Concentración mínima inhibitoria de Kanamicina para callos de cuatro variedades colombianas de arroz

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Abstract

La Kanamicina (Km) es un antibiótico que se ha utilizado en la producción de plantas transgénicas de arroz como agen- te selectivo. En este trabajo se evaluó la sensibilidad al antibiótico para callos derivados de semillas sexuales en cuatro variedades colombianas de arroz (FEDEARROZ 2000, FEDEARROZ 50, FEDEARROZ 369 y CICA 8). Inicialmente se indujo la formación de callo en un medio específico a partir de semilla madura, obteniéndose porcentajes de callogénesis mayores al 90%. Posteriormente los callos son llevados a un medio de regeneración en el que se presentan variaciones importantes en el porcentaje de regeneración, entre 62% (F2000) y 11% (F369). Se ensayaron 7 concentraciones diferentes de Kanamicina, adicionadas al medio de regeneración: 0 mg/L, 100 mg/L, 200 mg/L, 300 mg/L, 400 mg/L, 500 mg/L y 600 mg/L. Se observó una respuesta diferencial al antibiótico. Para F2000 y F369 a partir de 500 mg/L no registraron regenerantes, mientras para F50 y CICA 8 fue desde 300 mg/L. En todos los casos se observaron regenerantes albinos.

Translated abstract

Kanamycin (Km) is an antibiotic that has been used in the production of transgenic rice plants as a selective agent. In this study, we evaluated the antibiotic sensitivity of seed derived callus in four varieties of colombian rice (FEDEARROZ 2000, FEDEARROZ 50, 369 and CICA FEDEARROZ 8). It was induced callus formation from mature sedd in a specific medium, it were resulting in percentages of callus formation greater than 90%. Later callus were subsequently taken to a regeneration medium, which are important variations in the rate of regeneration, between 62% (F2000) and 11% (F369). Seven different concentrations of kanamycin (0 mg/L, 100 mg/L, 200 mg/L, 300 mg/L, 400 mg/L, 500 mg/L and 600 mg/L) were tested added to the regeneration medium. It was observed differential response to the antibiotic. F369 to F2000 and from 500 mg/L showed no regenerating, while for F50 and CICA 8 was from 300 mg/L.

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Advances in selectable marker genes for plant transformation.

Natarajan Sakthivel, Isaac Sundar (2008)

Plant transformation systems for creating transgenics require separate process for introducing cloned DNA into living plant cells. Identification or selection of those cells that have integrated DNA into appropriate plant genome is a vital step to regenerate fully developed plants from the transformed cells. Selectable marker genes are pivotal for the development of plant transformation technologies because marker genes allow researchers to identify or isolate the cells that are expressing the cloned DNA, to monitor and select the transformed progeny. As only a very small portion of cells are transformed in most experiments, the chances of recovering transgenic lines without selection are usually low. Since the selectable marker gene is expected to function in a range of cell types it is usually constructed as a chimeric gene using regulatory sequences that ensure constitutive expression throughout the plant. Advent of recombinant DNA technology and progress in plant molecular biology had led to a desire to introduce several genes into single transgenic plant line, necessitating the development of various types of selectable markers. This review article describes the developments made in the recent past on plant transformation systems using different selection methods adding a note on their importance as marker genes in transgenic crop plants.

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Suitability of non-lethal marker and marker-free systems for development of transgenic crop plants: present status and future prospects.

G Ramkumar, P. Manimaran, S. Balachandran … (2011)

Genetically modified crops are one of the prudent options for enhancing the production and productivity of crop plants by safeguarding from the losses due to biotic and abiotic stresses. Agrobacterium-mediated and biolistic transformation methods are used to develop transgenic crop plants in which selectable marker genes (SMG) are generally deployed to identify 'true' transformants. The commonly used SMG obtained from prokaryotic sources when employed in transgenic plants pose risks due to their lethal nature during selection process. In the recent past, some non-lethal SMGs have been identified and used for selection of transformants with increased precision and high selection efficiency. Considering the concerns related to bio-safety of the environment, it is desirable to remove the SMG in order to maximize the commercial success through wide adoption and public acceptance of genetically modified (GM) food crops. In this review, we examine the availability, and the suitability of wide range of non-lethal selection markers and elimination of SMG methods to develop marker-free transgenics for achieving global food security. As the strategies for marker-free plants are still in proof-of-concept stage, adaptation of new genomics tools for identification of novel non-lethal marker systems and its application for developing marker-free transgenics would further strengthen the crop improvement program.