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Biology
マツノザイセンチュウ Bursaphelenchus xylophilus によるIn vivoおよびIn vitroマツの感染と誘...
マツノザイセンチュウ Bursaphelenchus xylophilus  によるIn vivoおよびIn vitroマツの感染と誘...
JoVE Journal
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JoVE Journal Biology
Infection of In Vivo and In Vitro Pines with the Pinewood Nematode Bursaphelenchus xylophilus and Isolation of Induced Volatiles

マツノザイセンチュウ Bursaphelenchus xylophilus によるIn vivoおよびIn vitroマツの感染と誘導揮発性物質の単離

Full Text
1,199 Views
08:42 min
September 27, 2024

DOI: 10.3791/67149-v

Jorge M. S. Faria1,2, A. Cristina Figueiredo3,4, Dora M. Teixeira5, Maria L. Inácio1,2

1INIAV, I.P., National Institute for Agrarian and Veterinary Research, I.P., 2GREEN-IT Bioresources for Sustainability, Instituto de Tecnologia Química e Biológica,Universidade Nova de Lisboa (ITQB NOVA), 3Centre for Environmental and Marine Studies (CESAM Lisboa), Biotecnologia Vegetal (BV), DBV,Faculdade de Ciências da Universidade de Lisboa, 4Centre for Ecology, Evolution and Environmental Changes (CE3C), Biotecnologia Vegetal (BV), DBV,Faculdade de Ciências da Universidade de Lisboa, 5Hercules Laboratory & Chemistry and Biochemistry Department of Science and Technology School,University of Évora

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Overview

This study presents a detailed protocol for investigating the interaction between the pinewood nematode and Pinus pinaster, both in vitro and in vivo. It emphasizes the analysis of volatile organic compounds emitted by infected pine trees, which sheds light on the host's response to these phytoparasites.

Key Study Components

Research Area

  • Pinewood nematode infection
  • Volatile organic compound analysis
  • Plant-pathogen interactions

Background

  • The genetic diversity of pine plantlets impacts volatile profiles.
  • In vitro co-culture systems provide a controlled environment to reduce variability.
  • Understanding host responses is crucial for managing pine wilt disease.

Methods Used

  • In vitro co-culture systems of pine shoots and nematodes
  • GC and GC-MS for volatile profiling
  • Greenhouse bioassays for symptom quantification

Main Results

  • The establishment of a 100-fold increase in nematode population within eight days.
  • Signs of pine wilt disease observed 20 days post-inoculation.
  • Successful isolation of volatiles from the infected tissues.

Conclusions

  • This protocol enhances our understanding of plant responses to nematode infection.
  • The findings contribute to the broader field of plant pathology and nematology.

Frequently Asked Questions

What is the significance of studying the pinewood nematode?
Understanding its infection mechanisms can help in managing pine wilt disease.
How does genetic diversity affect volatile profiles?
Different genetic backgrounds can lead to varying responses in volatile emissions during nematode infections.
What technologies are utilized in this research?
Gas Chromatography (GC) and Mass Spectrometry (GC-MS) are essential for analyzing volatiles.
Why is the in vitro co-culture system beneficial?
It minimizes variability caused by genetic differences among plant clones, allowing clearer results.
How long does it take to observe symptoms of infection?
Symptoms of pine wilt disease can be observed approximately 20 days after nematode inoculation.
What methods are used to quantify nematode populations?
Nematodes are counted using a microscope on a concave slide.
Can the volatile compounds indicate plant health?
Yes, changes in volatile profiles can signal stress or health status related to pest infections.

このプロトコルは、Pinus pinasterのin vivoおよびin vitroのマツノイセンチュウ感染と、ガスクロマトグラフィー(GC)および質量分析(GC-MS)に結合されたGCによるそれらの揮発性分析について説明しています。

私たちは、マツノコとマツノザイセンチュウのIn Vitro共培養システムを確立し、これらの植物寄生虫に対する宿主の揮発性応答の変化を市場に出し、研究しています。温室バイオハウスは、環境変動を制御することができます。しかし、マツの苗の遺伝的多様性は、結果として生じる揮発性プロファイルに依然として影響を与えています。

マツノザイセンチュウに感染したインビトロマツの新芽は、容易に入手できる植物クローンのサンプリングを可能にし、マツの遺伝的多様性による変動の影響を軽減します。植物の揮発性物質は、蒸留抽出の水蒸留によって単離されますが、ヘッドスペース技術により、植物から放出される揮発性物質のプロファイリングが可能になります。レジン充填カラムは、マツ線虫に感染したIn Vitro粉砕マツから放出される揮発性物質をプロファイリングするための非侵襲的手法として使用され、宿主と植物寄生虫との間の化学的相互作用を直接分析します。

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