基于RNAi抗PRSV番木瓜株系的培育及其分子特征验证  

Breeding and Molecular Characteristion of RNAi Papaya for Resistance of Papaya Ringspot Virus

作　　者：黄静[1,2,3] 贾瑞宗[2] 孔华[2] 郭静远[2] 王绪朋 郭安平[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所,海南海口571101 [3]海南医学院基础医学与生命科学学院,海南海口571199

出　　处：《热带作物学报》2017年第3期513-520,共8页Chinese Journal of Tropical Crops

基　　金：农业部948项目(No.2014-Z17;No.2015-Z23);海南省重点项目(No.ZDYF2016027);海南省自然科学基金(No.20153145)

摘　　要：根据RNAi原理构建的抗海南番木瓜环斑病毒(PRSV)植物表达载体p CAMBIA2300-35S-CP-RNAi-OCS,通过基因枪法将载体导入番木瓜的愈伤组织中,经卡那霉素抗性筛选后再分化成为植株。描述转化植株的分子特征同时进行抗病毒试验分析其抗病性。实验结果显示:转基因株系474为单拷贝插入的杂合子,插入位点在第7号染色体supercontig_61的717 141位置;抗病毒试验中,在接种病毒后28 d内非转基因株系1280有高浓度的病毒积累并很快表现出明显的病症,而转基因株系474基本无病毒积累且无发病症状。表明转基因株系474具有较好的PRSV抗性,有待于进一步田间试验。RNAi-mediated virus resistance strategy was used to develop PRSV resistance papaya lines. A plant expression vector p CAMBIA2300-35S-CP-RNAi-OCS was constructed. The vector was transferred into papaya callus by particle bombardment, selected on kanamycin media. Transformation papaya lines were validated by PCR, RTPCR, dd PCR, etc. The transgenic lines were further challenged with viral inoculation. The results showed that transgenic line 474 was single copy insertion and insertion site was located in papaya chromosome 7 supercontig_61at 717 141 bp; non-transgenic line 1280 accumulated higher concentration of virus and developed symptoms, while transgenic line 474 was almost no virus accumulation. Transgenic line 474 was resistance to PRSV, which needed field test.

关 键 词：番木瓜环斑病毒 番木瓜 RNAI 

分 类 号：S188[农业科学—农业基础科学]