Sujatha Mulpuri, Visarada K B R S
Directorate of Oilseeds Research, Rajendranagar, Hyderabad, India.
Methods Mol Biol. 2013;940:27-44. doi: 10.1007/978-1-62703-110-3_3.
Particle bombardment/biolistic delivery is a very popular method of genetic transformation of diverse targets including cells and intact tissues. Delivery of DNA through particle bombardment is genotype and species independent, nevertheless, an efficient protocol for large-scale generation of transgenic plants through embryogenic tissues with a high (≥80%) shoot regeneration efficiency is a prerequisite. Young embryogenic tissues or multiple shoot buds in early stages of induction are the most suited target tissues for recovery of transgenic plants. We describe the protocol for delivery of foreign genes using particle delivery system (Biorad gene gun, PDS-1000/He) in to the meristematic tissues of embryonic axes derived from mature seeds of castor. With the optimized physical and biological parameters, putative transformants were obtained at a frequency of 1.4% through particle gun bombardment of castor embryo axes. Also, transformation of embryogenic calli of sorghum using particle inflow gun (PIG) is described.
粒子轰击/生物弹道传递是一种非常流行的遗传转化方法，可用于多种靶标，包括细胞和完整组织。通过粒子轰击传递DNA不受基因型和物种限制，然而，通过具有高（≥80%）芽再生效率的胚性组织大规模生成转基因植物的有效方案是一个先决条件。幼胚性组织或诱导早期的多个芽是恢复转基因植物最适合的靶组织。我们描述了使用粒子传递系统（伯乐基因枪，PDS-1000/He）将外源基因传递到蓖麻成熟种子衍生的胚轴分生组织中的方案。通过优化物理和生物学参数，通过粒子枪轰击蓖麻胚轴以1.4%的频率获得了推定转化体。此外，还描述了使用粒子流入枪（PIG）对高粱胚性愈伤组织进行转化的过程。
