The National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Crop Genetic and Breeding of the Agriculture Ministry, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China.
Funct Integr Genomics. 2012 Aug;12(3):481-8. doi: 10.1007/s10142-012-0286-z. Epub 2012 Jun 12.
The fungus Cochliobolus sativus is the main pathogen of common root rot, a serious soil-borne disease of wheat (Triticum aestivum L.). The fungus Fusarium graminearum is the primary pathogen of Fusarium head blight, a devastating disease of wheat worldwide. In this study, the wheat lipid transfer protein gene, TaLTP5, was cloned and evaluated for its ability to suppress disease development in transgenic wheat. TaLTP5 expression was induced after C. sativus infection. The TaLTP5 expression vector, pA25-TaLTP5, was constructed and bombarded into Chinese wheat variety Yangmai 18. Six TaLTP5 transgenic wheat lines were established and characterized. PCR and Southern blot analyses indicated that the introduced TaLTP5 gene was integrated into the genomes of six transgenic wheat lines by distinct patterns, and heritable. RT-PCR and real-time quantitative RT-PCR revealed that the TaLTP5 gene was over-expressed in the transgenic wheat lines compared to segregants lacking the transgene and wild-type wheat plants. Following challenge with C. sativus or F. graminearum, all six transgenic lines overexpressing TaLTP5 exhibited significantly enhanced resistance to both common root rot and Fusarium head blight compared to the untransformed wheat Yangmai 18.
禾旋孢腔菌是小麦普通根腐病（一种严重的土传小麦病害）的主要病原菌，而禾谷镰刀菌则是小麦赤霉病（一种全球范围内破坏性极大的小麦病害）的主要病原菌。在本研究中，克隆了小麦脂质转移蛋白基因 TaLTP5，并评估了其在转基因小麦中抑制病害发展的能力。TaLTP5 在受到禾旋孢腔菌感染后表达上调。构建了 TaLTP5 表达载体 pA25-TaLTP5，并将其轰击到中国小麦品种扬麦 18 中。建立并鉴定了 6 个 TaLTP5 转基因小麦株系。PCR 和 Southern blot 分析表明，引入的 TaLTP5 基因通过不同的模式整合到 6 个转基因小麦株系的基因组中，并且可遗传。RT-PCR 和实时定量 RT-PCR 显示，与不含转基因的分离株和野生型小麦植株相比，转基因小麦株系中 TaLTP5 基因过表达。在受到禾旋孢腔菌或禾谷镰刀菌的攻击后，所有过表达 TaLTP5 的 6 个转基因株系均表现出对普通根腐病和小麦赤霉病的显著增强抗性，而未转化的小麦扬麦 18 则表现出较弱的抗性。
