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首页> 外文期刊>Cytology and genetics >Obtaining Wheat (Triticum aestivum L.) Lines with Yeast Genes for Trehalose Biosynthesis
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Obtaining Wheat (Triticum aestivum L.) Lines with Yeast Genes for Trehalose Biosynthesis

机译：用酵母基因获取小麦（Triticum aestivum L.）线用于海藻糖生物合成

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摘要

Yeast (Saccharomyces cerevisiae) genes for trehalose biosynthesis (TPS1 andTPS2) were transferred into genomes of several common wheat cultivars using two methods ofAgrobacterium-mediated transformation (in vitro and in planta) to enhance drought tolerance. For this purpose, vectors pBract214-TPS1 and pBract214-TPS2 were constructed using the Gateway-cloning technique. Both the vectors containedTPS1andTPS2genes under control of the constitutive maize ubiquitin promoter (PUbi) and selectable marker hygromycin-phosphotransferase (hpt) gene. Three- to five-day-old calluses obtained from immature wheat embryos were used as explants for the transformation in vitro. Selection of transgenic plants was carried out on nutrient medium supplemented with 30 mg/L hygromycin (as a selectable agent). Seeds of wheat (transgenic generation T1) were obtained by the in planta method of transformation. Integration and the presence of yeast genes in wheat genomic DNA isolated from transgenic plants were confirmed by PCR analysis using primers specific toTPS1andTPS2genes.
机译：酵母（酿酒酵母）用于海藻糖生物合成的基因（TPS1和TPS2）使用两种经过委托介导的转化（体外和植物）的方法转移到几种常见小麦品种的基因组中，以增强耐旱性。为此目的，使用网关克隆技术构建载体PBRACT214-TPS1和PBRACT214-TPS2。载体的载体含有在组成型玉米泛素启动子（Pubi）和选择标记潮霉素 - 磷酸转移酶（HPT）基因的控制下进行的。从未成熟的小麦胚胎获得的三天历史的愈伤组织用作体外转化的外植体。在补充有30mg / L潮霉素的营养培养基上进行转基因植物（作为选择剂）。小麦种子（转基因生成T1）通过植物的转化方法获得。通过PCR分析使用引物特异性Totps1Andtps2GeNES通过PCR分析证实了从转基因植物中分离的小麦基因组DNA中的整合和酵母基因的存在。

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