High transformation efficiency in plum (Prunus domestica L.):: a new tool for functional genomics studies in Prunus spp.
High transformation efficiency in plum (Prunus domestica L.):: a new tool for functional genomics studies in Prunus spp.
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DOI：
10.1007/s11032-008-9200-8
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发表时间：
2008-11-01
影响因子：
3.1
通讯作者：
Scorza, Ralph
中科院分区：
文献类型：
--
作者：
Petri, Cesar;Webb, Kevin;Scorza, Ralph
采用改良的农杆菌介导法对李（Prunus arctica L.）在共培养培养基中添加2，4-D，培养下胚轴切片。该方法将转化效率提高至先前报道的10倍（42%），下胚轴切片产生转基因植物的平均效率为25%。优化方案中每个步骤的时间，在温室中在约6个月内产生自根转基因植物。为了测试其在功能基因组研究中的实用性的系统，我们开发了两个发夹结构使用桃（P. persica）八氢番茄红素脱氢酶（PDS）基因的片段。当A.将具有这些构建体的根癌农杆菌用于靶向转录后基因沉默（PTGS），约50%的转化的李子是敲除PDS基因的植物。我们在这里报告的简单和有效的李转化协议可以很容易地用于功能基因组学研究李属，蔷薇科和其他木本植物一般。
An improved Agrobacterium-mediated transformation protocol for plum (Prunus domestica L.) hypocotyl slices was developed based on the addition of 2,4-D to the co-cultivation medium. This method increased transformation efficiency up to 10 x (42%) over previous reports with an average efficiency of 25% of hypocotyl slices producing transgenic plants. Timing of each step in the protocol was optimized producing self-rooted transgenic plants in the greenhouse in approximately 6 months. In order to test the system for its utility in functional genomic studies, we developed two hairpin constructs using a fragment of the peach (P. persica) Phytoene desaturase (PDS) gene. When A. tumefaciens with these constructs was used for targeted post-transcriptional gene silencing (PTGS), approximately 50% of the transformed plums were knockout PDS gene plants. The easy and efficient plum transformation protocol that we report here can be readily used for functional genomics studies in Prunus specifically, and Rosaceae and other woody species in general.