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尽管应用基因转化进行果树品种改良已日益引起重视,但是在受体的应用和转化方法上还存在着诸多困难。一方面,大多数果树尚不能从细胞或原生质体再生成完整植株,即使少数已可以再生的果树树种,也并非众多品种都能再生成功,而是存在着明显的基因型差异性。同时,还有再生植株童期过长的问题。另一方面,目前在植物基因转化中常用的两种方法即DNA直接吸入法和农杆菌介导的载体法,若以细胞或原生质体为受体,不仅存在再生困难的问题,而且再生过程费时长;若以叶盘、愈伤组织或珠心组织等为受体,既需要在转化后除去农杆菌,又需要排除转化与非转化组织的嵌合性。这些因素都大大地限制了基因转化在果树中的应用。因此,根据多年生果树的生长特点,建立一种适用的基因转化技术已成当务之急。本文采用农杆菌介导的附体腋芽转化-离体扩繁鉴定的方法,成功地将GUS基因转入沙田柚。结果证明这是一种简单、快速、高效的基因转化方法。
Although the application of genetic transformation to improve the fruit tree species has drawn increasing attention, there are still many difficulties in the application and transformation methods of the receptor. On the one hand, most fruit trees are still unable to regenerate intact plants from cells or protoplasts. Even for a few fruit tree species that can be regenerated, not many species can be successfully regenerated, but there are obvious genotypic differences. In the meantime, there is the problem of prolonged childhood in regenerated plants. On the other hand, two methods commonly used in plant gene transformation, that is, direct DNA inhalation and Agrobacterium-mediated vector methods, not only have the problem of regeneration difficulty, but also take the expense of the regeneration process if cells or protoplasts are used as receptors Time; if the leaf disc, calli or callus tissue as the recipient, both need to remove agrobacteria after transformation, but also need to rule out the conversion and non-transformed tissue chimerism. These factors have greatly limited the application of gene transformation in fruit trees. Therefore, based on the perennial growth characteristics of fruit trees, establishing a suitable gene transformation technology has become a top priority. In this paper, Agrobacterium-mediated transformation of axillary bud annexing - in vitro propagation identification method, the successful transfer of GUS gene into Shatian pomelo. The results prove that this is a simple, rapid and efficient gene transformation method.