for it. My friends from Alfenas, Kellen, Lucas, Marina, Pitol, Samara and Vanessa. We have been friends for years and part of this thesis came from you. You are my inspiration to keep trying to be a better person each day. Maurice, you were more than a promoter during my PhD sandwich in Belgium, you were a father. And if I had a father, I cannot complain, I had a Belgian mother as well. Thanks Veerle! I had my sister and brothers too, Ruth, Willem and Bart. Thanks you all for being so kind to me. I also thank my good friends from Europe. Ana, Anže, Bart Panis, Bie, Carlo, Clara, Deby, Delfina, Dieter, Ewaut, Francesco, Jassmine, Jelena, Kelvin, Lisa, Martin Nolf, Mehdi, Nadia, Nina, Nele, Nick, Oreste, Stijn, Wouter and all people who I have met in Belgium. You guys made my time in Belgium even better. I really hope to see you again. My Brazilian friends from Belgium, Ariane, Gabriela, Guido, Hianne, Hygor, Leandro, Pacheco, Pedro, Matheus, Radd, Samuel, Thales, Tiago and Tuane. Thanks for everything in Leuven, I could not have best flatmates. Thank Natália and Pascal for all support in Leuven. I wish you two all the best! Silvia, my favourite Italian in the world. You are my little sister and I really hope to see you and Riccardo again. K.U. Leuven for giving me a chance to study abroad and all technical support. My friends from Couchsurfing, thanks for being so welcoming and for sharing your time with me. I learnt a lot from you, specially my crazy friend Mati. One day you will win a Nobel Prize, but a really good one, Nobel Prize of Peace. Don’t forget me when it comes! Thank you guys and everyone else who made my academic life happier and enjoyable, I want to register here my deep gratitude. ABSTRACT The thesis was divided in four articles, in which three are related to in vitro propagation and how the microenvironmental conditions play on physiology and anatomy of B. zebrina. The last article is related to anatomical and physiological changes of B. zebrina under copper (Cu) excess stress. For all studies, B. zebrina plants were previously in vitro- established in MS medium. Plants were transferred to media at concentrations of 0%, 50%, 100%, 150% or 200% of the original salt concentration of MS medium. The media were prepared in two different consistencies, stationary liquid and 6 g L-1 agar. For in vitro rooting studies, the shoots grew in a medium supplemented with different sucrose concentrations. Soluble carbohydrates contents were assessed after the rooting. The in vitro multiplication of B. zebrina shoots is enhanced by using 200% of MS-salts concentration and liquid medium. The use of 15 g L-1 sucrose increased endogenous carbohydrate stocks and induced a good formation of the root systems on in vitro shoots. From these results, a second experiment was designed. B. zebrina side shoots were transferred to culture media containing 0.0, 15.0, 30.0, 45.0 or 60.0 g L-1 sucrose. Two different culture container sealing systems were tested: lids with a filter and a filter covered with PVC. At 45 days in vitro growth, B. zebrina plants were transplanted onto suitable soil mix and evaluated at 80 days growth in greenhouse. At 45 days in vitro and 80 days of acclimatization in the greenhouse, the biomass of plants was evaluated. Anatomical and physiological analysis were also performed on plants grown in vitro. Limited air exchange resulted in plantlets with anatomical and physiological disorders at the end of the in vitro period. The highest growth rate in the greenhouse was observed in plants previously propagated in unlimited gas exchange system and sugar-free medium. An environmental approach was proposed in the last study, in which copper was used. Anatomical and growth analysis were measured. Plants did not show any visible disturb, like necrosis on the leaves and all plants survived. Plants grown under 200 µM Cu showed anatomical changes that can help tolerating this metal, like high stomatal index and thicker cell wall in exodermis. Cu affected the leaf and root anatomy as well as on growth. B. zebrina tolerates high amounts of Cu. From the results it was possible to verify that microenvironmental conditions can change the growth, physiology and anatomy of B. zebrina during in vitro culture. In vitro technique showed a great potential on plant propagation of B. zebrina as well as it also presented an important tool for studies on plant physiology and anatomy. Keywords: Bromeliad. In vitro culture. Photoautotrophic growth. Plant anatomy. Plant physiology. RESUMO A tese foi dividida em quatro artigos, nos quais três estão relacionados a propagação in vitro e como as condições microambientas afetam a fisiologia e anatomia de B. zebrina. O último artigo está relacionado as modificações fisiologicas e anatômicas de B. zebrina sob estresse ao excesso de cobre (Cu). Para todos os estudos, plantas de B. zebrina foram previamente estabelecidas in vitro em meio MS. Plantas foram transferidas para meios nas concentrações de 0%, 50%, 100%, 150% e 200% da concentração original dos sais do meio MS. Os meios foram preparados em duas concentrações diferentes, líquido estacionário e com 6 g L-1 de ágar. Para os estudos de enraizamento in vitro, brotos foram cultivados em meios suplementado com diferentes concentrações de sacarose. Carboidratos solúveis foram analisados após o enraizamento. A multiplicação in vitro de B. zebrina é melhor com o uso de 200% dos sais MS e em meio líquido. O uso de 15 g L-1 de sacarose aumentou os estoques de carboidratos endógenos e induziram uma boa formação do sistema radicular dos brotos in vitro. A partir desses resultados, um segundo experimento foi delineado. Brotos laterais de B. zebrina foram transferidos para meios de cultura contendo 0,0; 15,0; 30,0; 45,0 ou 60,0 g L-1 de sacarose. Dois diferentes tipos de vedação dos frascos foram testados: tampas com um filtro e filtro coberto com plástico PVC. Aos 45 dias de cultivo in vitro, as plantas de B. zebrina foram transplantadas para uma mistura de solo e avaliadas aos 80 dias de cultivo em casa de vegetação. Aos 45 dias in vitro e 80 dias de aclimatização em casa de vegetação, a biomassa das plantas foi avaliada. Análises fisiológicas e anatômicas foram feitas nas plantas cultivadas in vitro.
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    Vol 40 Number 1 January/February/ March 2016 PUBLISHED BY: Editor - Derek Butcher. Assist Editor – Bev Masters Born 1977 and still offsetting!' COMMITTEE MEMBERS President: Adam Bodzioch 58 Cromer Parade Millswood 5034 Ph: 0447755022 Secretary: Bev Masters 6 Eric Street, Plympton 5038 Ph: 83514876 Vice president: Peter Hall Treasurer: Trudy Hollinshead Committee: Glenda Lee Penny Seekamp Julie Batty Dave Batty Sue Sckrabei Jeff Hollinshead Life members : Margaret Butcher, Derek Butcher, : Len Colgan, Adam Bodzioch Email address: Meetings Venue: Secretary – [email protected] Maltese Cultural Centre, Web site: http://www.bromeliad.org.au 6 Jeanes Street, Cultivar Register http://botu07.bio.uu.nl/bcg/bcr/index.php Beverley List for species names http://botu07.bio.uu.nl/bcg/taxonList.php http://botu07.bio.uu.nl/brom-l/ altern site http://imperialis.com.br/ Follow us on Face book Pots, Labels & Hangers - Small quantities available all meetings. Time: 2.00pm. For special orders/ larger quantities call Ron Masters on 83514876 Second Sunday of each month Exceptions –1st Sunday in March May, & August & 3rd Sunday September- no meeting in December or unless advised otherwise VISITORS & NEW MEMBERS WELCOME. Quesnelia ‘Tim Plowman’ MEETING & SALES 2016 DATES . 2/04/2016 & 3/04/2016 Sales , 10/4/2016 (Neutrog presentation) 1/5/2016 1st Sunday (Len’s Tillandsia presentation), 5/6/2016 1st Sunday (Workshop),10/07/2016 (Christmas in July & uncommon genera in SA), 7/08/2016 1st Sunday (winter brag) 18/09/2016 3rd Sunday (Mini’s) 9/10/2016 (Billbergia) 22/10/2016 & 23/10/2016 Sales 13/11/2016 130PM start, pup exchange, special afternoon tea – bring a plate of finger food to share, plant auction.
  • A Ética Ambiental No Código Florestal Brasileiro Sob a Ótica Dos Recursos Hídricos

    A Ética Ambiental No Código Florestal Brasileiro Sob a Ótica Dos Recursos Hídricos

    A ÉTICA AMBIENTAL NO CÓDIGO FLORESTAL BRASILEIRO SOB A ÓTICA DOS RECURSOS HÍDRICOS. A GESTÃO AMBIENTAL EM INSTITUIÇÕES DE ENSINO SUPERIOR BRASILEIRAS. A IMPLANTAÇÃO DE UM SISTEMA DE GESTÃO AMBIENTAL BASEADO NA NORMA ISO 14001/2004: UM ESTUDO DE CASO EM UMA FÁBRICA DE CONSERVAS. A PERCEPÇÃO DO RISCO AMBIENTAL, A ESCASSEZ DE ÁGUA E O CONFLITO DE INTERESSES NO VALE DO RIO DOS SINOS. ANÁLISE CITOTÓXICA DE EFLUENTE DOMÉSTICO PROVENIENTE DE DOIS SISTEMAS DE TRATAMENTO: LODO ATIVADO E FILTRO DE MACRÓFITAS EM FLUTUAÇÃO. ANÁLISE DA EVOLUÇÃO URBANA DE NOVO HAMBURGO COM ÊNFASE NAS ÁREAS DE RISCO. ANÁLISE DA INTERAÇÃO DOS SISTEMAS COMPLEXOS VERSUS O CONCEITO DA SUSTENTABILIDADE. ANÁLISE DOS INDICADORES DE SUSTENTABILIDADE CORPORATIVA. ANÁLISE PRELIMINAR DO DESENVOLVIMENTO ONTOGENÉTICO DE CYATHEA PHALERATA (CYATHEACEAE) SOB A INFLUÊNCIA DA LUZ. AS MUITAS ÁGUAS ENVOLVIDAS NUMA REFEIÇÃO. ASPECTOS E IMPACTOS AMBIENTAIS IDENTIFICADOS EM RESTAURANTES UNIVERSITÁRIOS DE UMA UNIVERSIDADE PÚBLICA FEDERAL. AVALIAÇÃO DA CAPACIDADE SELETIVA DA MEMBRANA DE NANOFILTRAÇÃO. AVALIAÇÃO DA QUALIDADE DA ÁGUA DE DOIS AFLUENTES DO RIO DOS SINOS, RS, ATRAVÉS DO USO DE BIOMARCADORES EM PEIXES. AVALIAÇÃO DAS MEMBRANAS ELETRODIÁLITICAS. AVALIAÇÃO INTEGRADA DA QUALIDADE AMBIENTAL EM UMA UNIDADE DE CONSERVAÇÃO NO MUNICÍPIO DE SÃO LEOPOLDO, RS, BRASIL CARACTERIZAÇÃO DE AEROSSÓIS EM ÁREAS URBANAS E SEMIURBANAS DA BACIA HIDROGRÁFICA DO RIO DOS SINOS. COMPORTAMENTO FENOLÓGICO E SAZONALIDADE DE GUAREA MACROPHYLLA VAHL EM FLORESTA ATLÂNTICA NO SUL DO BRASIL. CONSERVAÇÃO DE VRIESEA INCURVATA (BROMELIACEAE), ESPÉCIE ENDÊMICA DA FLORESTA ATLÂNTICA: AVALIAÇÃO DE NUTRIENTES PARA CULTURA IN VITRO. CURTUME: EFLUENTES, TOXICIDADE E NOVAS TECNOLOGIAS. DESENVOLVIMENTO DE MATERIAIS A PARTIR DA RECICLAGEM DE RESÍDUOS TÊXTEIS GERADOS PELA INDÚSTRIA CALÇADISTA.