Abstract：
Cucurbit leaf crumple virus (CuLCrV) is among the prominent viruses infecting cucurbits in the USA. Attainable procedures of virus inoculation to crops are prerequisite for screening of resistance against the virus. Because mechanical (non-vector-mediated) infection by cucurbit leaf crumple virus (CuLCrV) is inefficient in economically important crops, screening for CuLCrV resistance is currently laborious and time-consuming using transmission by viruliferous whiteflies. We constructed an infectious partial tandem repeat construct of an isolate of CuLCrV from Georgia, USA, in the plant expression binary vector pCambia2300 and transformed it into Agrobacterium tumifaciens strain EHA105. Agroinfiltration of this construct into the abaxial surface of the leaves of common bean (Phaseolus vulgaris L.) produced a systemic infection characteristic of CuLCrV, although this approach was not successful for yellow squash. However, we report a very efficient and reproducible inoculation procedure established in squash when the leaves were injured with a microneedle and rubbed it with cell suspension harbouring the infectious viral construct.
摘要：
葫芦叶皱病毒（CuLCrV）是美国感染葫芦的主要病毒之一。对作物进行病毒接种的可行程序是筛选病毒抗性的先决条件。由于葫芦叶皱病毒（CuLCrV）的机械（非媒介介导的）感染在经济上重要的作物中效率低下，筛选CuLCrV抗性目前是费力且费时的，使用含病毒粉虱的传播。我们构建了来自佐治亚州的CuLCrV分离物的感染性部分串联重复结构，美国,在植物表达二元载体pCambia2300中，并将其转化到肿瘤农杆菌菌株EHA105中。该结构向普通豆（PhaseolusvulgarisL.）叶子的背面的渗透产生了CuLCrV的全身性感染特征，尽管这种方法对于壁球并不成功。然而,我们报告了一种非常有效且可重复的接种程序，该程序在南瓜中建立，当时用微针伤害叶片并用带有感染性病毒构建体的细胞悬浮液擦拭。
