Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis.

Guignardia citricarpa is the causal agent of Citrus Black Spot (CBS), an important disease in Citriculture. Due to the expressive value of this activity worldwide, especially in Brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control CBS. This work describes for the first time an efficient and successful methodology for genetic transformation of G. citricarpa mycelia, which generated transformants expressing the gene encoding for the gfp (green fluorescent protein) and also their interaction with citrus plant. Mycelia of G. citricarpa were transformed via Agrobacterium tumefaciens, which carried the plasmid pFAT-gfp, contains the genes for hygromycin resistance (hph) as well as gfp. The optimization of the agrotransformation protocol was performed testing different conditions (type of membrane; inductor agent concentration [acetosyringonee - AS] and cocultivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200µM of AS and 96 h as cocultivation time. High mitotic stability (82 %) was displayed by transformants using Polymerase Chain Reaction (PCR) technique to confirm the hph gene insertion. In addition, the presence of gfp was observed inside mycelia by epifluorescence optical microscopy. This technique easy visualization of the behaviour of the pathogen interacting with the plant for the first time, allowing future studies on the pathogenesis of this fungus. The establishment of a transformation method for G. citricarpa opens a range of possibilities and facilitates the study of insertional mutagenesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plant-pathogen interaction

Tensor product representation of Kothe-Bochner spaces and their dual spaces

We provide a tensor product representation of Kothe-Bochner function spaces of vector valued integrable functions. As an application, we show that the dual space of a Kothe-Bochner function space can be understood as a space of operators satisfying a certain extension property. We apply our results in order to give an alternate representation of the dual of the Bochner spaces of p-integrable functions and to analyze some properties of the natural norms that are defined on the associated tensor products.First and third authors are supported by grant MTM201453009-P of the Ministerio de Economia y Competitividad (Spain). Second and fourth authors are supported by grant MTM2012-36740-C02-02 of the Ministerio de Economia y Competitividad (Spain).Calabuig, JM.; Jiménez Fernández, E.; Juan Blanco, MA.; Sánchez Pérez, EA. (2016). Tensor product representation of Kothe-Bochner spaces and their dual spaces. Positivity. 20(1):155-169. https://doi.org/10.1007/s11117-015-0347-3S155169201Bochner, S.: Integration von Funktionen, deren Werte die Elemente eines Vectorraumes sind. Fundamenta Mathematicae 20, 262–276 (1933)Calabuig, J.M., Delgado, O., Juan, M.A., Sánchez, E.A.: Pérez, On the Banach lattice structure of $$L^1_w$$ L w 1 of a vector measure on a $$\delta$$ δ -ring. Collect. Math. 65, 6567–85 (2014)Calabuig, J.M., Delgado, O., Sánchez Pérez, E.A.: Factorizing operators on Banach function spaces through spaces of multiplication operators. J. Math. Anal. Appl. 364(1), 88–103 (2010)Calabuig, J.M., Gregori, P., Sánchez, E.A.: Pérez, Radon-Nikodým derivatives for vector measures belonging to Köthe function spaces. J. Math. Anal. Appl. 348, 469–479 (2008)Cerdà, J., Hudzik, H., Mastyło, M.: Geometric properties of Köthe-Bochner spaces. Math. Proc. Cambridge Philos. Soc. 120(3), 521–533 (1996)Chakraborty, N.D., Basu, S.: Spaces of p-tensor integrable functions and related Banach space properties. Real Anal. Exchange 34, 87–104 (2008)Chakraborty, N.D., Basu, S.: Integration of vector-valued functions with respect to vector measures defined on $$\delta$$ δ -rings. Ill. J. Math. 55(2), 495–508 (2011)Defant, A., Floret, K.: Tensor norms and operator ideals. North-Holland, Amsterdam (1993)Delgado, O., Juan, M.A.: Representation of Banach lattices as $$L^{1}_{w}$$ L w 1 spaces of a vector measure defined on a $$\delta -$$ δ - ring. Bull. Belgian Math. Soc. 19, 239–256 (2012)Diestel, J., Uhl, J.J.: Vector measures. Am. Math. Soc, Providence (1977)Dobrakov, I.: On integration in Banach spaces, VII. Czechoslovak Math. J. 38, 434–449 (1988)García-Raffi, L.M., Jefferies, B.: An application of bilinear integration to quantum scattering. J. Math. Anal. Appl. 415, 394–421 (2014)Gregori Huerta, P.: Espacios de medidas vectoriales. Thesis, Universidad de Valencia, ISBN:8437060591 (2005)Jefferies, B., Okada, S.: Bilinear integration in tensor products. Rocky Mt. J. Math. 28, 517–545 (1998)Lewis, D.R.: On integrability and summability in vector spaces. Ill. J. Math. 16, 294–307 (1972)Lin, P.-K.: Köthe-Bochner function spaces. Birkhauser, Boston (2004)Lindenstrauss, J., Tzafriri, L.: Classical Banach spaces II. Springer, Berlin (1979)Okada, S., Ricker, W.J., Sánchez Pérez, E.A.: Optimal Domains and integral extensions of operators acting in function spaces. Operator Theory Advances and Applications, vol. 180. Birkhäuser, Basel (2008)Pallu de La Barriére, R.: Integration of vector functions with respect to vector measures. Studia Univ. Babes-Bolyai Math. 43, 55–93 (1998)Rodríguez, J.: On integration of vector functions with respect to vector measures. Czechoslovak Math. J. 56, 805–825 (2006

Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis

Guignardia citricarpa is the causal agent of Citrus Black Spot (CBS), an important disease in Citriculture. Due to the expressive value of this activity worldwide, especially in Brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control CBS. This work describes for the first time an efficient and successful methodology for genetic transformation of G. citricarpa mycelia, which generated transformants expressing the gene encoding for the gfp (green fluorescent protein) and also their interaction with citrus plant. Mycelia of G. citricarpa were transformed via Agrobacterium tumefaciens, which carried the plasmid pFAT-gfp, contains the genes for hygromycin resistance (hph) as well as gfp. The optimization of the agrotransformation protocol was performed testing different conditions (type of membrane; inductor agent concentration [acetosyringonee - AS] and cocultivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200µM of AS and 96 h as cocultivation time. High mitotic stability (82 %) was displayed by transformants using Polymerase Chain Reaction (PCR) technique to confirm the hph gene insertion. In addition, the presence of gfp was observed inside mycelia by epifluorescence optical microscopy. This technique easy visualization of the behaviour of the pathogen interacting with the plant for the first time, allowing future studies on the pathogenesis of this fungus. The establishment of a transformation method for G. citricarpa opens a range of possibilities and facilitates the study of insertional mutagenesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plant-pathogen interaction.Made available in DSpace on 2017-08-10T23:53:57Z (GMT). No. of bitstreams: 1 cpamtFerreira187861462013.pdf: 2546936 bytes, checksum: 2e6c44d693e8209961e5bcc1042c2b08 (MD5) Previous issue date: 2014-02-0