Osusky M, Zhou G, Osuska L, Hancock R E, Kay W W, Misra S
Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 3P6, Canada.
Nat Biotechnol. 2000 Nov;18(11):1162-6. doi: 10.1038/81145.
Here we describe a strategy for engineering transgenic plants with broad-spectrum resistance to bacterial and fungal phytopathogens. We expressed a synthetic gene encoding a N terminus-modified, cecropin-melittin cationic peptide chimera (MsrA1), with broad-spectrum antimicrobial activity. The synthetic gene was introduced into two potato (Solanum tuberosum L.) cultivars, Desiree and Russet Burbank, stable incorporation was confirmed by PCR and DNA sequencing, and expression confirmed by reverse transcription (RT)-PCR and recovery of the biologically active peptide. The morphology and yield of transgenic Desiree plants and tubers was unaffected. Highly stringent challenges with bacterial or fungal phytopathogens demonstrated powerful resistance. Tubers retained their resistance to infectious challenge for more than a year, and did not appear to be harmful when fed to mice. Expression of msrA1 in the cultivar Russet Burbank caused a striking lesion-mimic phenotype during leaf and tuber development, indicating its utility may be cultivar specific. Given the ubiquity of antimicrobial cationic peptides as well as their inherent capacity for recombinant and combinatorial variants, this approach may potentially be used to engineer a range of disease-resistant plants.
在此,我们描述了一种培育对细菌和真菌植物病原体具有广谱抗性的转基因植物的策略。我们表达了一个合成基因,该基因编码一种N端修饰的天蚕素-蜂毒素阳离子肽嵌合体(MsrA1),具有广谱抗菌活性。将该合成基因导入两个马铃薯(Solanum tuberosum L.)品种德西蕾(Desiree)和褐皮伯班克(Russet Burbank),通过PCR和DNA测序确认了其稳定整合,并通过逆转录(RT)-PCR和生物活性肽的回收确认了表达。转基因德西蕾植株和块茎的形态及产量未受影响。用细菌或真菌植物病原体进行的高度严格挑战显示出强大的抗性。块茎对感染性挑战的抗性保持了一年多,喂食小鼠时似乎也没有危害。MsrA1在褐皮伯班克品种中的表达在叶片和块茎发育过程中引起了显著的类病斑表型,表明其效用可能具有品种特异性。鉴于抗菌阳离子肽的普遍性及其重组和组合变体的固有能力,这种方法可能潜在地用于培育一系列抗病植物。