论文部分内容阅读
玉米是重要的粮食作物,水稻黑条矮缩病毒(RBSDV)是玉米粗缩病的病原,由其引起的玉米粗缩病给玉米生产造成重大损失。利用人工mi RNA构建抗病毒植物的技术已经在多种植物中被证明有效,但是在玉米中的尝试未见报道。实验根据玉米zea-mi R159a的前体序列和RBSDV基因组中编码功能蛋白的基因和基因沉默抑制子的序列信息设计引物,构建了用于沉默RBSDV编码基因和基因沉默抑制子的ami RNA(Artificial mi RNA)基因。构建p CAMBIA3301-121-ami RNA植物表达载体,利用农杆菌介导法转化玉米自交系综31(Z31)。对转基因玉米进行分子检测,选择mi RNA表达量高的纯合体株系进行自然发病实验,按0-4的分级标准调查玉米粗缩病的严重度。结果表明,转抗粗缩病毒人工mi RNA载体玉米纯合体株系的抗病表现好于野生型玉米,其中针对基因组6的S6-mi R159转基因玉米抗病情况较好。研究表明利用人工mi RNA技术构建抗病毒病玉米新品种是可行的。
Corn is an important food crop. Rice black-streaked dwarf virus (RBSDV) is the causative agent of maize, and the maize sowing caused by it is a major loss to corn production. Techniques for constructing antiviral plants using artificial mi RNAs have proven effective in a variety of plants, but attempts in maize have not been reported. Primers were designed based on the precursor sequence of zea-mi R159a and the sequence information of gene encoding functional protein and gene silencing suppressor in RBSDV genome to construct ami RNA for silencing RBSDV encoding gene and gene silencing suppressor (Artificial mi RNA) gene. The p CAMBIA3301-121-ami RNA plant expression vector was constructed and transformed into maize inbred line Z31 using Agrobacterium-mediated method. Molecular detection of transgenic maize, selection of homozygous lines with high expression of miRNA for natural pathogenesis experiments, according to 0-4 grading standards to investigate the severity of maize rough. The results showed that the anti-virus performance of the artificial miRNA vector maize homozygous transformed with the anti-cull virus was better than that of the wild-type maize. The resistant condition of S6-mi R159 transgenic maize to genome 6 was better. Studies have shown that the use of artificial mi RNA technology to build a new anti-virus maize varieties is feasible.