Highly efficient transformation of zonal (Pelargonium x hortorum) and scented (P. capitatum) geraniums via Agrobacterium tumefaciens using leaf discs
Highly efficient transformation of zonal (Pelargonium x hortorum) and scented (P. capitatum) geraniums via Agrobacterium tumefaciens using leaf discs
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DOI：
10.1016/j.plantsci.2005.04.014
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发表时间：
2005-09
期刊：
影响因子：
5.2
通讯作者：
A. Hassanein;E. Chevreau;N. Dorion
中科院分区：
文献类型：
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作者：
A. Hassanein;E. Chevreau;N. Dorion
首次建立了天竺葵和黄花天竺葵的高效快速转化体系。使用两种携带含有npt II和uidA基因的pFAJ 3000载体的根癌农杆菌菌株。3天的共培养期提高了转化效率。接种前的预培养步骤对间接再生的大麦有正效应，对直接再生的鸭茅有负效应。EHA 101的最佳接种时间为20 min，而LBA 4404的最佳接种时间为10 min。通常，发现LBA 4404比EHA 101更有效地转化天竺葵属。以产生转基因植株的外植体数为基础，大麦的转化效率达到73%，以从独立事件中分离的转基因植株数为基础，大麦的转化效率达到146%。对柱头草的转化效率分别达到100%和346%。鸭茅和鸭茅的再生过程分别持续了13周和17周。PCR分析证实转化，组织化学分析检测GUS表达。流式细胞仪分析显示，只有P. x hortorum的转基因克隆中有一些四倍化。该过程现在可以有效地用于改善天竺葵和/或研究基因功能。
Highly efficient and rapid transformation system was established, for the first time, using leaf discs of Pelargonium x hortorum and P. capitatum. Two Agrobacterium tumefaciens strains harboring pFAJ3000 vector containing nptII and uidA genes were used. A 3 days of co-cultivation period enhanced transformation efficiency. Pre-culture step before inoculation had a positive effect on indirectly regenerated P. x hortorum and a negative effect on directly regenerated P. capitatum. A 20min inoculation duration was optimum with EHA101 whereas 10min were sufficient for LBA4404. Generally, LBA4404 was found more efficient than EHA101 to transform both Pelargonium. The transformation efficiency for P. x hortorum reached 73% based on the number of explants producing transgenic plants and 146% based on the number of transgenic plants isolated from independent events. For P. capitatum, transformation efficiency was up to 100 and 346%, respectively. The regeneration process lasted 13 and 17 weeks, respectively, for P. capitatum and P. x hortorum. The transformation was confirmed by PCR analysis and GUS expression was tested by histochemical assay. Flow cytometry analysis revealed some tetraploidization among transgenic clones of P. x hortorum only. The process can be efficiently used now to improve geraniums and/or study gene function.