Efficiency of Agrobacterium rhizogenes-mediated root transformation of Parasponia and Trema is temperature dependent
Efficiency of Agrobacterium rhizogenes-mediated root transformation of Parasponia and Trema is temperature dependent
复制标题
DOI：
10.1007/s10725-012-9734-y
复制
发表时间：
2012-12-01
影响因子：
4.2
通讯作者：
Geurts, Rene
中科院分区：
文献类型：
--
作者：
Cao, Qingqin;Op den Camp, Rik;Geurts, Rene
拟松属树木是唯一能与根瘤菌形成固氮根瘤的非豆科植物。基于其在豆科植物（豆科）中的分类地位，这两个谱系很可能独立地获得了这种共生能力。因此，Parasponia形成了一个桥接物种，以了解这种共生关系背后的进化限制。然而，缺乏关键的技术来转基因Parasponia严重阻碍了对这些物种的研究。我们采用发根农杆菌创造复合Parasponia andersonii植物海港转基因根。在这里，我们提供了一个优化的协议，感染P. andersonii以及它的非共生姐妹种Trema tomentosa与A。发根我们表明，转换效率是温度依赖性。尽管这两种物种的最佳生长温度是28摄氏度，但当与A.发根发生在21摄氏度。使用这种优化的方案可以获得高达80%的转化效率。这些强大的转化平台将提供一个强大的工具来解开副棘孢-根瘤菌共生。
Parasponia trees are the only non-legume species that form nitrogen-fixing root nodules with rhizobium. Based on its taxonomic position in relation to legumes (Fabaceae), it is most likely that both lineages have gained this symbiotic capacity independently. Therefore, Parasponia forms a bridging species to understand the evolutionary constraints underlying this symbiosis. However, absence of key technologies to genetically modify Parasponia seriously impeded studies on these species. We employed Agrobacterium rhizogenes to create composite Parasponia andersonii plants that harbour transgenic roots. Here, we provide an optimized protocol to infect P. andersonii as well as its non-symbiotic sister species Trema tomentosa with A. rhizogenes. We show that the transformation efficiency is temperature dependent. Whereas the optimal growth temperature for both these species is 28 degrees C, the transformation is most efficient when co-cultivation with A. rhizogenes occurs at 21 degrees C. Using this optimized protocol up to 80 % transformation efficiency can be obtained. These robust transformation platforms will provide a strong tool to unravel the Parasponia-rhizobium symbiosis.