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Biology

アグロ浸潤およびPVXアグロ感染ジャガイモと doi: 10.3791/50971 Published: January 3, 2014

Abstract

アグロ浸潤およびPVXのアグロ感染植物における候補遺伝子の機能解析のための効率的な2一過性発現アッセイである。アグロインフィルトレーションのために最も一般的に使用される薬剤は、 アグロバクテリウム·ツメファシエンス、多くの双子葉植物種の病原体である。これは、アグロ浸潤は、多くの植物種に適用することができることを意味する。ポテト( ジャガイモ )にこれらの方法を適用する際に、ここで、我々はプロトコルと期待される結果を発表、それに関連する野生塊茎有利子ナス種ナスセクションPetota)とモデル植物ベンタミアーナ 。このような抵抗(R)または非病原性( のAvr)遺伝子のような単一遺伝子の機能解析に加えて、アグロ浸潤アッセイは、単純にRのAvr導入遺伝子を送達することによって、特定の宿主病原体相互作用に関連したR-AVR相互作用をrecapitulatingするのに非常に適している同じセル。しかし、いくつかの植物GEnotypesは、我々はいくつかのジャガイモの遺伝子型については、例えば観察されるように、 アグロバクテリウムへの非特異的な防御反応を発生させることができます。アグロ浸潤と比較して、PVXのアグロ感染とAVR活性の検出は、機能的スクリーニングにおいて、より高スループット、より敏感であり、 アグロバクテリウムへの非特異的な防御反応の影響を受けにくい。しかし、PVXに非特異的な防御が発生する可能性がありますし、ウイルス誘導極端な抵抗による応答を欠場する危険性があります。このような制限があるにもかかわらず、我々の経験では、アグロインフィルトレーションおよびPVXのアグロ感染が互いの結果を確認するために同時に使用することができ、両方の適切かつ補完的なアッセイである。

Materials

Name Company Catalog Number Comments
Beef extract Sigma-Aldrich B4888
Bacteriological peptone Oxoid LP0037
Yeast extract Oxoid LP0021
MgSO4 Sigma-Aldrich 208094
MS salts (without vitamins) Duchefa Biochemie M0221
MES Duchefa Biochemie M1503
LB broth powder Sigma-Aldrich L3022
Acetosyringone Sigma-Aldrich D134406
Syringe (1 ml) BD Plastipak 300013
Incubator Infors HT Multitron II
Centrifuge Heraeus Multifuge 3S-R
Spectrophotometer Eppendorf Biophotometer 6131

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アグロ浸潤およびPVXアグロ感染ジャガイモと<em&gt;ベンサミアナタバコ</em
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Cite this Article

Du, J., Rietman, H., Vleeshouwers, V. G. A. A. Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana. J. Vis. Exp. (83), e50971, doi:10.3791/50971 (2014).More

Du, J., Rietman, H., Vleeshouwers, V. G. A. A. Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana. J. Vis. Exp. (83), e50971, doi:10.3791/50971 (2014).

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