Allelic variation and differential expression of methionine-rich delta-zeins in maize inbred lines B73 and W23a1.

Abstract

The sulfur-amino-acid-rich delta-zeins of maize ( Zea mays L.) are represented by 18-kDa and 10-kDa proteins. We have cloned a novel 11-kDa methionine-rich delta-zein from developing endosperm of the inbred line W23a1. The nucleotide sequence of this new delta-zein is identical to the published 10-kDa delta-zein, except for an insertion of 18 nucleotides between +316 and +333 bp from the translation start site. Antibodies raised against the recombinant 18-kDa delta-zein recognized both the 18-kDa and 10-kDa delta-zein from total seed protein extracts of different maize inbred lines. Western blot analysis revealed differences in the levels of the delta-zeins in different inbred lines and some of the inbred lines lacked either the 10-kDa or the 18-kDa delta-zeins. Northern blot analysis revealed temporal differences in the RNA transcript levels of the 11-kDa and 18-kDa delta-zeins between B73 and W23a1. Such differences were not evident on Western blot analysis where similar protein accumulation profiles were seen for both lines. Immunostaining of paraffin sections of developing maize endosperm with the 18-kDa delta-zein antibodies revealed specific labeling of protein bodies found in the first few starchy layers from the aleurone layer. Electron-microscopic observation of thin-sections of B73 and W23a1 endosperm cells confirmed the presence of recently discovered novel, vacuole-like structures in these inbred lines. Immunogold labeling studies revealed that the delta-zeins were localized in the endoplasmic-reticulum-derived protein bodies and showed no preferential gold particle labeling over either the light or electron-dense material found in these protein bodies.