Improved Genetic Transformation of Sugarcane (Saccharum spp.) Embryogenic Callus Mediated by Agrobacterium tumefaciens.

Autor: Basso MF; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., da Cunha BADB; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Ribeiro AP; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Martins PK; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., de Souza WR; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., de Oliveira NG; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Nakayama TJ; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Augusto das Chagas Noqueli Casari R; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Santiago TR; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Vinecky F; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Cançado LJ; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., de Sousa CAF; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., de Oliveira PA; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., de Souza SACD; Agronomic Institute of Campinas (IAC), Ribeirão Preto, São Paulo, Brazil., Cançado GMA; The Joint Research Unit for Genomics Applied to Climate Change (UMIP GenClima), National Center for Agricultural Informatics (CNPTIA), Brazilian Agricultural Research Corporation (EMBRAPA), Campinas, São Paulo, Brazil., Kobayashi AK; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil., Molinari HBC; Genetics and Biotechnology Laboratory, National Center for Agroenergy Research (CNPAE), Brazilian Agricultural Research Corporation (EMBRAPA), Brasília, Distrito Federal, Brazil.
Jazyk: angličtina
Zdroj: Current protocols in plant biology [Curr Protoc Plant Biol] 2017 Sep; Vol. 2 (3), pp. 221-239.
DOI: 10.1002/cppb.20055
Abstrakt: Sugarcane (Saccharum spp.) is a monocotyledonous semi-perennial C4 grass of the Poaceae family. Its capacity to accumulate high content of sucrose and biomass makes it one of the most important crops for sugar and biofuel production. Conventional methods of sugarcane breeding have shown several limitations due to its complex polyploid and aneuploid genome. However, improvement by biotechnological engineering is currently the most promising alternative to introduce economically important traits. In this work, we present an improved protocol for Agrobacterium tumefaciens-mediated transformation of commercial sugarcane hybrids using immature top stalk-derived embryogenic callus cultures. The callus cultures are transformed with preconditioned A. tumefaciens carrying a binary vector that encodes expression cassettes for a gene of interest and the bialaphos resistance gene (bar confers resistance to glufosinate-ammonium herbicide). This protocol has been used to successfully transform a commercial sugarcane cultivar, SP80-3280, highlighting: (i) reduced recalcitrance and oxidation; (ii) high yield of embryogenic callus; (iii) improved selection; and (iv) shoot regeneration and rooting of the transformed plants. Altogether, these improvements generated a transformation efficiency of 2.2%. This protocol provides a reliable tool for a routine procedure for sugarcane improvement by genetic engineering. © 2017 by John Wiley & Sons, Inc.
(Copyright © 2017 John Wiley & Sons, Inc.)
Databáze: MEDLINE
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