Browsing by Author "Chahed, Abdessalem"
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- Determination of the subcellular localisation of GIP and NPP1 proteins in Phytophtora cinnamomi tagged with green fluorescent proteinPublication . Chahed, Abdessalem; Chatti, Noureddine; Costa, Rodrigo Arthur Fonseca; Sousa, Maria João; Choupina, AltinoP. cinnamomi is considered one of the world's worst invasive pathogens with a serious threat to a wide range of plant species throughout the world. These species cause several billions of dollars of damage on crop, ornamental, and native plants and so far no efficient solutions have been found to control the disease. A notable characteristic of the interaction between plants and their microbial pathogens is the secretion by both partners of proteins that are associated with attack, defense, and counterdefense thus it’s fundamental to improve our knowledge about these proteins with the intention of finding a solution to the disease affecting the Chestnut.
- Knockdown of Phytophthora cinnamomi gip gene by iRNAPublication . Chahed, Abdessalem; Costa, Rodrigo Arthur Fonseca; Sousa, Maria João; Choupina, AltinoInk Disease is considered one of the most important causes of the decline of chestnut orchards. The break in yield of Castanea sativa Mill is mainly caused by Phytophthora cinnamomi one of the most aggressive and widespread plant pathogen causing enormous economic losses and up to now no efficient treatments are available to fight these pathogens. Because of the importance of chestnut at economical and ecological levels especially in Portugal, it becomes essential to explore the molecular mechanisms that determine the interaction between Phytophthora species and host plants through the study of GIP protein produced by P.cinnamomi during the infection.
- Post-transcriptional gene silencing of glucanase inhibitor protein in Phytophthora cinnamomiPublication . Ferreira, Patrick; Chahed, Abdessalem; Estevinho, Leticia M.; Seixas, Natália; Costa, Rodrigo Arthur Fonseca; Choupina, AltinoInk disease is considered one of the most significant causes contributing to the decline of chestnut orchards. The reduced yield of Castanea sativa Mill can be attributed to two main species: Phytophthora cinnamomi and Phytophthora cambivora, with the first being the main pathogen responsible for ink disease in Portugal. P. cinnamomi is a highly aggressive and widely distributed plant pathogen, capable of infecting nearly 1000 host species. This oomycete causes substantial economic losses and is accountable for the decline of numerous plant species in Europe and worldwide. To date, no effective treatments are available to combat these pathogens. Given chestnut’s economic and ecological significance, particularly in Portugal, it is crucial to investigate the molecular mechanisms underlying the interaction between Phytophthora species and host plants. This can be achieved through the study of the glucanase inhibitor protein (GIP) produced by P. cinnamomi during infection. The technique of RNA interference (RNAi) was employed to suppress the GIP gene of P. cinnamomi. The resulting transformants, carrying the silenced gene, were used to infect C. sativa, allowing for the assessment of the effects of gene silencing on the plant’s phenotype. Additionally, bioinformatics tools predicted the secretion of GIP protein. The obtained results validate RNAi as a potential alternative tool for studying molecular factors and for controlling and managing P. cinnamomi.
- Silencing the gip gene of Phytophthora cinnamomi by iRNA and subcellular localization of GIP and NPP1 proteinsPublication . Chahed, Abdessalem; Chatti, Noureddine; Costa, Rodrigo Arthur Fonseca; Sousa, Maria João; Choupina, AltinoInk Disease is considered one of the most important causes of the decline of chestnut orchards. The break in yield of Castanea sativa Mill is mainly caused by Phytophthora cinnamomi one of the most aggressive and widespread plant pathogen causing enormous economic losses and up to now no efficient treatments are available to fight these pathogens. Because of the importance of chestnut at economical and ecological levels especially in Portugal, it becomes essential to explore the molecular mechanisms that determine the interaction between Phytophthora species and host plants through the study of proteins GIP and NPP1 produced by P.cinnamomi during the infection.
- Silencing the gip gene of Phytophthora cinnamomi by iRNA and subcellular localization of GIP and NPP1 proteinsPublication . Chahed, Abdessalem; Chatti, Noureddine; Costa, Rodrigo Arthur Fonseca; Sousa, Maria João; Choupina, AltinoInk Disease is considered one of the most important causes of the decline of chestnut orchards. The break-in yield of Castanea sativa Mill is mainly caused by Phytophthora cinnamomi one of the most aggressive and widespread plant pathogen causing enormous economic losses and up to now, no efficient treatments are available to fight these pathogens. Because of the importance of chestnut at economical and ecological levels especially in Portugal, it becomes essential to explore the molecular mechanisms that determine the interaction between Phytophthora species and host plants through the study of proteins GIP and NPP1 produced by P.cinnamomi during the infection.
- Silencing the gip gene of Phytophthora cinnamomi by iRNA and studying the subcellular localization of GIP and NPP1 proteinsPublication . Chahed, Abdessalem; Choupina, Altino; Sousa, Maria João; Chatti, NoureddineInk Disease is considered one of the most important causes of the decline of chestnut orchards. The break in yield of Castanea sativa Mill is caused by two species: Phytophthora cinnamomi and Phytophthora cambivora, being the first one the foremost pathogen of ink disease in Portugal. P. cinnamomi is one of the most aggressive and widespread plant pathogen with nearly 1,000 host species. This oomycete causes enormous economic losses and it is responsible for the decline of many plant species in Europe and worldwide. Up to now no efficient treatments are available to fight these pathogens. Because of the importance of chestnut at economical and ecological levels, especially in Portugal, it becomes essential to explore the molecular mechanisms that determine the interaction between Phytophthora species and host plants through the study of proteins GIP (glucanase inhibitor protein) and NPP1 (necrosis-inducing Phytophthora protein 1) produced by P. cinnamomi during the infection. The technique of RNA interference was used to knockdown the gip gene of P. cinnamomi. Transformants obtained with the silenced gene have been used to infect C. sativa, in order to determine the effect of gene silencing on the plant phenotype. To know more about the function of GIP and NPP1 involved in the mechanism of infection, the ORF’s of gip and npp1 genes have been cloned to the pTOR-eGFP vector for a future observation of P. cinnamomi transformants with fluorescent microscopy and determination of the subcellular localization. Moreover the prediction by bioinformatics tools indicates that both GIP and NPP1 proteins are secreted. The results allow to predict the secretory destination of both GIP and NPP1 proteins and confirm RNAi as a potential alternative biological tool in the control and management of P. cinnamomi. Keywords:
- Use of iRNA in the post-transcriptional gene silencing of necrosis-inducing Phytophthora protein 1(NPP1) in Phytophthora cinnamomiPublication . Pascoal‑Ferreira, Patrick; Chahed, Abdessalem; Costa, Rodrigo Arthur Fonseca; Branco, Iuliia; Choupina, AltinoPhytophthora cinnamomi is an Oomycetes associated with soil, this Oomycete is one of the most destructive species of Phytophthora, being responsible for the decline of more than 5000 ornamental, forest, or fruit plants. It can secrete a class of protein NPP1 (Phytophthora necrosis inducing protein 1), responsible for inducing necrosis in leaves and roots of plants, leading to their death. Objective This work will report the characterization of the Phytophthora cinnamomi NPP1 gene responsible for the infec tion of Castanea sativa roots and will characterize the mechanisms of interaction between Phytophthora cinnamomi and Castanea sativa, by gene silencing NPP1 from Phytophthora cinnamomi mediated by RNAi. Methods and results For silencing a part of the coding region of the NPP1 gene, was placed in the sense and antisense directions between an intron and ligated to the integrative vector pTH210. Cassette integration was confrmed by PCR and sequencing on the hygromycin-resistant Phytophthora cinnamomi transformants. Transformants obtained with the silenced gene was used to infect Castanea sativa. Conclusions Plants infected with these transformants showed a great reduction in disease symptoms, confrming iRNA as a potential alternative biological tool in the study of molecular factors, and in the control and management of Phytophthora cinnamomi.