Endosperm protein accumulation in wild and cultivated barley and their cross grown in spike culture

Abstract

The high protein wild relatives of cultivated cereals have proven difficult to utilize in plant breeding by direct selection for high grain protein percentage, and hence alternative selection criteria are needed. In this study, a spike culture method was used to measure differences in protein accumulation between wild and cultivated barley, and their cross, at different levels of nitrogen supply. Three genotypes, barley cultivar Hordeum vulgare L. cv. Clipper, a wild barley accession H. spontaneum Koch line 363, and a high protein F5 line (38.4) derived from their cross, were grown from 8 to 27 days after flowering in in vitro spike culture. Nitrogen supply in the culture medium was either 0.4 g/l or 2.0 g/l of N supplied as NH4NO3. Spikes were harvested at approximately 3 day intervals during grain development, and salt soluble and hordein protein fractions were measured. Lines 363 and 38.4 differed from ‘Clipper’ in having extremely high initial rates of protein accumulation, even at 0.4 g/l N. In high nitrogen conditions all three genotypes reached similar salt soluble plus hordein protein levels. Hordein-1 and hordein-2 fractions were measured separately; the percent of hordein-1 was higher in lines 363 and 38.4 than in ‘Clipper’ at 0.4 g/l N. For all parameters measured, pot-grown spikes of matching age were harvested and were shown to be similar to the 0.4 g/l N treatment. The possible utilization of spike culture for identification of critical protein accumulation parameters is discussed, in relation to their possible utilization in breeding.