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ARS Home » Pacific West Area » Corvallis, Oregon » Horticultural Crops Research » Research » Research Project #435571

Research Project: Eradication Strategies for Globodera pallida, the Pale Cyst Nematode (PCN)

Location: Horticultural Crops Research

Project Number: 2072-22000-043-33-I
Project Type: Interagency Reimbursable Agreement

Start Date: Jul 1, 2018
End Date: Dec 30, 2019

Objective:
1. Optimization of mustard seed meal extracts to maximize PCN eradication. 2. Metagenomics of microbial community of Globodera

Approach:
The active ingredient precursor glucosinolate from B. juncea seed meal will be extracted with hot water using a stirred reactor, one of the components integrated into our recently constructed pilot plant. S. alba seed meal will be extracted with same reactor, but at lower temperatures that will promote glucosinolate hydrolysis. We will determine by what percentage re-extraction of the spent meal will improve active ingredient yield. Solids will be separated using a large scale decanter and the aqueous extract containing 20% solids will be dried using a spray dryer or freeze dryer. The most efficient and cost effective drying method will be identified. Emphasis will be placed on developing large scale extraction procedures that will generate extract volumes sufficient for extensive field testing, and on developing processing protocols that can be further scaled up in a commercial operation. The concentrated extract will be formulated with a catalyst required for the release of bioactive compounds at the time of application. DNA will be isolated from Globodera eggs collected from infested fields. Cysts will be suspended in water and disrupted by using a pestle in a microfuge tube. The eggs will be collected and will be suspended in a DNA extraction buffer containing Proteinase K enzyme and lysed by using a TissueLyser. DNA will be purified using a binding matrix (using a standard kit protocol). Universal adapter primers for fungi and bacteria will be used for preparation of sequencing libraries. Specific primers for nematode-trapping fungi (Orbiliales) will also be used to identify the diversity and presence of this group of fungi within eggs.