一种番茄根结线虫人工培养与保存优化方法.doc

一种番茄根结线虫人工培养与保存优化方法 　　摘 要：根结线虫病在世界范围内对粮食蔬菜水果产量影响严重，其人工培养具有依赖寄主植物的局限性。从山东省泰安市房村镇的日光温室中采集被根结线虫侵染严重的番茄根系，用机械捣碎-过筛喷淋法从根系样品中分离根结线虫和卵，以牡丹根腐病镰刀菌（Fusarium solani）制作二重培养基，分别接种根结线虫悬液与卵悬液培养。结果显示，接种线虫悬液与卵悬液的平板分别在第三周与第四周线虫密度达到最高峰，培养基中虫口密度分别为2 300头/ml与2 000头/ml。置培养平板于15℃条件下存放，5个月后培养基内仍有可供继代繁殖的活虫体。该优化后的方法脱离了寄主植物束缚，简化了培养过程，具有高效繁殖和长期保存根结线虫的优点。 关键词：根结线虫；人工培养；牡丹根腐病镰刀菌；优化方法 中图分类号：S436.412.2+9 文献标识号：A 文章编号：1001-4942（2012）11-0117-04 An Optimized Artificial Culture and Preservation Method for Tomato Root-knot Nematodes Liu Li， Yan ShiCui， Yao LiangTong， Ding YanQin， Du BingHai* （College of Life Science， Shandong Agricultural University /Shandong Key Laboratory of Agricultural Microbiology， Taian 271018， China） Abstract Root-knot nematodes seriously decreased the production of crops， fruits and vegetables worldwide， and its artificial culture relied heavily on host plants. In this study， the tomato roots infected by root-knot nematodes were adopted from greenhouse of Fangcun Town， Taian City， Shandong Province， and the root-knot nematodes and eggs were separated by the method of mechanical beating-sifting spray and inoculated to double medium of Fusarium solani. The density of root-knot nematodes was about 2 300 per mini liter medium when inoculated with root-knot nematodes for three weeks and about 2 000 when inoculated with eggs for four weeks. Live root-knot nematodes could still be obtained after 5 months， if the culture plate was preserved at 15℃. This optimized method was independent of host plants， processed more simply， could obtain large quantity of root-knot nematodes in short time and preserve for long time. Key words Root-knot nematode； Artificial culture； Fusarium solani； Optimized method 根结线虫（Meloidogyne spp）是一类广泛分布在世界各地的植物根系定居性内寄生线虫[1]。根结线虫属寄主范围广泛，可侵染的植物超过3 000种，分属114科，如蔬菜、粮食作物、经济作物和果树、观赏植物以及杂草都可被侵染[2，3]。根结线虫一旦发生，很难彻底清除，连作期愈长发病愈重。在温带、亚热带、热带地区的植物受害比其他地区更为严重，病害可造成作物减产10%～20%，严重时可达75%以上，还能传播一些真菌和细菌性病害[4，5]。 目前关于根结线虫的分类、生物防治、基因组研究已比较深入，而其培养和保存的研究