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Restoring pollen fertility in transgenic male-sterile eggplant by Cre/loxp-mediated site-specific recombination system

机译:通过Cre / loxp介导的位点特异性重组系统恢复转基因雄性不育茄子的花粉育性

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摘要

This study was designed to control plant fertility by cell lethal gene Barnase expressing at specific developmental stage and in specific tissue of male organ under the control of Cre/loxP system, for heterosis breeding, producing hybrid seed of eggplant. The Barnase-coding region was flanked by loxP recognition sites for Cre-recombinase. The eggplant inbred/pure line (‘E-38') was transformed with Cre gene and the inbred/pure line (‘E-8') was transformed with the Barnase gene situated between loxp. The experiments were done separately, by means of Agrobacterium co-culture. Four T0 -plants with the Barnase gene were obtained, all proved to be male-sterile and incapable of producing viable pollen. Flowers stamens were shorter, but the vegetative phenotype was similar to wild-type. Five T 0 -plants with the Cre gene developed well, blossomed out and set fruit normally. The crossing of male-sterile Barnase-plants with Cre expression transgenic eggplants resulted in site-specific excision with the male-sterile plants producing normal fruits. With the Barnase was excised, pollen fertility was fully restored in the hybrids. The phenotype of these restored plants was the same as that of the wild-type. Thus, the Barnase and Cre genes were capable of stable inheritance and expression in progenies of transgenic plants.
机译:本研究旨在通过在Cre / loxP系统的控制下在特定发育阶段和男性器官的特定组织中表达的细胞致死基因Barnase控制植物的育性,进行杂种育种,生产茄子的杂种。 Barnase编码区的侧翼是Cre重组酶的loxP识别位点。用Cre基因转化茄子近交/纯品系('E-38'),并通过位于loxp之间的Barnase基因转化近交/纯品系('E-8')。通过农杆菌共培养分别进行实验。获得了具有Barnase基因的四株T0植物,全部被证明是雄性不育的并且不能产生活的花粉。花雄蕊较短,但营养表型与野生型相似。五株带有Cre基因的T 0植株发育良好,开花并正常结实。雄性不育的Barnase植物与Cre表达的转基因茄子的杂交导致了定点切除,而雄性不育的植物产生了正常的果实。切下Barnase后,杂种中的花粉育性得以完全恢复。这些恢复的植物的表型与野生型相同。因此,Barnase和Cre基因能够在转基因植物的子代中稳定地遗传和表达。

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