Plant Tissue Culture and Genetic Transformation

Somatic Embryogenesis in Recalcitrant Plants

by Antonio Andrade-Torres

Laura Yesenia Solís-Ramos, Antonio Andrade-Torres, Luis Alfonso Sáenz Carbonell, Carlos M. Oropeza Salín and Enrique Castaño de la Serna. 2012. 25. Somatic Embryogenesis in Recalcitrant Plants. In: Ken-ichi Sato (Ed). Embryogenesis. InTech Open. Pp. 597-618. ISBN 978-953-51-0466-7.
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Somatic embryogenesis is the process by which somatic cells, under induction conditions, generate embryogenic cells,... more Somatic embryogenesis is the process by which somatic cells, under induction conditions, generate embryogenic cells, which go through a series of morphological and biochemical changes that result in the production of bipolar structure without vascular connection with the original tissue. Somatic embryogenesis with a low frequency of chimeras, a high number of regenerates and a limited level of somaclonal variation is more attractive than organogenesis as a plant regeneration system (e.g., in genetic transformation, in vitro mutagenesis and selection). However there are several factors that influence the initiation of somatic embryogenesis in plants. Most of the important crops and grasses are recalcitrant for in vitro culturing, which hampers the development of reliable regeneration techniques. This document is focused in the somatic embryogenesis of recalcitrant plants, showing the particular cases of two plant species: habanero chili (Capsicum chinense Jacq.) and coconut palm (Cocos nucifera L.).

Transient genetic transformation of embryogenic callus of Cocos nucifera

by Antonio Andrade-Torres

Andrade-Torres, A.*, C. Oropeza, L. Sáenz, T. González-Estrada, J. E. Ramírez-Benítez, K. Becerril, J. L. Chan, and L. C. Rodríguez-Zapata. 2011. Transient genetic transformation of embryogenic callus of Cocos nucifera L. Biologia. 66 (5): 790-800. ISSN: 0006-3088.

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* pregunta por este artículo a: myxo23_@hotmail.com o lcrz@cicy.mx

PGFPGUS Plus, a New Binary Vector for Gene Expression Studies and Optimising Transformation Systems In Plants

by Claudia Vickers

A binary vector containing two reporter gene cassettes has been developed. This vector is ideal for optimising new... more A binary vector containing two reporter gene cassettes has been developed. This vector is ideal for optimising new plant transformation systems. Following optimisation, one of the reporter genes can be replaced with a gene of interest; the second can be used as a marker to confirm transgenic lines, and to estimate locus number and determine zygosity. This allows simple, efficient and economical screening for homozygous single-insert lines and azygous controls.

A synthetic xylanase as a novel reporter in plants

by Claudia Vickers

Transient gene expression assays are often used to screen promoters before stable transformation. Current transient... more Transient gene expression assays are often used to screen promoters before stable transformation. Current transient quantification methods have several problems, including a lack of reporter gene stability and expense. Here we report a synthetic, codon-optimised xylanase gene (sXynA) as a reporter gene for quantitative transient analyses in plants. Azurine-crosslinked xylan (AZCL-xylan) was used as a substrate for assaying xylanase activity. The enzymatic nature of the protein allows for sensitive assays at the low levels of transgene protein found in transiently transformed tissue extracts. The xylanase (XYN) protein is stable, activity slopes are linear over long time periods and assays are cost-effective. Coupled with the GUSPlus reporter gene, the XYN reporter allows sensitive and accurate quantification of gene control sequences in transient expression systems

Indirect somatic embryogenesis and morphohistological analysis in Capsicum chinense

by Antonio Andrade-Torres

Solís-Ramos, L.Y.*, S. Nahuat-Dzib, A. Andrade-Torres, F. Barredo-Pool, T. González-Estrada and E. Castaño. 2010. Indirect somatic embryogenesis and morphohistological analysis in Capsicum chinense Jacq. Biologia. 65 (3): 504-511. ISSN: 0006-3088.
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* Pregunta por este artículo al email: catleyasp@hotmail.com

Endogenous GUS-like activity in Capsicum chinense Jacq

by Antonio Andrade-Torres

Solís-Ramos, L.Y., T. González-Estrada, A. Andrade-Torres, G. Godoy-Hernández y E. Castaño. 2010. Endogenous Gus-like activity in Capsimun chinense Jacq. Electronic Journal of Biotechnology. 13 (4). ISSN: 0717-3458