Institute for Sustainable Plant Protection, National Research Council of Italy (CNR), Bari, Italy.
Methods Mol Biol. 2024;2756:71-101. doi: 10.1007/978-1-0716-3638-1_2.
Once a nematode has been identified, to conduct studies for screening programs or pathogenicity tests, it is necessary a supply of large numbers of nematodes from field crops or reproduced and stored to be used in periods of the year when they are not available from fields. Therefore, nematodes must be reared in greenhouse or under in vitro conditions and stored for future needs. In this chapter, suggestions are given on how to obtain nematodes from fields and reproduce most of them on host plants in greenhouse (mainly Meloidogyne spp. and Globodera spp.) or in vitro. Reproductions in vitro include: On suitable callus of host plants (Pratylenchus spp., Ditylenchus spp.) On fungal cultures mainly of Botrytis cinerea or Alternaria spp. for Aphelenchoides spp. and other aphelenchids, including Bursaphelenchus xylophilus. On carrot disks for Pratylenchus spp. and Ditylenchus spp. Other specific media, such as garlic, potato, and sweet potato for D. destructor, and cocoyam disks for Radopholus similis. Guidelines are also given to store different nematodes for rather long times, including in vitro methods and in infected seeds, hay, and other plant parts. No information is given on how to prepare and store fixed materials.
一旦鉴定出线虫，为了进行筛选计划或致病性测试的研究，有必要从田间作物中获得大量线虫，或者进行繁殖和储存，以便在田间无法获得时在一年中的特定时期使用。因此，必须在温室或体外条件下饲养线虫，并储存以备将来使用。本章介绍了如何从田间获取线虫，并在温室（主要是根结线虫属和马铃薯金线虫属）或体外条件下在宿主植物上繁殖大多数线虫（主要是根结线虫属和马铃薯金线虫属）。体外繁殖包括：在合适的宿主植物愈伤组织上（滑刃线虫属和茎线虫属）在真菌培养物上，主要是 Botrytis cinerea 或 Alternaria spp.，用于 Aphelenchoides spp. 和其他滑刃线虫属，包括松材线虫。在胡萝卜片上用于繁殖滑刃线虫属和茎线虫属。其他特定培养基，如大蒜、土豆和甘薯用于防治毁灭茎线虫属，以及芋艿片用于防治香蕉穿孔线虫属。还提供了储存不同线虫的指南，包括体外方法以及在感染的种子、干草和其他植物部分中储存。关于如何准备和储存固定材料的信息未予提供。
