In vitro Nematode Infection on Potato Plant
体外线虫侵染马铃薯实验
发布: 2014年01月05日第4卷第1期 DOI: 10.21769/BioProtoc.1016 浏览次数: 14448
评审: Tie Liu
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参见作者原研究论文
The authors used this protocol in:
Apr 2013
Abstract
Potato cyst nematodes (PCNs; Globodera rostochiensis and G. pallida) are devastating pests that infect potato root. We describe an in vitro assay for PCN infection on potato plantlet in tissue culture. This method is useful for studying nematode parasitism on potato and for investigating responses of potato clones/lines to PCN infection.
Keywords: Potato cyst nematode (线虫)
Globodera rostochiensis (马铃薯金线虫)
Nematode infection (线虫感染)
Potato plantlet (马铃薯试管苗)
Materials and Reagents
- Potato plant (Solanum tuberosum cv. Désirée)
- Potato cyst nematode (G. rostochiensis) cysts
- Sodium azide (RICCA Chemical, catalog number: 7144-16 )
- Mercuric chloride (RICCA Chemical, catalog number: 4650-16 )
- Sterile distilled water
- Gentamycin (Fisher Scientific, catalog number: 61398-0010 )
- Nystatin (Sigma-Aldrich, catalog number: N-3503 )
- Agarose (Fisher Scientific, catalog number: BP160-100 )
- Timentin (PhytoTechnology Laboratories®, catalog number: T869 )
- Micropore tape (Fisher Scientific, catalog number: 19-027-761 )
- Sterilization solution (0.004% mercuric chloride/0.004% sodium azide)
- Hoagland's solution (Sigma-Aldrich, catalog number: H2395 )
- Amberlite XAD-4 resin (Fisher Scientific, catalog number: AC20223 )
- MS salt (Caisson Laboratories, catalog number: MSP01-50LT )
- Inositol (Fisher Scientific, catalog number: AC122261000 )
- Thiamine HCl (Sigma-Aldrich, catalog number: T1270 )
- Gelrite (Fisher Scientific, catalog number: CAS 71010-52-1 )
- Potato root diffusate (PRD) (see Recipes)
- Propagation medium (see Recipes)
- 0.1% agarose (see Recipes)
Equipment
- Sieves No. 60 (250 μm), No. 200 (75 μm) and No. 500 (25 μm) (Endecotts, catalog number: 683927 , 681837 , 691131 )
- Sterile nematode egg hatching chamber (stainless steel metal pan cover and collection pan may be purchased from Vollrath Products, size six inches; the metal screened pan cover with wire-screen in the central area as shown in Figure 3 was custom made)
- Sterile 6-well plates (Greiner Bio-One GmbH, catalog number: 657185 )
- Stirplate
- Sterile forceps and scalpel
- Growth incubator
- 30-μm mesh (Small Parts, catalog number: 7050-1220-000-12 )
- Tissue culture biosafety cabinet
- Centrifuge
- Incubator shaker (New Brunswick Scientific, model: C24KC )
- S. tuberosum growth chambers (we use several types of growth chambers such as I-66LLVL from Percival)
- Rubber stopper (size no. 1) (Fisher Scientific, catalog number: 14-130C )
- Beaker
- Aluminum foil
- Dissecting microscope
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Chronis, D., Chen, S., Lang, P., Tran, T., Thurston, D. and Wang, X. (2014). In vitro Nematode Infection on Potato Plant. Bio-protocol 4(1): e1016. DOI: 10.21769/BioProtoc.1016.
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分类
植物科学 > 植物免疫 > 病害生物测定
细胞生物学 > 细胞分离和培养 > 细胞生长
植物科学 > 植物生理学 > 植物生长
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