Ice-cap. A high-throughput method for capturing plant tissue samples for genotype analysis
Ice-cap. A high-throughput method for capturing plant tissue samples for genotype analysis
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DOI：
10.1104/pp.104.040774
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发表时间：
2004-07-01
期刊：
影响因子：
7.4
通讯作者：
Krysan, P
中科院分区：
文献类型：
--
作者：
Krysan, P
高通量的基因筛查正迅速成为后基因组时代的标准研究工具。然而，目前存在的一个主要瓶颈限制了这种方法在植物科学中的应用。在目前的高通量管道中，限速步骤是必须手动采集活植物的组织样本，一次采集一个植物。在这篇文章中，我描述了一种从活的幼苗中采集组织样本的新方法，它消除了这一瓶颈。这种方法被命名为Ice-Cap，以反映冰被用来捕获组织样本的事实。种子的播种、幼苗的生长和组织的收获都是以96孔的形式进行的。我通过使用Ice-Cap收集的组织来对正在分离先前特征的突变的拟南芥幼苗群体进行基因分型，从而证明了该系统的实用性。因为组织的采集是以非破坏性的方式进行的，所以具有所需基因的植物可以被转移到土壤中并生长到成熟。我还证明了Ice-Cap可以用于分析水稻幼苗的基因组DNA。预计该方法将适用于多种不同植物的高通量筛选，成为进行分子标记辅助选择的有用技术。
High-throughput genotype screening is rapidly becoming a standard research tool in the post-genomic era. A major bottleneck currently exists, however, that limits the utility of this approach in the plant sciences. The rate-limiting step in current high-throughput pipelines is that tissue samples from living plants must be collected manually, one plant at a time. In this article I describe a novel method for harvesting tissue samples from living seedlings that eliminates this bottleneck. The method has been named Ice-Cap to reflect the fact that ice is used to capture the tissue samples. The planting of seeds, growth of seedlings, and harvesting of tissue are all performed in a 96-well format. I demonstrate the utility of this system by using tissue harvested by Ice-Cap to genotype a population of Arabidopsis seedlings that is segregating a previously characterized mutation. Because the harvesting of tissue is performed in a nondestructive manner, plants with the desired genotype can be transferred to soil and grown to maturity. I also show that Ice-Cap can be used to analyze genomic DNA from rice (Oryza saliva) seedlings. It is expected that this method will be applicable to high-throughput screening with many different plant species, making it a useful technology for performing marker assisted selection.