Genetic transformation of 'Valencia' sweet orange with the cecropin MB39 gene

O objetivo deste trabalho foi obter plantas transgênicas de laranja 'Valência' com o gene cecropin MB39 controlado pelo promotor do gene da fenilalanina-amônia-liase de citros, visando a expressão gênica específica nos vasos do xilema. A transformação genética foi realizada via Agrobacterium tumefaciens por meio do co-cultivo de segmentos de epicótilo. Onze plantas transgênicas foram identificadas por PCR, pela amplificação do fragmento esperado de 189 pb, as quais foram aclimatizadas em casa de vegetação. A integração do transgene foi confirmada em três plantas pela análise de transferência de Southern.The objective of this work was to produce 'Valencia' sweet orange transgenic plants with the cecropin MB39 gene controlled by a phenylalanine ammonia-lyase gene promoter from citrus in order to direct gene expression in xylem vessels. The genetic transformation was mediated by Agrobacterium tumefaciens with the co-culture of epicotyl segments. Eleven transgenic plants were selected by PCR with the amplification of a 189 bp fragment, which were acclimatized to greenhouse. The integration of the transgene was confirmed in three plants by Southern blot analysis

Towards the identification of flower-specific genes in Citrus spp

Citrus sinensis is a perennial woody species, for which genetic approaches to the study of reproductive development are not readily amenable. Here, the usefulness of the CitEST Expressed Sequence Tag (EST) database is demonstrated as a reliable new resource for identifying novel genes exclusively related to Citrus reproductive biology. We performed the analysis of an EST dataset of the CitEST Project containing 4,330 flower-derived cDNA sequences. Relying on bioinformatics tools, sequences exclusively present in this flower-derived sequence collection were selected and used for the identification of Citrus putative flower-specific genes. Our analysis revealed several Citrus sequences showing significant similarity to conserved genes known to have flower-specific expression and possessing functions related to flower metabolism and/or reproductive development in diverse plant species. Comparison of the Citrus flower-specific sequences with all available plant peptide sequences unraveled 247 unique transcripts not identified elsewhere within the plant kingdom. Additionally, 49 transcripts, for which no biological function could be attributed by means of sequence comparisons, were found to be conserved among plant species. These results allow further gene expression analysis and possibly novel approaches to the understanding of reproductive development in Citrus

Towards the identification of flower-specific genes in Citrus spp

Citrus sinensis is a perennial woody species, for which genetic approaches to the study of reproductive development are not readily amenable. Here, the usefulness of the CitEST Expressed Sequence Tag (EST) database is demonstrated as a reliable new resource for identifying novel genes exclusively related to Citrus reproductive biology. We performed the analysis of an EST dataset of the CitEST Project containing 4,330 flower-derived cDNA sequences. Relying on bioinformatics tools, sequences exclusively present in this flower-derived sequence collection were selected and used for the identification of Citrus putative flower-specific genes. Our analysis revealed several Citrus sequences showing significant similarity to conserved genes known to have flower-specific expression and possessing functions related to flower metabolism and/or reproductive development in diverse plant species. Comparison of the Citrus flower-specific sequences with all available plant peptide sequences unraveled 247 unique transcripts not identified elsewhere within the plant kingdom. Additionally, 49 transcripts, for which no biological function could be attributed by means of sequence comparisons, were found to be conserved among plant species. These results allow further gene expression analysis and possibly novel approaches to the understanding of reproductive development in Citrus.761768Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Transformação genética de laranja 'Valência' com o gene cecropin MB39 Genetic transformation of 'Valencia' sweet orange with the cecropin MB39 gene

O objetivo deste trabalho foi obter plantas transgênicas de laranja 'Valência' com o gene cecropin MB39 controlado pelo promotor do gene da fenilalanina-amônia-liase de citros, visando a expressão gênica específica nos vasos do xilema. A transformação genética foi realizada via Agrobacterium tumefaciens por meio do co-cultivo de segmentos de epicótilo. Onze plantas transgênicas foram identificadas por PCR, pela amplificação do fragmento esperado de 189 pb, as quais foram aclimatizadas em casa de vegetação. A integração do transgene foi confirmada em três plantas pela análise de transferência de Southern.The objective of this work was to produce 'Valencia' sweet orange transgenic plants with the cecropin MB39 gene controlled by a phenylalanine ammonia-lyase gene promoter from citrus in order to direct gene expression in xylem vessels. The genetic transformation was mediated by Agrobacterium tumefaciens with the co-culture of epicotyl segments. Eleven transgenic plants were selected by PCR with the amplification of a 189 bp fragment, which were acclimatized to greenhouse. The integration of the transgene was confirmed in three plants by Southern blot analysis

A genetic framework for flowering-time pathways in Citrus spp.

Floral transition is one the most drastic changes occurring during the life cycle of a plant. The shoot apical meristem switches from the production of leaves with associated secondary shoot meristems to the production of flower meristems. This transition is abrupt and generally irreversible, suggesting it is regulated by a robust gene regulatory network capable of driving sharp transitions. The moment at which this transition occurs is precisely determined by environmental and endogenous signals. A large number of genes acting within these pathways have been cloned in model herbaceous plants such as Arabidopsis thaliana. In this paper, we report the results of our search in the Citrus expressed sequence tag (CitEST) database for expressed sequence tags (ESTs) showing sequence homology with known elements of flowering-time pathways. We have searched all sequence clusters in the CitEST database and identified more than one hundred Citrus spp sequences that codify putative conserved elements of the autonomous, vernalization, photoperiod response and gibberelic acid-controlled flowering-time pathways. Additionally, we have characterized in silico putative members of the Citrus spp homologs to the Arabidopsis CONSTANS family of transcription factors