Casas A M, Kononowicz A K, Zehr U B, Tomes D T, Axtell J D, Butler L G, Bressan R A, Hasegawa P M
Center for Plant Environmental Stress Physiology, Purdue University, West Lafayette, IN 47907.
Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11212-6. doi: 10.1073/pnas.90.23.11212.
Transgenic sorghum plants have been obtained after microprojectile bombardment of immature zygotic embryos of a drought-resistant sorghum cultivar, P898012. DNA delivery parameters were optimized based on transient expression of R and C1 maize anthocyanin regulatory elements in scutellar cells. The protocol for obtaining transgenic plants consists of the delivery of the bar gene to immature zygotic embryos and the imposition of bialaphos selection pressure at various stages during culture, from induction of somatic embryogenesis to rooting of regenerated plantlets. One in about every 350 embryos produced embryogenic tissues that survived bialaphos treatment; six transformed callus lines were obtained from three of the eight sorghum cultivars used in this research. Transgenic (T0) plants were obtained from cultivar P898012 (two independent transformation events). The presence of the bar and uidA genes in the T0 plants was confirmed by Southern blot analysis of genomic DNA. Phosphinothricin acetyltransferase activity was detected in extracts of the T0 plants. These plants were resistant to local application of the herbicide Ignite/Basta, and the resistance was inherited in T1 plants as a single dominant locus.
通过对耐旱高粱品种P898012的未成熟合子胚进行微粒轰击，已获得转基因高粱植株。基于R和C1玉米花青素调控元件在盾片细胞中的瞬时表达，对DNA导入参数进行了优化。获得转基因植株的方案包括将bar基因导入未成熟合子胚，并在从体细胞胚发生诱导到再生小植株生根的培养过程中的各个阶段施加双丙氨膦选择压力。大约每350个胚中有一个产生了能在双丙氨膦处理下存活的胚性组织；从本研究中使用的八个高粱品种中的三个获得了六个转化愈伤组织系。从品种P898012获得了转基因（T0）植株（两个独立的转化事件）。通过对基因组DNA的Southern印迹分析证实了T0植株中bar和uidA基因的存在。在T0植株的提取物中检测到了草丁膦乙酰转移酶活性。这些植株对除草剂Ignite/Basta的局部施用具有抗性，并且该抗性在T1植株中作为单个显性位点遗传。
