title: Cryopreservation of Radopholus similis, a tropical plant-parasitic nematode.
abstract: For obligate plant-parasitic nematodes, cryopreservation has advantages over the usual preservation methods on whole plants or axenic culture systems, because the latter two are labourious and time and space consuming. In addition, cross contamination among different isolates can occur easily. Moreover, specific genetic studies require maintenance of the original population. The nematode under investigation, Radopholus similis, is a plant-parasitic nematode from the humid tropics. Therefore, any treatment at low temperatures is likely to add extra stress to the nematode, making the development of a cryopreservation protocol extremely difficult. In this paper, we describe experiments to achieve a successful cryopreservation protocol for the tropical nematode R. similis using vitrification solution-based methods based on a well defined mixture of cryoprotectants in combination with ultra-rapid cooling and thawing rates. A two-step treatment was used consisting of an incubation in glycerol followed by the application of a vitrifying mixture of methanol, glycerol and glucose. After cryopreservation, the pathogenicity of the nematodes was not altered, since they could infect and reproduce on carrot discs after recovery in the Ringer solution. The cryopreservation method described can be used for routine cryopreservation of R. similis lines from different origins.
authors: Laboratory of Tropical Crop Improvement, Department of Biosystems, Faculty of Bioscience Engineering, KU Leuven, Kasteelpark Arenberg 13, 3001, Leuven, Belgium.
authors: Annemie
authors: A
authors: Elsen
authors: Salvador Ferrandis
authors: SF
authors: Vallterra
authors: Tom
authors: T
authors: Van Wauwe
authors: Trinh Thi Thu
authors: TT
authors: Thuy
authors: Rony
authors: R
authors: Swennen
authors: Dirk
authors: D
authors: De Waele
authors: Bart
authors: B
authors: Panis
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D005615:Freezing; D005947:Glucose; D005990:Glycerol; D000432:Methanol; D010944:Plants / Q000469:parasitology; D012997:Solvents; D013997:Time Factors; D014329:Tropical Climate; D017187:Tylenchida / Q000254:growth & development / Q000502:physiology
chemical_list: D003451:Cryoprotective Agents; D012997:Solvents; D005947:Glucose; D005990:Glycerol; D000432:Methanol