摘要
槟榔是海南重要的经济作物,APV1 (areca palm velarivirus 1)病毒引起的槟榔黄化病严重危害槟榔产业的健康发展。研发快速精准的APV1病毒检测技术对于防控槟榔黄化病具有重要意义。本研究利用RT-PCR扩增APV1病毒的外壳蛋白(CP)基因序列,构建原核表达载体pET30a-APV1-CP并转化至大肠杆菌,经诱导表达和蛋白纯化获得APV1-CP蛋白。通过注射兔子获得多克隆抗体,经检测其效价为102 400。采用ELISA和RT-PCR分别对槟榔APV1病毒检测,发现两种检测方法大部分的检测结果基本一致。APV1病毒ELISA检测技术对槟榔种苗的检测及槟榔黄化病的防控具有重要的实用价值。
Betel palm is an important tropical crop in Hainan. APV1(areca palm velari virus 1) virus caused betel nut yellowing disease seriously endangered the healthy development of the betel nut industry. Research and development of fast and accurate APV1 virus detection technology is important for disease prevention and control of yellow betel. In this study, RT-PCR was used to amplify the coat protein(CP) gene sequence of the APV1 virus,and the prokaryotic expression vector pET30 a-APV1-CP was constructed and transformed into E. coli. The APV1-CP protein was obtained by induced expression and protein purification. The polyclonal antibody was obtained by injection of APV1-CP into rabbit, and its titer was identified as 102 400. Subsequently, ELISA and RT-PCR were used to detect APV1 virus in betel palm, and it was found that most of the results of the two methods were consistent. APV1 virus ELISA detection technology has important practical value for the detection of betel nut seedlings and the prevention and control of betel palm YLD.
作者
陈阳
王洪星
黄惜
Chen Yang;Wang Hongxing;Huang Xi(Key Laboratory of Sustainable Utilization of Tropical Biological Resources of Hainan Province,School of Tropical Crops,Hainan University,Haikou,570228)
出处
《分子植物育种》
CAS
北大核心
2022年第2期518-523,共6页
Molecular Plant Breeding
基金
海南省重大科技计划项目(ZDKJ201817)资助。
