Reinbold C, Gildow F E, Herrbach E, Ziegler-Graff V, Gonçalves M C, van den Heuvel J F J M, Brault V
INRA, 28 rue de Herrlisheim, 68021 Colmar Cedex, France1.
Department of Plant Pathology, Pennsylvania State University, University Park, PA 16802, USA2.
J Gen Virol. 2001 Aug;82(Pt 8):1995-2007. doi: 10.1099/0022-1317-82-8-1995.
Beet western yellows virus (BWYV), family Luteoviridae, is an icosahedral plant virus which is strictly transmitted by aphids in a persistent and circulative manner. Virions cross two cellular barriers in the aphid by receptor-based mechanisms involving endocytosis and exocytosis. Particles are first transported across intestinal cells into the haemolymph and then across accessory salivary gland cells for delivery to the plant via saliva. We identified the midgut part of the digestive tract as the site of intestinal passage by BWYV virions. To analyse the role in transmission of the minor capsid component, the readthrough (RT) protein, the fate of a BWYV RT-deficient non-transmissible mutant was followed by transmission electron microscopy in the vector Myzus persicae. This mutant was observed in the gut lumen but was never found inside midgut cells. However, virion aggregates were detected in the basal lamina of midgut cells when BWYV antiserum was microinjected into the haemolymph. The presence of virions in the haemolymph was confirmed by a sensitive molecular technique for detecting viral RNA. Thus, transport of the mutant virions through intestinal cells occurred but at a low frequency. Even when microinjected into the haemolymph, the RT protein mutant was never detected near or in the accessory salivary gland cells. We conclude that the RT protein is not strictly required for the transport of virus particles through midgut cells, but is necessary for the maintenance of virions in the haemolymph and their passage through accessory salivary gland cells.
甜菜西方黄化病毒(BWYV)属于黄症病毒科,是一种二十面体植物病毒,通过蚜虫以持久性循环方式严格传播。病毒粒子通过基于受体的机制穿过蚜虫体内的两个细胞屏障,该机制涉及内吞作用和外排作用。粒子首先通过肠道细胞运输到血淋巴中,然后穿过副唾液腺细胞,通过唾液传递给植物。我们确定消化道的中肠部分是BWYV病毒粒子肠道通过的部位。为了分析次要衣壳成分即通读(RT)蛋白在传播中的作用,通过透射电子显微镜在载体桃蚜中追踪了BWYV RT缺陷型非传播性突变体的命运。在肠道腔中观察到了这种突变体,但从未在中肠细胞内发现。然而,当将BWYV抗血清显微注射到血淋巴中时,在中肠细胞的基膜中检测到了病毒粒子聚集体。通过一种检测病毒RNA的灵敏分子技术证实了血淋巴中存在病毒粒子。因此,突变病毒粒子通过肠道细胞的运输发生了,但频率很低。即使显微注射到血淋巴中,在副唾液腺细胞附近或内部也从未检测到RT蛋白突变体。我们得出结论,RT蛋白对于病毒粒子通过中肠细胞的运输并非严格必需,但对于病毒粒子在血淋巴中的维持及其通过副唾液腺细胞的过程是必需的。