Verticillium wilt of alfalfa (Medicago sativa) is a widespread and destructive disease caused by the soil-borne fungal pathogen Verticillium alfalfae (formerly V. albo-atrum before 2011). Owing to an inadequate understanding of the pathogenicity, systemic colonisation, and host range of V. alfalfae, it has been challenging to develop an effective control measure against this disease. In the present study, seven inoculation methods, including seed inoculation, leaf spraying (LS), fungal plugs placed on leaves (FP), stem injection (SI), root dipping (RD), root injuring and dipping (RI), and watering conidia suspension into soil (WI) were used to analyse the pathogenicity and systemic colonisation of V. alfalfae on alfalfa. The typical verticillium wilt symptoms including V-shaped necrosis of leaves and leaf wilting were observed in alfalfa plants with all inoculation methods. The shortest incubation period (9 days) was observed with stem injection and the longest (40 days) with root dipping. Alfalfa plants inoculated by stem injection showed the highest disease incidence (91%), disease index (65%), and percentage of infected plants (80%), which resulted in the highest shoot biomass loss rate (34%). However, the plants inoculated by fungal plugs showed the lowest disease index (4%), percentage of infected plants (10%), and root biomass loss rate (2%). Further, the stem injection method was used to assess the host range of V. alfalfae on seven non-alfalfa plant species including erect milkvetch, sainfoin, common vetch, sunflower, potato, cotton, and bluish dogbane. The artificial inoculation of V. alfalfae by stem injection successfully infected these plants, with disease incidence ranged from 13 to 100% and disease index ranged from 10 to 69. This indicates that host range of V. alfalfae is not only alfalfa, but also other plant species.

苜蓿黄萎病（Medicago sativa）是由土壤传播的真菌病原体苜蓿黄萎病（2011年之前的原名V. albo - atrum）引起的一种广泛且具有破坏性的病害。由于对紫花苜蓿的致病性、系统定植和宿主范围的了解不足，开发针对这种疾病的有效控制措施一直具有挑战性。在本研究中，七种接种方法，包括种子接种、喷叶 (LS)、置于叶片上的真菌栓塞 (FP)、茎注射 (SI)、浸根 (RD)、伤根浸渍 (RI) 和浇水分生孢子悬浮到土壤（WI）中用于分析致病性和系统定植V. 苜蓿上的苜蓿。在所有接种方法的苜蓿植株中均观察到典型的黄萎病症状，包括叶片V形坏死和叶片萎蔫。茎注射观察到的潜伏期最短（9 天），浸根观察到的潜伏期最长（40 天）。通过茎注射接种的苜蓿植株表现出最高的发病率 (91%)、疾病指数 (65%) 和受感染植物的百分比 (80%)，从而导致最高的枝条生物量损失率 (34%)。然而，接种真菌栓塞的植物表现出最低的疾病指数 (4%)、受感染植物的百分比 (10%) 和根生物量损失率 (2%)。此外，茎注射法用于评估苜蓿的寄主范围七种非苜蓿植物物种，包括直立紫云英、红豆草、普通紫云英、向日葵、马铃薯、棉花和蓝狗根。苜蓿茎秆注射人工接种成功侵染这些植株，发病率为13%～100%，病害指数为10～69。这说明苜蓿的寄主范围不仅是苜蓿，还有其他植物。植物品种。