CBN-V Video Archives - S5-04
Analysis of Proteins Associated with Storage Root Formation in Cassava using 2-D Gels

Cabral, G.B. and L.J.C.B. Carvalho

EMBRAPA-Genetic Resources and Biotechnology, CP 02372, CEP 70770-900 Sain Parque Rural, Brazilia, Brazil carvalho@cenargen.embrapa.br

        Two-dimensional gel electrophoresis analysis was performed on adventitious and storage roots of cassava (Manihot esculenta Crantz). Non-swollen adventitious roots and swollen storage root were obtained from the accession WU104 grown in the greenhouse at the Department of Biology, Washington University, St. Louis, MO-USA. Saline buffer-soluble proteins were extracted, separated in a high-resolution 2-D electrophoresis system and visualized using silver staining gel procedure. Digital images were generate for further analysis. Quantitative and qualitative protein spots analysis was performed with a computer assisted image software system. Results revealed large variation in the complexity of the gel protein profile between the two root systems. About 90% of the protein spots appeared in the pI range value of 4.0 to 6.5 and between 14 to 80 Kda of molecular mass. Detailed computer assisted analysis of this gel allowed us to establish 5 distinct classes of protein based on spot quantification that could be associated with swelling and non-swelling roots. Variation in the complexity of protein pattern was related with different type of root. Adventitious roots showed a more simple profile related to primary growth, while the storage roots displayed to be a more complex profile related to secondary growth and starch accumulation.

This research financed by the National Biotechnology Program (EMBRAPA), and The Rockefeller Foundation (USA)

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