Department of Plant Pathology, University of Nebraska Lincoln, Lincoln, NE 68583, USA.
BMC Plant Biol. 2012 Mar 9;12:33. doi: 10.1186/1471-2229-12-33.
The development of plant gene transfer systems has allowed for the introgression of alien genes into plant genomes for novel disease control strategies, thus providing a mechanism for broadening the genetic resources available to plant breeders. Using the tools of plant genetic engineering, a broad-spectrum antimicrobial gene was tested for resistance against head blight caused by Fusarium graminearum Schwabe, a devastating disease of wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) that reduces both grain yield and quality.
A construct containing a bovine lactoferrin cDNA was used to transform wheat using an Agrobacterium-mediated DNA transfer system to express this antimicrobial protein in transgenic wheat. Transformants were analyzed by Northern and Western blots to determine lactoferrin gene expression levels and were inoculated with the head blight disease fungus F. graminearum. Transgenic wheat showed a significant reduction of disease incidence caused by F. graminearum compared to control wheat plants. The level of resistance in the highly susceptible wheat cultivar Bobwhite was significantly higher in transgenic plants compared to control Bobwhite and two untransformed commercial wheat cultivars, susceptible Wheaton and tolerant ND 2710. Quantification of the expressed lactoferrin protein by ELISA in transgenic wheat indicated a positive correlation between the lactoferrin gene expression levels and the levels of disease resistance.
Introgression of the lactoferrin gene into elite commercial wheat, barley and other susceptible cereals may enhance resistance to F. graminearum.
植物基因转移系统的发展允许将外源基因导入植物基因组，以制定新的疾病控制策略，从而为植物育种者提供了拓宽遗传资源的机制。利用植物遗传工程的工具，对广谱抗菌基因进行了测试，以抵抗由禾谷镰刀菌（Fusarium graminearum Schwabe）引起的穗枯病，这种病是小麦（Triticum aestivum L.）和大麦（Hordeum vulgare L.）的毁灭性疾病，会降低谷物的产量和质量。
使用含有牛乳铁蛋白 cDNA 的构建体，通过农杆菌介导的 DNA 转移系统转化小麦，以在转基因小麦中表达这种抗菌蛋白。通过 Northern 和 Western blot 分析转化体，以确定乳铁蛋白基因的表达水平，并接种穗枯病真菌 F. graminearum。与对照小麦植株相比，转基因小麦表现出由 F. graminearum 引起的病害发生率显著降低。与对照 Bobwhite 以及两个未转化的商业小麦品种易感 Wheaton 和耐 ND 2710 相比，高度易感的 Bobwhite 小麦品种中，转基因植株的抗性水平显著更高。通过 ELISA 对转基因小麦中表达的乳铁蛋白蛋白进行定量，表明乳铁蛋白基因表达水平与疾病抗性水平之间存在正相关。
将乳铁蛋白基因导入优良的商业小麦、大麦和其他易感谷物中，可能会增强对禾谷镰刀菌的抗性。
