拟南芥根部的胞囊线虫感染实验

Holger Bohlmann; Krzysztof Wieczorek

doi:10.21769/BioProtoc.1596

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
CN
- EN - English
- CN - 中文

Peer-reviewed

Infection Assay of Cyst Nematodes on Arabidopsis Roots

拟南芥根部的胞囊线虫感染实验

HB Holger Bohlmann email

Krzysztof Wieczorek email

发布: 2015年09月20日第5卷第18期 DOI: 10.21769/BioProtoc.1596 浏览次数: 11646

评审: Zhaohui LiuDaniel F. CaddellAnonymous reviewer(s)

下载 PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Molecular Plant Microbe Interactions, featuring study using the protocol.

Sep 2014

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

用于研究寄主皮肤反应的巴西日圆线虫幼虫的制备

Tiffany Bouchery [...] Nicola L. Harris

2020年12月20日 2168 阅读

秀丽隐杆线虫细菌负荷的定量研究

Alyssa C. Walker [...] Daniel M. Czyz

2022年01月20日 3126 阅读

对早、中、晚草莓进行灰葡萄孢体内接种试验

Piao Yang [...] Ye Xia

2023年10月20日 1423 阅读

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.

Keywords: Nematode infection assay (线虫感染的检测)

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
GELRITE^TM (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

English

中文翻译

文章信息

版权信息

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.