Infection Assay of Cyst Nematodes on Arabidopsis Roots
拟南芥根部的胞囊线虫感染实验
发布: 2015年09月20日第5卷第18期 DOI: 10.21769/BioProtoc.1596 浏览次数: 12601
评审: Zhaohui LiuDaniel F. CaddellAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Sep 2014
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Abstract
Plant parasitic nematodes are devastating pests on many crops. Juveniles (J2) of cyst nematodes invade the roots to induce a syncytium. This feeding site is their only source of nutrients. Male nematodes leave the roots after the fourth molt to mate with females. The females stay attached to their syncytia throughout their life and produce hundreds of eggs, which are contained in their bodies. When the females die their bodies form the cysts, which protect the eggs. Cysts can survive for many years in the soil until favorable conditions induce hatching of the juveniles.
The beet cyst nematode Heterodera schachtii is a pathogen of sugar beet (Beta vulgaris) but can also complete its life cycle on Arabidopsis roots growing on agar plates under sterile conditions. We present here protocols for a stock culture of H. schachtii and an infection assay on agar plates.
Cyst nematode (囊肿线虫)
Heterodera schachtii (大豆胞囊线虫schachtii)
Arabidopsis (拟南芥)
Materials and Reagents
- Heterodera schachtii
- Seeds of Sinapis alba cv Albatros
- Seeds of Arabidopsis thaliana
- Calcium hypochlorite
- HgCl2
- Tween 20
- 70% ethanol
- GAMBORG B5 VITAMIN MIXTURE (Duchefa Biochemie, catalog number: G0415 )
- Daichin agar (Duchefa Biochemie, catalog number: D1004 )
- Saccharose
- KNO3
- MgSO4.7H2O
- Ca(NO3)2.4H2O
- KH2PO4
- FeNaEDTA
- H3BO3
- MnCl2 or MnCl2.4H2O
- CuSO4.5H2O or CuSO4.2H2O
- ZnSO4.7H2O
- CoCl2.6H2O
- H2MoO4 or NaMoO4.2H2O
- NaCl
- ZnCl2
- GELRITETM (Duchefa Biochemie, catalog number: G1101 )
- Plastic Petri dishes 9 cm
- Plastic Petri dishes 14.5 cm
- Aluminum foil
- PVC membrane 100 µm mesh (Buddeberg GmbH, catalog number: 9068289 )
- PVC membrane 15 µm mesh (Buddeberg GmbH, catalog number: 9068280 )
- 50 ml syringes
- Pipettes
- Preparation of water agar plates (see Recipes)
- Preparation of Knop medium (see Recipes)
Equipment
- Nescofilm (or Parafilm)
- Growth chamber at 25 °C with 16 h light/8 h dark
- Laboratory glassware
- Balance
- Stereo microscope
- Inverse microscope with camera
- Clean bench
- Autoclave capable of reaching 120 °C
- Axiovert 200M inverse microscope (ZEISS) with an integrated camera (ZEISS, model: AxioCam MRc5 )
- Funnel for hatching (Figure 1)
Software
- Contour tool of the AxioVision software (ZEISS)
- SPSS 12.0 (SPSS Inc.)
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Bohlmann, H. and Wieczorek, K. (2015). Infection Assay of Cyst Nematodes on Arabidopsis Roots. Bio-protocol 5(18): e1596. DOI: 10.21769/BioProtoc.1596.
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分类
微生物学 > 微生物-宿主相互作用 > 体外实验模型
微生物学 > 微生物-宿主相互作用 > 线虫
植物科学 > 植物免疫 > 病害生物测定
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