甜菜坏死黄脉病毒（BNYVV）的快速检测技术及在抗病育种上的应用  

Idirect-ELISA and Its Utilization on Breeding Sugarbeet Resistance to Rhizomania

作　　者：张春来[1] 孙以楚[1] 王华忠[1] 魏培文[1] 张明厚[1] 

机构地区：[1]中国农业科学院甜菜研究所,东北农业大学

出　　处：《中国甜菜》1995年第2期21-24,共4页

基　　金：中国农业科学院院长基金

摘　　要：应用ＢＮＹＶＶ抗体球蛋白，建立了快速检测ＢＮＹＶＶ的间接酶跌免疫吸附技术（Ｉｎｄｉｒｅｃｔ－ＥＬＬＳＡ），该法检测含抗原材料仅需４．５小时，较国际通用程序缩短２０余小时。主要程序包括：抗原经３７℃包被２小时后，加ＢＮＹＶＶ抗体球蛋白ＩｇＧ，３７℃孵育１小时，然后加过氧化物酶标记的单抗兔ＩｇＧ，３７℃孵育１小时，最后加底物液邻笨二胺，常温下反应１５～３０分钟，应用该法对采集的４份标样进行测定，其中两份测出含有ＢＮＲＶＶ，对不同抗感丛根病性品种（系）经室内病土盆栽或病田栽培，调查病情并取样测定根部ＢＮＹＶＶ含量，结果表明。品种间存在显著差异，ＢＮＹＶＶ含量、病情与品种（系）抗病性表现基本一致。A rapid indirect ELISA detecting technique to BNYVV has been established. only need4.5hrs to detect samples contained BNYVV,using this technique and 20hrs shorten comparedwith the current methods in the world,The main proceduce include:the polystyrene walls ofthe microtibe plates are first coated with antigen or samples,incubated for 2hrs at 37℃.Add BNYVV antibody gfobulin IgG to the absorbed antigen,incubatal for l hr at 37℃.Add horoe radish peroxidase(HRR)-conjugated anti-rabbit IgG in goats, incubate for lhr at37℃, Add freshly prepared substrate 0-phenylenediamine and leave at room temperaturefor colour to develope,Two of four samples from fields of our Institute detected Rhizomaniainfested.Lines or varieties with different level of resistance to Rhizomania cultured in infestedsoil of pot and infested fields.Investigation of disease sympto ms and determination ofBNYVV concentration of samples confirmed significant difference among lines or cultivars.BNYVV concentration,symptoms were almost coi responding to disease resistance of lines orailtivars.

关 键 词：甜菜 丛根病 抗病育种 

分 类 号：S566.303.4[农业科学—作物学]