Meiyalaghan Sathiyamoorthy, Mohan Sara, Pringle Julie M, Baldwin Samantha, Jacobs Jeanne M E, Conner Anthony J
New Zealand Institute for Plant & Food Research Limited, Private Bag 4704, Christchurch, New Zealand.
Commun Agric Appl Biol Sci. 2009;74(3):667-79.
Disease resistance is an important objective of global potato breeding programmes. The use of resistant cultivars is a significant tool for disease management. Recent advances in plant molecular genetics have identified several genes for resistance to potato diseases from within the germplasm pool available to potato breeders. Antimicrobial peptides, such as Snakin-1 (StSN1) and Snakin-2 (StSN2), have been isolated recently from potato tubers. Overexpression of the StSNI and StSN2 genes in potato is known to provide broad spectrum activity against a wide range of bacterial and fungal pathogens. We describe the use of intragenic gene transfer technology towards disease resistance in potatoes. An expression cassette was constructed with the 5' promoter and 3' terminator regions of a potato gene encoding a chlorophyll a/b binding protein (StLhca3). The coding regions of the StSN1 and StSN2 genes of potato were cloned individually between these regulatory regions. The resulting Lhca3-StSNi-Lhca3 and Lhca3-StSN2-Lhco3 chimeric genes were individually cloned into a potato-derived T-DNA-like region for potato transformation. Potato cultivar Iwa was co-cultivated with Agrobocterium harbouring intragenic binary vectors with the StSN1 and StSN2 genes. Regenerated potato plants were screened using PCR to identify lines transformed with the disease resistance genes without the presence of foreign DNA.
抗病性是全球马铃薯育种计划的一个重要目标。使用抗病品种是病害管理的一项重要手段。植物分子遗传学的最新进展已从马铃薯育种者可用的种质库中鉴定出几个抗马铃薯病害的基因。抗菌肽,如蛇形蛋白-1(StSN1)和蛇形蛋白-2(StSN2),最近已从马铃薯块茎中分离出来。已知在马铃薯中过表达StSN1和StSN2基因可提供针对多种细菌和真菌病原体的广谱活性。我们描述了利用基因内基因转移技术实现马铃薯抗病性的方法。构建了一个表达盒,其中包含编码叶绿素a/b结合蛋白(StLhca3)的马铃薯基因的5'启动子和3'终止子区域。马铃薯的StSN1和StSN2基因的编码区分别克隆在这些调控区域之间。将得到的Lhca3-StSN1-Lhca3和Lhca3-StSN2-Lhca3嵌合基因分别克隆到一个源自马铃薯的类T-DNA区域用于马铃薯转化。将马铃薯品种Iwa与携带含有StSN1和StSN2基因的基因内二元载体的农杆菌共培养。使用PCR筛选再生的马铃薯植株,以鉴定在不存在外源DNA的情况下用抗病基因转化的株系。