Agrobacterium -mediated transformation of Brachypodium distachyon inbred line Bd21 with two binary vectors containing hy
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J. Crop Sci. Biotech. 2011 (December) 14 (4) : 233 ~ 238 DOI No. 10.1007/s12892-011-0080-9 RESEARCH ARTICLE
Agrobacterium-Mediated Transformation of Brachypodium distachyon Inbred Line Bd21 with Two Binary Vectors Containing Hygromycin Resistance and GUS Reporter Genes Man Bo Lee, Woong Bae Jeon, Dae Yeon Kim, Odgerel Bold, Min Jeong Hong, Yong Jin Lee, Jae Han Park, Yong Weon Seo* College of Life Sciences and Biotechnology, Korea University, Seoul, 136-713, Korea Received: August 9, 2011 / Revised: September 9, 2011 / Accepted: September 16, 2011 Ⓒ Korean Society of Crop Science and Springer 2011
Abstract Brachypodium distachyon (Brachypodium) is a novel model plant for structural and functional genomic studies of temperate grasses. Brachypodium as a model plant has many favorable features, such as small size, small genome, short life cycle, self-fertility, and simple growth requirements. The genome sequence of the standard line Bd21 has been released and genomic resources have been developed.It is imperative to develop a method for efficient Agrobacterium-mediated Brachypodium transformation. Yellowish and compact embryogenic calli derived from immature embryos of the Bd21 were transformed with the Agrobacterium strain AGL1. Seven- and nine-week-old calli were used for transformation with Agrobacterium carrying either pCAMBIA 1301 and pCAUGH. Transformation efficiencies were assessed through histochemical GUS assay. The efficiency of transformation with pCAMBIA 1301 (based on the number of callus lines producing GUS-detected plantlets and the number of calli used for transformation) reached 20.1% (7-week-old calli) and 1.7% (9-week-old calli), and with pCAUGH (based on the number of GUS-detected plantlets and the number of regenerants) 90 and 87% for 7- and 9-week-old calli, respectively. High selection pressure was obtained by using pCAUGH, which is preferred for saving labor and time consumption during the callus selection. Key words: Agrobacterium, Brachypodium distachyon, immature embryo, pCAMBIA 1301, pCAUGH
Introduction Brachypodium distachyon has been highly spotlighted as a novel model system for the study of the Triticeae in the past decade. Arabidopsis thaliana as a generalized model plant has contributed to the advancement of modern plant biology (Vogel and Bragg 2008). Although Arabidopsis is still powerful system, Arabidopsis is not an ideal model for the grasses (Keller and Feuillet 2000). There are biological differences such as cell wall composition between dicots and monocots because they diverged nearly 150 million years ago (Vogel and Bragg 2008). There were efforts to develop rice as a model plant more suitable for Poaceae (Goff 1999). Rice is a semi-aquatic tropical grass that diverged from forage grasses and temperate grains nearly 50 million years ago (Gaut 2002). Since riceis a semiYong Weon Seo ( ) E-mail: [email protected] Tel: +82-2-3290-3005 / Fax: +82-2-3290-3501 The Korean Society of Crop Science
aquatic tropical grass, it may not suitable for other cereals such as wheat, barley,
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