Leclerc D, AbouHaidar M G
Department of Botany, University of Toronto, Ontario, Canada.
Mol Plant Microbe Interact. 1995 Jan-Feb;8(1):58-65. doi: 10.1094/mpmi-8-0058.
Four constructs of the potato aucuba mosaic virus (PAMV) coat protein (CP) gene were engineered for expression in tobacco plants: The full-length CP sequence (FL CP), two truncated forms--one at the N terminus (46 amino acid residues are deleted) (NT138), one in the conserved core portion (86 amino acids deleted) (CT258) of the gene--and an antisense RNA construct. These constructs were introduced into tobacco plants (Nicotiana tabacum) via Agrobacterium tumefaciens-mediated transformation. The plants transformed with the NT138 and FL CP constructs produced the mRNAs and proteins from the respective transgene. Transformants with the CT258 construct produced the transgenic mRNA, but the modified CP was not detected in the 20 different transformants tested. Transgenic R0 and R1 tobacco plants expressing the full-length, CT258, and the antisense constructs exhibited protection to PAMV infection and a delay in symptom development when inoculated with 0.1 and 0.5 microgram/ml of purified PAMV. Transgenic plants expressing the NT138 construct did not confer any detectable protection to PAMV infection. These results suggest that an engineered coat protein mediated resistance (CPMR) can be obtained from a CP gene truncated in its core region. The role of the N-terminal domain of the CP in the CPMR of PAMV and the implication of either the RNA or the protein in the protection is discussed.
构建了马铃薯奥古巴花叶病毒(PAMV)外壳蛋白(CP)基因的四种构建体,用于在烟草植株中表达:全长CP序列(FL CP)、两种截短形式——一种在基因的N端(缺失46个氨基酸残基)(NT138),一种在基因的保守核心部分(缺失86个氨基酸)(CT258)——以及一个反义RNA构建体。通过根癌农杆菌介导的转化将这些构建体导入烟草植株(烟草)。用NT138和FL CP构建体转化的植株产生了来自各自转基因的mRNA和蛋白质。用CT258构建体转化的植株产生了转基因mRNA,但在测试的20个不同转化体中未检测到修饰的CP。表达全长、CT258和反义构建体的转基因R0和R1烟草植株在接种0.1和0.5微克/毫升纯化的PAMV时,表现出对PAMV感染的抗性以及症状发展延迟。表达NT138构建体的转基因植株对PAMV感染未提供任何可检测到的抗性。这些结果表明,可以从其核心区域截短的CP基因获得工程化外壳蛋白介导的抗性(CPMR)。讨论了CP的N端结构域在PAMV的CPMR中的作用以及RNA或蛋白质在抗性中的意义。