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小麦纹枯病(主要病原为禾谷丝核菌)和赤霉病(主要病原为禾谷镰刀菌)已成为我国小麦生产的重要病害。TaPIEP1是从小麦中分离到的1个病原诱导的基因,其编码蛋白是可与GCC-box顺式元件结合、转录激活型的ERF转录因子。本研究以8个转TaPIEP1基因小麦株系的T4和T5代植株为试材,进行了外源转TaPIEP1基因的PCR检测、Southern杂交、RT-PCR与Q-RT-PCR的分析以及纹枯病菌、赤霉病菌接种与抗性鉴定。结果表明,外源TaPIEP1基因在转基因小麦中能够稳定遗传,以单拷贝或双拷贝整合到7个转基因小麦株系基因组的不同位点;外源TaPIEP1基因在转基因小麦中能超量表达;与受体扬麦12相比,TaPIEP1表达水平高的8个转基因小麦株系对纹枯病抗性显著提高,4个株系中一些材料对赤霉病抗性显著提高,3个株系中一些材料兼抗纹枯病和赤霉病,说明TaPIEP1正向参与了小麦对纹枯病和赤霉病抗性反应,利用该基因通过基因工程可创制抗纹枯病、赤霉病的小麦新种质。
Wheat sheath blight (the main pathogen is Rhizoctonia cerealis) and scab (the main pathogen is Fusarium graminear) has become an important disease of wheat production in our country. TaPIEP1 is a pathogen-induced gene isolated from wheat and encodes an ERF transcription factor that binds to the GCC-box cis-element and is transcriptionally active. In this study, T4 and T5 plants from eight TaPIEP1 transgenic wheat lines were used as materials for PCR analysis of TaPIEP1 gene. Southern hybridization, RT-PCR and Q-RT-PCR analysis and sheath blight Fungus, Fusarium graminearum inoculation and resistance identification. The results showed that the exogenous TaPIEP1 gene can be stably inherited in transgenic wheat and integrated into different genomes of seven transgenic wheat lines in one or two copies. The exogenous TaPIEP1 gene can be overexpressed in transgenic wheat, Eight transgenic wheat lines with higher expression levels of TaPIEP1 showed significantly higher resistance to sheath blight than those with positive ones. Some of the four lines had significantly increased resistance to scab, and some of the three lines And resistance to sheath blight and scab, indicating that TaPIEP1 is positively involved in the resistance of wheat to sheath blight and scab. By utilizing this gene, genetic engineering can create new wheat germplasms resistant to scab, .