Structure and Expression of a Dhurrinase (-Glucosidase) from Sorghum¹

Muzaffer Cicek and Asim Esen^*

Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24060-0406

Sorghum (Sorghum bicolor L. Moench) has two isozymes of the cyanogenic -glucosidase dhurrinase: dhurrinase-1 (Dhr1) and dhurrinase-2 (Dhr2). A nearly full-length cDNA encoding dhurrinase was isolated from 4-d-old etiolated seedlings and sequenced. The cDNA has a 1695-nucleotide-long open reading frame, which codes for a 565-amino acid-long precursor and a 514-amino acid-long mature protein, respectively. Deduced amino acid sequence of the sorghum Dhr showed 70% identity with two maize (Zea mays) -glucosidase isozymes. Southern-blot data suggested that -glu-cosidase is encoded by a small multigene family in sorghum. Northern-blot data indicated that the mRNA corresponding to the cloned Dhr cDNA is present at high levels in the node and upper half of the mesocotyl in etiolated seedlings but at low levels in the rootonly in the zone of elongation and the tip region. Light-grown seedling parts had lower levels of Dhr mRNA than those of etiolated seedlings. Immunoblot analysis performed using maize-anti--glucosidase sera detected two distinct dhurrinases (57 and 62 kD) in sorghum. The distribution of Dhr activity in different plant parts supports the mRNA and immunoreactive protein data, suggesting that the cloned cDNA corresponds to the Dhr1 (57 kD) isozyme and that the dhr1 gene shows organ-specific expression.

Plant Physiol. (1998) 116: 1469-1478
Copyright Clearance Center:   0032-0889/98/116/1469/10
© 1998 American Society of Plant Physiologists

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