A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to...A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) cryIAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptII) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.展开更多
Wheat （Triticum aestivum L.） is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution t...Wheat （Triticum aestivum L.） is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution to the increase in global food production over the past four decades （Dixon et al., 2009）. Wheat is produced on more than 18% of the arable land in the world, and is the most cultivated crop after maize and rice （FAOSTAT data, 2014）. Despite its global strategic significance, progress in genomic and genetic engineering research on wheat has lagged behind that on other major crops due to the difficulty of culturing tissues, and the complexity of its hexaploid genome. The first successful wheat trans- formation was achieved by particle bombardment （Vasil et al., 1992）. Since then additional transgenic wheat plants have been obtained by various transformation methods （Harwood, 2011）. Microprojectile bombardment is considered to be a promising method, since it is robust, versatile and relatively efficient in terms of gene delivery.展开更多
文摘A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) cryIAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptII) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.
基金funded by the Ministry of Agriculture of China(Nos.2014ZX0801003B and 2013ZX08002-004)
文摘Wheat （Triticum aestivum L.） is a major staple food crop worldwide. It is economically important because it can be grown in a wide range of climates and geographic regions, and it has made an enormous contribution to the increase in global food production over the past four decades （Dixon et al., 2009）. Wheat is produced on more than 18% of the arable land in the world, and is the most cultivated crop after maize and rice （FAOSTAT data, 2014）. Despite its global strategic significance, progress in genomic and genetic engineering research on wheat has lagged behind that on other major crops due to the difficulty of culturing tissues, and the complexity of its hexaploid genome. The first successful wheat trans- formation was achieved by particle bombardment （Vasil et al., 1992）. Since then additional transgenic wheat plants have been obtained by various transformation methods （Harwood, 2011）. Microprojectile bombardment is considered to be a promising method, since it is robust, versatile and relatively efficient in terms of gene delivery.
