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Plant Physiol, March 2001, Vol. 125, pp. 1396-1405

Identification of Mutator Insertional Mutants of Starch-Branching Enzyme 2a in Corn1

Susan L. Blauth,2 Yuan Yao, Jeffery D. Klucinec, Jack C. Shannon, Donald B. Thompson, and Mark J. Guiltinan*

The Biotechnology Institute (S.L.B., M.J.G.) and Departments of Food Science (Y.Y., J.D.K., D.B.T.) and Horticulture (J.C.S., M.J.G.), Pennsylvania State University, University Park, Pennsylvania 16802

Starch-branching enzymes (SBE) break the alpha -1,4 linkage of starch, re-attaching the chain to a glucan chain by an alpha -1,6 bond, altering starch structure. SBEs also facilitate starch accumulation by increasing the number of non-reducing ends on the growing chain. In maize (Zea mays), three isoforms of SBE have been identified. To examine the function of the SBEIIa isoform, a reverse genetics polymerase chain reaction-based screen was used to identify a mutant line segregating for a Mutator transposon within Sbe2a. To locate the insertion within the second exon of Sbe2a, the genomic sequence of Sbe2a containing the promoter and 5' end was isolated and sequenced. Plants homozygous for sbe2a::Mu have undetectable levels of Sbe2a transcripts and SBEIIa in their leaves. Characterization of leaf starch from sbe2a::Mu mutants shows reduced branching similar to yet more extreme than that seen in kernels lacking SBEIIb activity. Characterization of endosperm starch from sbe2a::Mu mutants shows branching that is indistinguishable from wild-type controls. These mutant plants have a visible phenotype resembling accelerated senescence, which was correlated with the Mutator insertion within Sbe2a. This correlation suggests a specific role for SBEIIa in leaves, which may be necessary for normal plant development.

1 This work was supported by the U.S. Department of Energy, Bioscience Program (grant no. DE-FG02-96ER20234 to M.J.G., J.C.S., and D.B.T.) and by the Pennsylvania State Agricultural Experiment Station (grant no. CRIS 3303).

2 Present address: Department of Biology, University of Redlands, Redlands, CA 92373.

* Corresponding author; e-mail mjg9{at}psu.edu; fax 814-863-1357.

© 2001 American Society of Plant Physiologists


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