期刊
PLANT GROWTH REGULATION
卷 68, 期 3, 页码 459-465出版社
SPRINGER
DOI: 10.1007/s10725-012-9734-y
关键词
Parasponia; Trema; Agrobacterium transformation; Symbiosis; Transgenic root; Composite plant
资金
- Dutch Science Foundation (NWO) [VIDI 864.06.007]
- Dutch Graduate School of Experimental Plant Sciences
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.
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