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首页> 外文期刊>Journal of Biotechnology >Gene stacking in 1-year-cycling APETALA1 citrus plants for a rapid evaluation of transgenic traits in reproductive tissues
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Gene stacking in 1-year-cycling APETALA1 citrus plants for a rapid evaluation of transgenic traits in reproductive tissues

机译：基因堆叠在1年周期的APETALA1柑橘植物中，用于快速评估生殖组织中的转基因性状

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Rapid flowering is crucial to perform functional genomic studies to investigate reproductive biology characteristics and fruit quality-related traits in fruit trees. However, long generation cycles of woody plants considerably delay this evaluation. Through genetic transformation, juvenile periods can be significantly shortened by overexpression of flower meristem-identity genes. Transgenic APETALA1 (AP1) citrus plants behave as rapid-cycling trees, since 1-year-old seedlings promptly show precocious flowering and fruiting. By transgene stacking into these short-generation AP1 and nptII/GUS-positive plants, expression of novel transgenes could theoretically be examined as quickly as 1 year after retransformation. Establishment of the selection and regeneration conditions for the production of retransformed individuals with marker genes is detailed in this communication. Hpt and bar genes were used as the second selectable marker genes. PCR and Southern blot analyses confirmed the recovery of retransformed shoots. AP1 transcript accumulation and GUS and GFP expression were assessed in leaves, and flowers and fruit organs of rapid-cycling retransformed lines, respectively, as early as 1 year after plant generation and during three consecutive years, demonstrating that the principle of stable transgene stacking on early-fruiting transgenic trees is feasible.
机译：快速开花对于进行功能基因组研究以调查果树的生殖生物学特性和与果实品质相关的性状至关重要。但是，木本植物的长生成周期大大延迟了该评估。通过遗传转化，花分生组织同一性基因的过表达可以大大缩短少年时期。转基因的APETALA1（AP1）柑橘类植物表现为快速循环的树，因为1岁的幼苗迅速显示出早熟的开花结果。通过将转基因堆叠到这些短代AP1和nptII / GUS阳性植物中，理论上可以在重新转化后的1年内尽快检查新型转基因的表达。本交流详细介绍了建立具有标记基因的转化个体的选择和再生条件。 Hpt和bar基因被用作第二选择标记基因。 PCR和Southern印迹分析证实了重新转化的芽的恢复。早在植物产生后的第一年和连续三年中，分别评估了快速循环转化株系的叶片，花和果实器官中的AP1转录物积累和GUS和GFP表达，证明了稳定转基因的原理早结果转基因树是可行的。

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