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首页> 外文期刊>Planta: An International Journal of Plant Biology >Agrobacterium tumefaciens and Agrobacterium rhizogenes transformed roots of the parasitic plant Triphysaria versicolor retain parasitic competence
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Agrobacterium tumefaciens and Agrobacterium rhizogenes transformed roots of the parasitic plant Triphysaria versicolor retain parasitic competence

机译：根癌农杆菌和发根农杆菌转化的寄生植物杂色Triphysaria versicolor的根保留了寄生能力

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Parasitic plants in the Orobanchaceae invade roots of neighboring plants to rob them of water and nutrients. Triphysaria is facultative parasite that parasitizes a broad range of plant species including maize and Arabidopsis. In this paper we describe transient and stable transformation systems for Triphysaria versicolor Fischer and C. Meyer. Agrobacterium tumefaciens and Agrobacterium rhizogenes were both able to transiently express a GUS reporter in Triphysaria seedlings following vacuum infiltration. There was a correlation between the length of time seedlings were conditioned in the dark prior to infiltration and the tissue type transformed. In optimized experiments, nearly all of the vacuum infiltrated seedlings transiently expressed GUS activity in some tissue. Calluses that developed from transformed tissues were selected using non-destructive GUS staining and after several rounds of in vivo GUS selection, we recovered uniformly staining GUS calluses from which roots were subsequently induced. The presence and expression of the transgene in Triphysaria was verified using genomic PCR, RT PCR and Southern hybridizations. Transgenic roots were also obtained by inoculating A. rhizogenes into wounded Triphysaria seedlings. Stable transformed roots were identified using GUS staining or fluorescent microscopy following transformation with vectors containing GFP, dsRED or EYFP. Transgenic roots derived from both A. tumefaciens and A. rhizogenes transformations were morphologically normal and developed haustoria that attached to and invaded lettuce roots. Transgenic roots also remained competent to form haustoria in response to purified inducing factors. These transformation systems will allow an in planta assessment of genes predicted to function in plant parasitism.
机译：Orobanchaceae中的寄生植物入侵附近植物的根，抢劫它们的水分和养分。 Triphysaria是兼性寄生虫，可寄生包括玉米和拟南芥在内的多种植物。在本文中，我们描述了Triphysaria versicolor Fischer和C. Meyer的瞬时和稳定转化系统。真空渗入后，根癌土壤杆菌和发根土壤杆菌都能够在Tri虫幼苗中瞬时表达GUS报告基因。渗透前在黑暗中调理幼苗的时间长度与转化的组织类型之间存在相关性。在优化的实验中，几乎所有的真空渗透幼苗在某些组织中都瞬时表达了GUS活性。使用非破坏性GUS染色选择从转化组织发育的愈伤组织，经过几轮体内GUS选择后，我们回收了均匀染色的GUS愈伤组织，随后从中诱导了根。使用基因组PCR，RT PCR和Southern杂交验证了Triphysaria中转基因的存在和表达。还可以通过将发根农杆菌接种到受伤的紫phy幼苗中来获得转基因根。用含有GFP，dsRED或EYFP的载体转化后，使用GUS染色或荧光显微镜鉴定稳定的转化根。从根癌农杆菌和发根农杆菌转化获得的转基因根在形态上是正常的，并且发育了附着于并侵入莴苣根的酒糟。转基因的根也仍然能够响应纯化的诱导因子而形成外泌体。这些转化系统将允许在植物体内评估预计在植物寄生中起作用的基因。

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