摘要
康乃馨(Dianthus caryophyllus L.)是全球贸易量最大的鲜切花之一。转基因康乃馨因颜色新奇、花型丰富、插瓶期长等特点深受消费者喜爱。现有19个转基因康乃馨品系列入全球商业化种植,其中18个品系因转入了类黄酮-3’,5’-羟化酶(flavonoid-3’,5’-hydroxylase,F3’5’H)基因而产生了蓝紫色花瓣的性状。18个品系转入的F3’5’H基因中11个来源于大花三色堇(bp40基因),还有7个来源于矮牵牛(hf1基因)。我国目前没有批准转基因康乃馨进口,更缺乏相关检测方法。建立转基因康乃馨外源基因筛查方法,用以防范转基因康乃馨种苗、鲜切花等非法进入我国显得至关重要。本研究针对转基因康乃馨中最常见的两种不同来源F3’5’H基因,建立了普通PCR和实时荧光PCR检测方法,构建了适于两个基因检测的质粒分子,并将这两种方法应用于进境产品检测。结果表明,建立的bp40基因和hf1基因检测方法均具有特异性,普通PCR方法的检测下限(LOD)可分别达到25拷贝和12.5拷贝,实时荧光PCR方法的LOD均为10拷贝,在进境康乃馨种苗等样品中未检出该基因。综上,本研究建立的两种F3’5’H基因检测方法适用于康乃馨及其来源产品的转基因筛查检测,为防止未经我国批准的转基因产品非法进入我国提供有效的技术支撑。
Carnation(Dianthus caryophyllus L.)is one of the most traded fresh cut flowers globally.GM carnations are popular among consumers because of their novel colors,rich flower patterns,and long vase life.There are currently 19 GM carnation events listed in commercial cultivation worldwide,of which 18 events have been transferred to the flavonoid-3',5'-hydroxylase gene(F3'5'H)endows the trait of blue-purple petals.Among the 18 events,11 F3'5'H genes were derived from Pansy grandiflora(bp40 gene),and 7 were derived from petunia(hfl gene).Our country has not yet approved the import of GM carnations,and lacks relevant detection methods.Therefore,in order to prevent GM carnation seedlings and fresh cut flowers from illegally entering China,it is very important to establish screening methods for exogenous genes in GM carnations.In this study,PCR and real-time PCR detection methods for two F3'5'H genes were established,and plasmid molecules suitable for the detection of the two genes were construct-ed,and the method was applied to the imported products detection.Results showed that the established bp40 and hfl gene detection methods are specific to the relative gene detection,the limit of detection(LOD)of the PCR method can reach 25 copies and 12.5 copies,respectively,and the LOD of the re-al-time PCR method can reach 10 copies.The bp40 and hfl genes could not be detected in imported carnation seedlings and other products.In summary,the two F3'5'H gene detection methods established in this study are suitable for screening of GM carnations and their derived products.These methods provided the effective technical means to prevent the illegal entry of GM products not approved by China.
作者
钱昌元
尹璐
傅怡宁
左翠花
李想
Qian Changyuan;Yin Lu;Fu Yining;Zuo Cuihua;Li Xiang(Animal,Plant and Inspection and Quarantine Center,Shanghai Customs District,Shanghai 200135,China)
出处
《植物检疫》
2023年第6期45-51,共7页
Plant Quarantine
基金
上海市“科技创新行动计划”技术标准项目(21DZ2203000)
海关总署科研项目(2021HK155)。
