A rapid and efficient transformation protocol for the grass Brachypodium distachyon
A rapid and efficient transformation protocol for the grass Brachypodium distachyon
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DOI：
10.1007/s00299-004-0889-5
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发表时间：
2005-03-01
影响因子：
6.2
通讯作者：
Nielsen, KK
中科院分区：
文献类型：
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作者：
Christiansen, P;Andersen, CH;Nielsen, KK
报道了一种快速有效的微弹轰击介导的禾本科植物二穗短柄草（Brachypodium distachyon）的转化方案，该短柄草是禾本科植物功能基因组学中水稻的替代模式植物。通过含有uidA（编码β-葡萄糖醛酸酶）和bar（编码草丁膦乙酰转移酶）基因的构建体转化来自未成熟胚的胚性愈伤组织，并在组织培养的所有阶段使用双丙氨膦（bialaphos）（一种非选择性除草剂）作为选择剂。平均转化效率为5.3%，并观察到高达14%的单轰击转化效率。从轰击胚性愈伤组织到收获转基因T-1种子的时间框架对于所使用的二倍体和两个四倍体加入物分别为29周和25周。由于二倍体的种子到种子的生命周期为19周，四倍体的为15周，因此我们的B。二穗草转化系统允许在1年内测试T-0和T-1代以及生产T-2种子。
A fast and efficient microprojectile bombardment-mediated transformation protocol is reported for the grass species Brachypodium distachyon, a proposed alternative model plant to Oryza sativa for functional genomics in grasses. Embryogenic calli derived from immature embryos were transformed by a construct containing the uidA ( coding for beta-glucuronidase) and bar ( coding for phosphinothricin acetyl transferase) genes, and bialaphos, a non-selective herbicide, was used as the selection agent throughout all phases of the tissue culture. Average transformation efficiencies of 5.3% were achieved, and for single bombardments transformation efficiencies of up to 14% were observed. The time frame from the bombardment of embryogenic callus to the harvesting of transgenic T-1 seeds was 29 weeks and 25 weeks for the diploid and two tetraploid accessions used, respectively. Since the seed-to-seed life cycle is 19 weeks for the diploid and 15 weeks for the tetraploid accessions, our B. distachyon transformation system allows testing of both the T-0 and the T-1 generation as well as production of T-2 seeds within 1 year.