DETECTION OF BETA-TUBULIN IN TOMATO SEEDS - OPTIMIZATION OF EXTRACTION AND IMMUNODETECTION

SDS-PAGE profiles of SDS-denaturated total protein (MODIL) extracts of imbibing tomato seeds showed no large qualitative differences from th ose of water-soluble (HEPES) proteins. However, the protein concentrat ions of the HEPES samples were significantly lower than the MODIL ones . The presence of SDS and DTT in MODIL buffer at an alkaline pH were e ssential for high yields of beta-tubulin. The absence of DTT in HEPES extraction buffer may have allowed oxidation and subsequent loss of be ta-tubulin extracted from root tips of germinated seeds. Optimization of the beta-tubulin extraction procedure and improvements of the immun odetection system enabled us to demonstrate the beta-tubulin accumulat ion pattern in whole tomato seeds and in the different tissues. beta-T ubulin accumulation in seeds was shown to be tissue-dependent, the hig hest concentration being found in embryo root tip tissue. (C) 1998 Els evier Science Ltd. All rights reserved.