ABSTRACT

Mango (Mangifera indica L.) is one of the most popular tropical fruits in the world owing to its rich taste, flavor, color, production volume and diverse end usage. Conventional mango breeding practices are unable to withstand the demand for improved varieties as it is time consuming and requires heavy investment. However, problems associated with traditional plant breeding can be curtailed through genetic transformation. Nevertheless, major limitation of transgenic development has been its recalcitrant nature toward tissue culture practices involving latent microbial infection, phenol exudation, etc. This opens wide scope for tissue culture-independent in planta transformation approaches These strategies have proved to be easy to execute and cost effective in producing large number of transformants. One such apical meristem targeted in planta approach was successfully exploited to demonstrate its utility in transforming a tree species. Mango variety Amrapali was transformed with two visual marker gene vectors GFP::hptII in pCAMBIA1302 and GUS::nptII in pCAMBIA2301 individually, to demonstrate its amenability. Preliminary confirmations identified 65.0% of GFP and 57.14% of GUS plants to be transformed. Further, molecular characterization of these primary transformants demonstrated transgene integration at genomic and transcript level in some of the plants. This established protocol holds good for functional gene validation and knock in/out studies and aid in mango improvement programs.

摘要

芒果（Mangifera indica L.）以其丰富的口感、风味、色泽、产量和用途广泛而成为世界上最受欢迎的热带水果之一。传统的芒果育种方法无法满足对改良品种的需求，因为它既费时又需要大量投资。然而，与传统植物育种相关的问题可以通过遗传转化来减少。然而，转基因发展的主要限制是其对涉及潜伏微生物感染、苯酚渗出等组织培养实践的顽固性质。这为植物中的组织培养独立开辟了广阔的范围转化方法 这些策略已被证明易于执行，并且在产生大量转化体方面具有成本效益。一个这样的以植物方法为目标的顶端分生组织被成功地利用来证明它在改造树种中的效用。芒果品种 Amrapali 在 pCAMBIA1302 中分别用两个视觉标记基因载体GFP::hptII和pCAMBIA2301 中的GUS::nptII进行转化，以证明其适应性。初步确认确定了 65.0% 的GFP和 57.14% 的GUS要转化的植物。此外，这些初级转化体的分子特征表明转基因在一些植物的基因组和转录物水平上进行了整合。该既定协议适用于功能基因验证和敲入/敲出研究，并有助于芒果改良计划。