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NOTES

Trypsin inhibitory activity measurement

Simplifications of the standard procedure used for pea seed

^a Institut National de la Recherche Agronomique, Unité de Recherche en Génétique et Amélioration des Plantes, 17 rue Sully, BV1540 F21034 Dijon Cedex, France
^b Institut National de la Recherche Agronomique, Laboratoire de Biochimie et Technologie des Protéines, Rue de la Géraudière, BP 71627 F44316 Nantes Cedex 03, France

page{at}epoisses.inra.fr

A major breeding criterion for pea (Pisum sativum L.) is a low seed trypsin inhibitor activity (TIA), but the high cost of measurement limits the screening of new experimental lines. Our objectives were to devise two simplified procedures for screening for TIA using micro titration plates. In the first procedure, the inhibition induced by one single extract is measured. It allows for a rapid ranking of genotypes, but not calculation of seed TIA. Conversely, the second procedure is a titration procedure, where results are expressed in conventional trypsin inhibitor units (TIU). The time required for the measurements is reduced compared with that required with the standard procedures. The different methods were performed on cultivars ranging from low to high seed TIA. The results using the screening procedure showed very low standard deviations and allowed the ranking of the cultivars into three classes according to their seed TIA. The ranking was consistent with results obtained using the standard procedures. Results of the titration procedure were not significantly different from results obtained with the standard procedures on pea lines ranging from 2 to 10 TIU mg^-1 dry weight.

Abbreviations: AFNOR, French Association for Normalization • AOCS, American Oil Chemists' Society • BAPA, benzoyl-DL-arginine-p-nitroaniline • I%, percentage of inhibition • TI, trypsin inhibitor • TIA, trypsin inhibitor activity • TIU, trypsin inhibitor unit