Separation of Citrus Tristeza Virus (CTV) Serotypes Through Aphid Transmission
Separation of Citrus Tristeza Virus (CTV) Serotypes Through Aphid Transmission
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通过蚜虫传播分离柑橘 Tristeza 病毒 (CTV) 血清型
DOI：
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
M. Koizumi
中科院分区：
文献类型：
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作者：
T. Kano;M. Koizumi
通过柑橘弓形虫（Toxoptera citricidus, Kirk）的传播，从15个不同的原野源获得了柑橘tristeza病毒（CTV）的20个不同的传代培养物。采用单克隆抗体（MAb） MCA13和3DF1和多克隆抗体（PAb）双夹心ELISA对20份蚜虫传代培养（ATS）和15份田间源进行检测。从两个田间源（FS）分离得到的4株ATS表现出与其亲本FS不同的反应。例如，来自一个FS M27的温和CTV对MCA13、3DF1和PAb反应强烈，而来自M27的三个ATS分离株与PAb反应，但对两种MAb都不反应。这些结果表明，至少两种不同的血清型可以感染一株柑橘植株，并且某些血清型可以通过蚜虫传播而分离。关键词：ELISA，单克隆抗体。在相应的论文(6)中，利用单克隆抗体（MAb）鉴定了来自日本不同产地（FS）的柑橘tristeza病毒（CTV）的血清学多样性，这些单克隆抗体对CTV外壳蛋白的不同表位具有特异性(3)。已知CTV在野外以不同分离物的复合体或混合物的形式存在，一些工作者已经报道了从这些复合体中分离出组分。例如，通过对受感染植株进行热处理后的组织嫁接(I)、媒介传播(9)和从幼苗发黄后恢复的树木（SY）进行组织嫁接（14），可以分离出温和菌株。本文比较了亲本F S培养物和蚜虫传代培养物（ATS）与MCA13（10）和3DF1（13）两种单克隆抗体和一种多克隆抗体（PAb）的反应，以确定它们在蚜虫传代后的血清学反应是否发生变化。数据表明，蚜虫可以从同一植物的混合物中分离出特定的CTV成分，并通过使用差异单抗进行检测。材料和方法。所测试的CTV fs如表1所示，之前已经描述过（4,5,6,7）。蚜虫向量。采自果树研究站（FTRS） Okitsu分站的柑橘林，用人工饲料(8)饲养至少8天，获得一个无毒蚜虫菌落。每2天更换一次饮食（包装在玻璃杯中实验室薄膜袋内）。将蚜虫转移到对CTV具有抗性的健康三叶橙幼苗上。病毒的传播。供体植株分别接枝粗柠檬或葡萄夫人甜橙幼苗，或F S品种接枝粗柠檬幼苗。在20-25℃的空调室内，蚜虫在获取宿主的新花枝上饲养至少7天。在3-6株受体植株上分别移植20多只毒蚜。受体植株为6 ~ 12个月大的健康葡萄夫人甜橙或粗柠檬幼苗。在20-25℃的空调室中，初始饲养周期为48小时或更长。每次接种后。在植物上喷洒杀虫剂以消灭病媒。病毒的来源。病毒源酶联免疫吸附植物和所有其他植物进行实验（ELISA）。在无载体夹心中保持双抗体（@AS），采用酶联免疫吸附试验（ELISA）和多分子抗体（PAb）对贡献果树Res. Sta进行验证。b - 168。受体植物的CTV感染2
Twenty different subcultures of citrus tristeza virus (CTV) were obtained from 15 different original field sources by transmission with the aphid Toxoptera citricidus (Kirk). The 20 aphid-transmitted subcultures (ATS), and 15 field sources were tested by double sandwich indiret ELISA using two monoclonal antibodies (MAb), MCA13 and 3DF1, and by double sandwich ELISA using polyclonal antibodies (PAb). Four ATS isolates derived from two field source (FS) isolates showed reactions different from their parent FS. For example, a mild CTV from one FS, M27, reacted strongly to MCA13, 3DF1 and PAb, while three ATS isolates from M27 reacted with PAb, but not with either MAb. These results indicate that a t least two different serotypes can infect one citrus plant, and that separation of certain serotypes can occur through aphid transmission. Index words: ELISA, monoclonal antibody. In the accompanying paper (6), the serological diversity of citrus tristeza virus (CTV) from different field sources (FS) in Japan was identified using monoclonal antibodies (MAb) which are specific to different epitopes in the CTV coat protein (3). CTV is known to exist as a complex or mixture of different isolates in the field, and separation of components from these complexes has been reported by several workers. For example, separation of mild strains has been accomplished by tissue grafting after heat treatment of infeced plants (I), by vector transmission (9), and by tissue grafting from trees which showed recovery from seedling yellows (SY) (14). In this paper, we compared the reactions of parent F S cultures and aphid-transmitted subcultures (ATS) with two MAbs, MCA13 (10) and 3DF1 (13)) and a polyclonal antibody (PAb) to determine whether their serological reactions change after aphid transmission. Data are presented indicating that separation of a specific CTV component from a mixture of components in the same plant can be achieved by aphids and detected by use of differential MAb. MATERIALS AND METHODS screenhouses. The CTV F S tested are shown in Table 1 and have been previously described (4, 5, 6, 7). Aphid vectors. Toxoptera citricidus (Kirk.) was collected from a citrus grove a t the Okitsu Branch, Fruit Tree Research Station (FTRS) and reared on synthetic diet (8) for at least 8 days to obtain a colony of nonviruliferous aphids. The diet (enclosed in laboratory film sachets on a glass cup) was changed every 2 days. The aphids were transferred to healthy trifoliate orange seedlings, which are resistant to CTV. Virus transmission. Donor plants were graft-inoculated rough lemon or Madam Vinous sweet orange seedlings or the F S cultivar grafted to rough lemon seedlings. Aphids were reared on new flushes of the acquisition host for at least 7 days at 20-25 C in an air-conditioned chamber. More than 20 viruliferous aphids were transferred to each of 3-6 receptor plants. The receptor plants were healthy Madam Vinous sweet orange or rough lemon seedlings 6 to 12 months old. The inouclation feeding period was 48 hr or more at 20-25 C in an air-conditioned chamber. After each inoculation. plants were sprayed with insecticide to eliminate the vectors. Virus sources. The virus source Enzyme-linked immunosorbent plants and all other plants for experiassay (ELISA). Double antibody ments were maintained in vector-free sandwich @AS) ELISA with polyelonal antibody (PAb) was used to verify Contribution Fruit Trees Res. Sta. B-168. CTV infection in the receptor plants 2
针对 GPR40 的单克隆抗体的生产和表征。
DOI：
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发表时间：
2008
期刊：
Biochem Biophys Res Commun 365
影响因子：
--
作者：
Hirasawa;A
通讯作者：
A