Herron C M, Mirkov T E, da Graça J V, Lee R F
Texas A and M University-Kingsville Citrus Center, Weslaco, TX, USA.
J Virol Methods. 2006 Jun;134(1-2):205-11. doi: 10.1016/j.jviromet.2006.01.006. Epub 2006 Feb 21.
Citrus tristeza virus (CTV) is transmitted by several aphid species in a semi-persistent manner with Toxoptera citricida, the brown citrus aphid (BrCA), being the most efficient. As yet, the molecular interactions between the virus and its aphid vectors have not been determined. This is the first report of aphids acquiring CTV from preparations through an artificial membrane and then transmitting it to receptor plants. The BrCA fed across artificial membranes on crude tissue preparations made from CTV-infected bark tissue were able to transmit CTV to virus-free receptor plants at low rates. CTV p20, p27 and p25 proteins, detected by Western blots, were present in all crude tissue preparations from CTV-infected plants. Partially purified CTV preparations were not transmitted by the BrCA in this manner. Infectivity immunoneutralization experiments were conducted where aphids were forced to feed in vitro on three CTV-specific antibodies (p25, p27 and p20) before being placed on receptor plants following a 48h acquisition feed on CTV-infected source plants. There were no differences in transmission rates among the majority of treatments and the control treatments. However, in one infectivity immunoneutralization experiment, the CTV p20 antibodies significantly enhanced CTV transmission compared to buffer only, pre-immune antiserum or no antibody control treatments. This suggests the inactivity of CTV p20 aids BrCA transmission of virions.
柑橘衰退病毒(CTV)可由多种蚜虫以半持久性方式传播,其中褐橘蚜(Toxoptera citricida,BrCA)传播效率最高。目前,病毒与其蚜虫传播介体之间的分子相互作用尚未明确。本文首次报道了蚜虫通过人工膜从制剂中获取CTV,然后将其传播给受体植物。以感染CTV的树皮组织制成的粗组织制剂为食,通过人工膜取食的褐橘蚜能够以较低的速率将CTV传播给无病毒的受体植物。通过蛋白质免疫印迹法检测发现,在所有来自感染CTV植物的粗组织制剂中均存在CTV p20、p27和p25蛋白。部分纯化的CTV制剂不能以这种方式被褐橘蚜传播。进行了感染性免疫中和实验,让蚜虫在以感染CTV的源植物为食48小时后,在体外强迫其取食三种CTV特异性抗体(p25、p27和p20),然后再将它们放置在受体植物上。大多数处理组和对照组之间的传播率没有差异。然而,在一项感染性免疫中和实验中,与仅用缓冲液、免疫前抗血清或无抗体对照组处理相比,CTV p20抗体显著提高了CTV的传播率。这表明CTV p20的失活有助于褐橘蚜传播病毒粒子。