A Spray Inoculation Technique to Establish Fungal Infection in Rice Plant Leaves

Abstract

Source: Zhang, M., et al. The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea. J. Vis. Exp. (2018).

This video demonstrates a technique to establish the pathogenic fungus Magnaporthe grisea infection in rice plant leaves. The fungal conidia suspension is sprayed on sections of rice leaf, and the disease progression is assessed by scoring the formation of necrotic lesions.

Protocol

1. Spray Inoculation with a Suspension of M. grisea Conidia

1. Fungal culture for M.grisea
   1. Prepare the oatmeal tomato agar (OTA) culture medium for fungal strains.
   2. Weigh 30 - 50 g of oatmeal, add this to 800 mL of distilled/deionized water (ddH₂O), and boil the mixture for 30 min in the electric pot.
   3. Filter the boiled oatmeal juice into the beaker through a piece of gauze.
   4. Add 150 mL of tomato juice and 20 g of agar to the filtrate in the beaker and add ddH₂O up to 1,000 mL.
2. Preparation of experimental material....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Agar	AOBOX Biotechnology (China)	01-023
Filter paper	GE Healthcare brand(Sweden)	10311387
50-mL tube	CORNING(Amercia)	430290
Centrifuge	Eppendorf(Amercia)	5804R
Tween-20	Coolaber (China)	CT11551-100ml
Culture dish	Thermofisher(Amercia)	150326
0.5-5 mL pipette	Eppendorf	4920000105
100-1000uL pipette	Eppendorf	4920000083
Vacuum pump	Leybold	D25B
Incubator	MEMMERT	PYX313
Inoculation ring	Greiner Bio One	731175