D'Halluin K, Bonne E, Bossut M, De Beuckeleer M, Leemans J
Plant Genetic Systems N.V., Gent, Belgium.
Plant Cell. 1992 Dec;4(12):1495-505. doi: 10.1105/tpc.4.12.1495.
In this paper, we describe the transformation of regenerable maize tissues by electroporation. In many maize lines, immature zygotic embryos can give rise to embryogenic callus cultures from which plants can be regenerated. Immature zygotic embryos or embryogenic type I calli were wounded either enzymatically or mechanically and subsequently electroporated with a chimeric gene encoding neomycin phosphotransferase (neo). Transformed embryogenic calli were selected from electroporated tissues on kanamycin-containing media and fertile transgenic maize plants were regenerated. The neo gene was transmitted to the progeny of kanamycin-resistant transformants in a Mendelian fashion. This showed that all transformants were nonchimeric, suggesting that transformation and regeneration are a single-cell event. The maize transformation procedure presented here does not require the establishment of genotype-dependent embryogenic type II callus or cell suspension cultures and facilitates the engineering of new traits into agronomically relevant maize inbred lines.
在本文中,我们描述了通过电穿孔法对可再生玉米组织进行转化的过程。在许多玉米品系中,未成熟的合子胚能够产生胚性愈伤组织培养物,并从中再生出植株。未成熟的合子胚或I型胚性愈伤组织通过酶解或机械损伤处理,随后用编码新霉素磷酸转移酶(neo)的嵌合基因进行电穿孔。从在含卡那霉素的培养基上的电穿孔组织中筛选出转化的胚性愈伤组织,并再生出可育的转基因玉米植株。neo基因以孟德尔方式传递给抗卡那霉素转化体的后代。这表明所有转化体均为非嵌合体,这意味着转化和再生是单细胞事件。本文介绍的玉米转化方法不需要建立依赖基因型的II型胚性愈伤组织或细胞悬浮培养物,并且便于将新性状导入具有农艺相关性的玉米自交系中。