Systematic approaches to using the FOX hunting system to identify useful rice genes
Systematic approaches to using the FOX hunting system to identify useful rice genes
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DOI：
10.1111/j.1365-313x.2008.03733.x
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发表时间：
2009-03-01
期刊：
影响因子：
7.2
通讯作者：
Matsui, Minami
中科院分区：
文献类型：
--
作者：
Kondou, Youichi;Higuchi, Mieko;Matsui, Minami
异位基因表达或功能获得法的优点是，一旦知道了基因的功能，就可以通过转化将基因转移到许多不同的植物上。我们以前报道了一种名为Fox Hunting的方法，该方法涉及在拟南芥中异位表达全长cDNA，以系统地产生功能获得突变体。这项技术最有利于为分析有用的植物基因功能而产生异源基因资源。作为初始模型，我们产生了23000多个表达水稻fl-cDNAs的独立拟南芥转基因株系(水稻福克斯拟南芥系)。拟南芥代代时间短，转化频率快，可以快速分析水稻基因的功能。我们对福克斯拟南芥的形态、光合作用、元素积累、色素积累、激素谱、次生代谢产物、抗病、耐盐性、紫外线信号、高光耐性和耐热胁迫等方面的变化进行了筛选。福克斯拟南芥品系所显示的一些突变表型是由于在拟南芥中没有同源基因的水稻基因的表达所致。这一结果表明水稻fl-cDNA可以用于在拟南芥中引入新的基因功能。此外，这些发现表明，可以利用拟南芥作为异源寄主来分析水稻的基因功能。该技术为分析植物在异源寄主中的基因功能和利用异源基因资源进行植物改良提供了框架。
Ectopic gene expression, or the gain-of-function approach, has the advantage that once the function of a gene is known the gene can be transferred to many different plants by transformation. We previously reported a method, called FOX hunting, that involves ectopic expression of Arabidopsis full-length cDNAs in Arabidopsis to systematically generate gain-of-function mutants. This technology is most beneficial for generating a heterologous gene resource for analysis of useful plant gene functions. As an initial model we generated more than 23 000 independent Arabidopsis transgenic lines that expressed rice fl-cDNAs (Rice FOX Arabidopsis lines). The short generation time and rapid and efficient transformation frequency of Arabidopsis enabled the functions of the rice genes to be analyzed rapidly. We screened rice FOX Arabidopsis lines for alterations in morphology, photosynthesis, element accumulation, pigment accumulation, hormone profiles, secondary metabolites, pathogen resistance, salt tolerance, UV signaling, high light tolerance, and heat stress tolerance. Some of the mutant phenotypes displayed by rice FOX Arabidopsis lines resulted from the expression of rice genes that had no homologs in Arabidopsis. This result demonstrated that rice fl-cDNAs could be used to introduce new gene functions in Arabidopsis. Furthermore, these findings showed that rice gene function could be analyzed by employing Arabidopsis as a heterologous host. This technology provides a framework for the analysis of plant gene function in a heterologous host and of plant improvement by using heterologous gene resources.