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Lack of a conclusive evidence of ingestion of plastid components by plant-parasitic nematodes cautions the use of plastid transformation technology for nematode resistance. Nematode-resistant effector proteins generally require ingestion by the nematode to be effective. Transgene-encoded proteins produced in plastids are not known to be exported into the cytoplasm. Disintegration of plant cell organelles after nematode feeding suggests that nematodes possibly ingest plastid components. Proof of ingestion will validate the development of plastid transformation for nematode resistance. Small subunit ribulose-1,5-bisphosphate carboxylase/oxygenase (SSRubisco) protein is prevalent in chloroplasts and thus chosen as a study molecule. The migratory endoparasitic nematodes  Pratylenchus penetrans  and  Radopholus similis  were cultured on green carrot callus containing chloroplasts. Total nematode proteins were extracted and subjected to western blot analysis using cross-reactive polyclonal antibody raised against spinach SSRubisco. Positive detection of SSRubisco occurred in protein extracts from nematodes fed on green carrot callus, but not in extracts from nematodes cultured on non-green alfalfa root callus as negative control. These results confirm the ingestion of plastid components of MW 14 kDA by migratory endoparasitic nematodes.

(287) Chloroplast Small Subunit Rubisco Protein Is Ingested by Migratory Endoparasitic Nematodes