Efficient screening of transgenic plant lines for ecological research

Plants stably transformed to manipulate the expression of genes mediating ecological performance have profoundly altered research in plant ecology. Agrobacterium-mediated transformation remains the most effective method of creating plants harboring a limited number of transgene integrations of low complexity. For ecological/physiological research, the following requirements must be met: 1) the regenerated plants should have the same ploidy level as the corresponding wild type plant and 2) contain a single transgene copy in a homozygous state; 3) the T-DNA must be completely inserted without vector backbone sequence and all its elements functional; and 4) the integration should not change the phenotype of the plant by interrupting chromosomal genes or by mutations occurring during the regeneration procedure. The screening process to obtain transformed plants that meet the above criteria is costly and time consuming and an optimized screening procedure is presented. We developed a flow ch...

通过稳定转化以调控介导生态功能的基因表达的植物，已深刻改变了植物生态学研究。农杆菌介导的转化（Agrobacterium-mediated transformation）仍是构建具有数量有限、复杂度低的转基因整合植株的最有效方法。对于生态/生理学研究，需满足以下要求：1）再生植株应与对应的野生型植株具有相同的倍性水平；2）含有纯合状态的单拷贝转基因；3）T-DNA需完整插入且无载体骨架序列，其所有元件均功能正常；4）整合过程不应通过中断染色体基因或再生过程中发生的突变改变植株表型。获得满足上述标准的转化植株的筛选过程既昂贵又耗时，因此本文提出了一种优化的筛选流程。我们开发了一种流式...