Doutorado em Genética e Melhoramento
URI Permanente para esta coleção
Nível: Doutorado
Ano de início: 2013
Conceito atual na CAPES: 5
Ato normativo: Homologado pelo CNE/CES Parecer nº 250/2014, portaria nº 187 de 06/03/2015, publicado no DOU de 09/03/2015 seção 1, página 11
Periodicidade de seleção: Semestral
Área(s) de concentração: Genética e Melhoramento
Url do curso: https://geneticaemelhoramento.ufes.br/pt-br/pos-graduacao/PPGGM/detalhes-do-curso?id=1416
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Navegando Doutorado em Genética e Melhoramento por Autor "Clarindo, Wellington Ronildo"
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- ItemEmbriogênese somática indireta e poliploidia no gênero Coffea : base e aplicação(Universidade Federal do Espírito Santo, 2017-07-14) Sanglard, Natália Arruda; Ferreira, Márcia Flores da Silva; Clarindo, Wellington Ronildo; Mendonça, Maria Andréia Côrrea; Werner, Elias Terra; Soares, Fernanda Aparecida Ferrari; Fontes, Milene Miranda PraçaIndirect somatic embryogenesis (ISE) is one of the tissue culture applications that allow the maintenance, propagation and generation of new germplasms. Thus, from the establishment of ISE in four Coffea with different ploidy levels (diploids Coffea canephora Pierre ex Froehner and Coffea eugenioides Moore; allotriploid "Híbrido de Timor" – HT, and allotetraploid Coffea arabica L.), the aims of the present study were: (a) to verify the relation between karyotype features (chromosome number, ploidy level and nuclear DNA content) and ISE; and (b) doubling the chromosome number of the allotriploid HT 'CIFC 4106'. Under the same in vitro conditions, the four Coffea varieties differed from each other during all steps of the ISE. The allopolyploids provided the highest mean number of friable embryogenic calli (FEC), in relatively short time, and visually exhibiting more pronounced cell proliferation. FEC of C. arabica resulted in the highest mean number of mature cotyledonary somatic embryos, followed by HT and C. eugenioides with the same mean value, and C. canephora with the lowest amount. Regarding the ISE data, chromosome number, ploidy level and nuclear 2C value, in vitro responses in Coffea were related to ploidy level. From established ISE, chromosome doubling of the allotriploid HT 'CIFC 4106', an anorthoploid with 2n = 3x = 33 chromosomes, was conducted by associating this in vitro system with colchicine treatment. A total of 65 plantlets were regenerated from the FEC treated with colchicine (0.5, 1.5 or 2.5 mM) for 96 h. Independently of the applied xv colchicine concentration, a rate of 49.3% of hexaploids (2n = 6x = 66 chromosomes, 2C = 4.20 pg) was obtained. In addition, the established strategy (ISE/colchicine) solved the main bottlenecks of in vitro chromosome doubling: low rate of polyploids, high number of mixoploids and high mortality. This study provided new data on the basics of ISE, understanding and contributing with a new strategy of chromosome doubling. In addition, a new Coffea germplasm, the HT 'CIFC 4106' hexaploids, was generated.
- ItemMUDANÇAS E DIFERENÇAS EPIGENÉTICAS NA ORGANOGÊNESE E EMBRIOGÊNSE SOMÁTICA EM PASSIFLORA: INFLUÊNCIA DO EXPLANTE E DAS CONDIÇÕES IN VITRO(Universidade Federal do Espírito Santo, 2021-08-26) Leite, Cristiana Torres; Fontes, Milene Miranda Praca; https://orcid.org/0000000177389518; http://lattes.cnpq.br/1372644511398549; https://orcid.org/0000-0002-7036-4200; http://lattes.cnpq.br/2070518226531886; Clarindo, Wellington Ronildo; https://orcid.org/0000000288268620; http://lattes.cnpq.br/0339970047422716; Oliveira, Stéfanie Cristina de; https://orcid.org/0000-0001-8350-2601; http://lattes.cnpq.br/6404885772178652; Werner, Elias Terra; https://orcid.org/0000-0001-7781-4342; http://lattes.cnpq.br/3268068963656927; Oliveira, Joao Paulo de Morais; https://orcid.org/0000-0001-9780-9807; http://lattes.cnpq.br/8146032394984908The development of in vitro plants can be influenced by several factors, among them the explant source, the cultivation conditions and the global methylation of the genomic DNA. It is known that in species of Passiflora L. genus, different morphogenic responses are observed under the same conditions in vitro. Therefore, other factors may influence the response, such as the genetic, physiological and epigenetic aspects of the explant. We aim here to establish the in vitro morphogenesis process for three species of the genus Passiflora in order to verify and compare the influence of explant types, cultivation conditions and the global level of 5-methylcytosine. Our results showed that in Passiflora miniata the regeneration was through indirect somatic embryogenesis pathway, with the highest number of somatic embryos formed from immature zygotic embryos, in the absence of growth regulators and activated charcoal. Furthermore, the overall level of 5-methylcytosine decreased in the regeneration stage. In Passiflora cristalina and Passiflora foetida, the morphogenic pathway was indirect organogenesis, with the highest bud formation triggered by the use of mature zygotic embryos, the presence of growth regulators and activated charcoal, as well as the increase in the global level of 5-methylcytosine in the regeneration stage. From these different responses, we evidenced the influence of the developmental stage of the zygotic embryo, the use of growth regulators and activated charcoal and, for the first time, the global level of 5-methylcytosine during the in vitro development of Passiflora species.
- ItemOrigin of the allotriploid "híbrido de timor" through a karyotype comparison with its coffea ancestors(Universidade Federal do Espírito Santo, 2017-12-21) Oliveira, Stéfanie Cristina de; Fontes, Milene Miranda Praça; https://orcid.org/0000-0001-7738-9518; http://lattes.cnpq.br/1372644511398549; Soares, Fernanda Aparecida Ferrari; https://orcid.org/0000-0002-7198-1577; http://lattes.cnpq.br/5855273025512026; Clarindo, Wellington Ronildo; https://orcid.org/0000-0002-8826-8620; http://lattes.cnpq.br/0339970047422716; https://orcid.org/0000-0001-8350-2601; http://lattes.cnpq.br/6404885772178652; Karsburg, Isane Vera; https://orcid.org/0000-0002-9637-0449; http://lattes.cnpq.br/4736928411585458; Carvalho, Guilherme Mendes de Almeida; https://orcid.org/0009-0003-2724-1955; http://lattes.cnpq.br/8143011123794868Among the Coffea, there is a natural hybrid denominated “Híbrido de Timor” (HT), found on Timor Island in 1927. HT 'CIFC 4106', which represents the first HT plant, possesses 2n = 3x = 33 chromosomes and 1C DNA value equal 1C = 2.10 pg. Chromosomal number, nuclear 1C DNA content and geographical evidences support a possible allotriploid origin from fusion of a reduced reproductive cell of Coffea arabica (2n = 4x = 44) and Coffea canephora (2n = 2x = 22). C. arabica, another allopolyploid, accumulates studies that seek to unravel its progenitors. Molecular and karyotype data suggests that this true allotetraploid was formed from a reduced reproductive cell of C. canephora (CC) and another C. eugenioides (EE) followed by a polyploidization event. In this way, C. arabica genome is represented as C aC a E a E a . Based on this evidences, we formulate the following hypothesis: HT ‘CIFC 4106’ genome is CCa E a ? The present study aimed to cytogenetically characterize C. eugenioides, C. canephora, C. arabica and HT 'CIFC 4106'. The combination of morphometric data, flow and image cytometries and 5S rDNA fluorescente in situ hybridization (FISH) expanded knowledge about the evolutive origin and genome structure of HT ‘CIFC 4106’. HT 'CIFC 4106' karyogram evidenced chromosomes group delimited according to the class and DNA value similar. Based on these karyotype features was possible show the presence of two identical genomes in HT 'CIFC 4106', possibly of C. canephora (CC) and a distinct genome (C. eugenioides, E). HT 'CIFC 4106' chromosomes presented class, DNA content identical to the chromosomes of C. eugenioides, C. canephora and C. arabica. 5S rDNA sites were detected in two HT ‘CIFC 4106’ chromosomes. Distribution patterns of 5S markings found in C. eugenioides and C. canephora revealed in this study corroborate with HT ‘CIFC 4106’ CCa E a genome hypothesis. The 15 distribution of the 5S rDNA sites found in three Coffea allied to the chromosomal DNA content and class shows chromosomes similar to C. eugenioides and C. canephora in HT ‘CIFC 4106’. In addition, the data reveal possible structural chromosomal rearrangements along the evolution of HT ‘CIFC 4106’
- ItemThe epigenetic and physiologic changes as a trigger for indirect somatic embryogenesis in Coffea canephora(Universidade Federal do Espírito Santo, 2019-07-15) Silva, Paulo Marcos Amaral; Clarindo, Wellington Ronildo; https://orcid.org/0000000288268620; http://lattes.cnpq.br/0339970047422716; https://orcid.org/; http://lattes.cnpq.br/2853274010679824; Fontes, Milene Miranda Praca; https://orcid.org/0000000177389518; http://lattes.cnpq.br/1372644511398549; Mendonca, Maria Andreia Correa; https://orcid.org/0000-0002-3798-3309; http://lattes.cnpq.br/3412339192891083; Sanglard, Natalia Arruda; https://orcid.org/0000-0003-3219-8616; http://lattes.cnpq.br/7809043431671687; Carrijo, Tatiana Tavares; https://orcid.org/0000000283660738; http://lattes.cnpq.br/0479854777557016Indirect somatic embryogenesis (ISE) establishment for Coffea species started in the 1970s. Since then, intraspecific variations in the morphogenic pathway have been reported, despite the meticulous standardization of the environmental conditions in vitro. Several authors have suggested that these variations are the result of genetic, epigenetic and/or physiological events, highlighting the need for investigations to know the respective causes. Along these lines, this study aimed to investigate and describe, for the first time, the changes in and influence of epigenetic and physiological aspects during cell proliferation and somatic embryo regeneration from cell aggregate suspensions of Coffea canephora. The cell proliferation step was characterized by increase in cell mass in all subcultures; relatively low mean values of global 5-methylcytosine (5-mC%), abscisic acid (ABA) and indole-3-acetic acid (IAA); high mean value of 1-aminocyclopropane 1-carboxylic acid (ACC, an ethylene precursor); and increases followed by decreases in spermidine (Spd, a polyamine) levels. With regard to the in vitro conditions, these aspects promoted the cell proliferation, showing that the epigenome and metabolome are fundamental for the first morphogenic moment of ISE. In turn, the second ISE moment is correlated with epigenetic and physiological changes. The competence acquisition, determination and cell differentiation steps were marked by increases in mean values of 5-mC%, IAA and ABA and decreases in ACC and Spd, evincing that these epigenetic and physiological changes are the triggers for regeneration and maturation of somatic embryos. Therefore, dynamic and coordinated epigenetic and physiological changes directly affect the two morphogenic moments of C. canephora ISE.