• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Turkish Journal of Agriculture & Forestry >Genetic transformation of Citrus paradisi with antisense and untranslatable RNA-dependent RNA polymerase genes of Citrus tristeza closterovirus.
【24h】

Genetic transformation of Citrus paradisi with antisense and untranslatable RNA-dependent RNA polymerase genes of Citrus tristeza closterovirus.

机译：用柑桔丝状体梭状病毒的反义和不可翻译的依赖RNA的RNA聚合酶基因对天堂柑橘进行遗传转化。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Protein and RNA-mediated forms of pathogen-derived resistance (PDR) have been developed against many viruses in different plants. However, no resistance has been reported against Citrus tristeza virus (CTV), a closterovirus, in Citrus species transformed with coat protein genes or other sequences of CTV. The successful use of replication-associated genes in RNA-mediated resistance in other crops prompted the use of the RNA-dependent RNA polymerase (RdRp) gene of CTV for the development of RNA-mediated PDR in Citrus. The RdRP gene was amplified from CTV isolate DPI3800 from Florida and used to generate antisense (RdRp-AS) and untranslatable (RdRp-UT) constructs with point mutation consecutive stop codons in the 5' end of the RdRp gene for use in plant transformation. A total of 3120 etiolated epicotyl segments of Duncan grapefruit (Citrus paradisi Macf. cv. Duncan) were transformed with these constructs using Agrobacterium tumefaciens-mediated transformation. From these segments 1040 kanamycin-resistant shoots were regenerated, and a total of 131 putative transgenic shoots were identified by fluorescent microscopy and histochemical beta -glucuronidase (GUS) assays. One hundred GUS positive plants were rooted and 66 plants survived and were established on soil. A total of 41 plants were tested by polymerase chain reaction (PCR) for the presence of the GUS gene and for the transgenes. Eighteen GUS-positive and transgene-positive plants (8 with RdRp-AS, and 10 with RdRp-UT) were identified..
机译：已经开发出蛋白质和RNA介导的病原体衍生抗性（PDR）形式，可以抵抗不同植物中的许多病毒。但是，在用外壳蛋白基因或其他CTV序列转化的柑橘属物种中，尚无抗梭状芽胞病毒柑橘（CTV）的报道。复制相关基因在其他农作物的RNA介导的抗性中的成功使用，促使CTV的RNA依赖性RNA聚合酶（RdRp）基因用于柑橘中RNA介导的PDR的开发。从佛罗里达州的CTV分离株DPI3800中扩增了RdRP基因，并用于生成反义（RdRp-AS）和不可翻译（RdRp-UT）构建体，该构建体在RdRp基因的5'端具有点突变连续终止密码子，用于植物转化。使用根癌农杆菌介导的转化，用这些构建体转化邓肯葡萄柚（Citrus paradisi Macf.cv.Duncan）的总共3120个黄化的上胚轴片段。从这些片段中再生出1040株抗卡那霉素的芽，并通过荧光显微镜和组织化学β-葡萄糖醛酸苷酶（GUS）分析鉴定出总共131个推定的转基因芽。 GUS阳性植物有一百株，其中66株存活并建立在土壤上。通过聚合酶链反应（PCR）测试了总共41株植物中GUS基因的存在和转基因。鉴定出18种GUS阳性和转基因阳性植物（其中8种具有RdRp-AS，10种具有RdRp-UT）。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号