Agrobacterium-mediated transformation of Brachypodium distachyon through embryogenic calli derived from immature embryos

Acta Prataculturae Sinica ›› 2010, Vol. 19 ›› Issue (5): 9-16.

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Agrobacterium-mediated transformation of Brachypodium distachyon through embryogenic calli derived from immature embryos

WU Xue-li¹, LIU Jin-xing², Klaus K Nielsen³,YANG Zhi-min¹,GAO Cai-xia^2,3

1.College of Horticulture, Nanjing Agricultural University, Nanjing 210095,China;
2.Institute of
Genetics and Developmental Biology Chinese Academy of Sciences, State Key Laboratory of
Plant Cell and Chromosome Engineering, Beijing 100101,China;
3.Research Division,
DLF-TRIFOLIU M A/S 466060, Denmark

Received:2009-10-10 Online:2010-05-25 Published:2010-10-20

Abstract

Abstract: A very efficient transformation system, using Agrobacterium-mediated transformation, has been developed for the production of transgenic plants of Brachypodium distachyon. Embryogenic calli initiated from immature embryos were transformed with Agrobacterium tumefaciens strain AGL1 carrying the binary vector pDM805, coding for the phosphinothricin acetyltransferase (bar) and β-glucuronidase (uidA) genes. Bialaphos was used as the selective agent throughout all phases of tissue culture. Transformation efficiency was strongly influenced by genotype, size of immature embryos and age of embryogenic callus. An average transformation efficiency of 38.5% was obtained using 5 to 8 week-old embryogenic callus as transformation targets and the highest transformation efficiency achieved was 43.1%. The existence and integration of the bar gene in transgenic plants was confirmed by PCR (polymerase chain reaction).

CLC Number:

WU Xue-li, LIU Jin-xing, Klaus K Nielsen,YANG Zhi-min,GAO Cai-xia. Agrobacterium-mediated transformation of Brachypodium distachyon through embryogenic calli derived from immature embryos[J]. Acta Prataculturae Sinica, 2010, 19(5): 9-16.

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Agrobacterium-mediated transformation of Brachypodium distachyon through embryogenic calli derived from immature embryos

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