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.

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Previous studies indicated that the deficiency of starch-branching enzyme (SBE) Ia in the single mutant sbe1a::Mu (sbe1a) has no impact on endosperm starch structure, whereas the deficiency of SBEIIb in the ae mutant is well known to reduce the branching of starch. We hypothesized that in maize (Zea mays) endosperm, the function of SBEIIb is predominant to that of SBEIa, and SBEIa would have an observable effect only on amylopectin structure in the absence of SBEIIb. To test this hypothesis, the mutant sbe1a was introgressed into lines containing either wx (lacking the granule-bound starch synthase GBSSI) or ae wx (lacking both SBEIIb and GBSSI) in the W64A background. Both western blotting and zymogram analysis confirmed the SBEIa deficiency in sbe1a wx and sbe1a ae wx, and the SBEIIb deficiency in ae wx and sbe1a ae wx. Using zymogram analysis, no pleiotropic effects of sbe1a genes on SBEIIa, starch synthase, or starch-debranching enzyme isoforms were observed. Highperformance size exclusion chromatography analysis shows that the chain-length profiles of amylopectin as well as b-limit dextrin were indistinguishable between wx and sbe1a wx, whereas significant differences for both were observed between ae wx and sbe1a ae wx, suggesting an effect of SBEIa on amylopectin biosynthesis that is observable only in the absence of SBEIIb. The amylopectin branch density and the average number of branches per cluster were both higher in endosperm starch from sbe1a ae wx than from ae wx. These results indicate possible functional interactions between SBE isoforms that may involve enzymatic inhibition. Both the cluster repeat distance and the distance between branch points on the short intracluster chains were similar for all genotypes however, suggesting a similar pattern of individual SBE isoforms in cluster initiation and the determination of branch point location.Starch biosynthesis involves the activities of four enzymes: ADP-Glc pyrophosphorylase, starch synthase (SS), starch-branching enzyme (SBE), and starch-debranching enzyme (DBE;Smith et al., 1997). The coordinated functions of these enzymes result in two forms of starch molecules: amylose, an essentially linear glucan chain consisting of a-1,4 linkages, and amylopectin, a highly branched glucan chain with multiple branch points formed by a-1,6 linkages.SBE catalyzes the formation of branch points within glucan chains by cleaving an a-1,4 linkage and reattaching the chain to a glucan chain via a-1,6 bond. Three isoforms of SBE (SBEIa, SBEIIa, and SBEIIb) have been identified in maize (Zea mays; Boyer and Preiss, 1978;Dang and Boyer, 1988), and their expression patterns differ considerably (Gao et al., 1996). While the in vitro properties of SBEIa differ from those of SBEIIa and SBEIIb (Guan and Preiss, 1993;Takeda et al., 1993), the sbe1a or sbe2a single mutants obtained using Mu-mediated transposon mutagenesis indicated that the chain-length (CL) profiles of starch produced in endosperm in vivo were not affected by the deficiency of either SBEIa or SBEIIa (Blau...

of alkenones in the spring transitional season, concurrent with the period of lake ice melt and isothermal mixing. Together, these data provide a framework for evaluating lacustrine alkenone distributions and utilizing alkenone unsaturation as a spring lake temperature proxy in freshwater lakes. 70

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Phytoglycogen octenyl succinate (PG-OS) and epsilon-polylysine (EPL) were used to form oil-in-water emulsions with enhanced lipid oxidative stability. PG-OS is an amphiphilic carbohydrate nanoparticle prepared using octenyl succinate (OS) substitution of phytoglycogen (PG). PG-OS had a dispersed molecular density nearly 20 times that of waxy corn starch octenyl succinate (WCS-OS). Fish oil-in-water emulsions were prepared using PG-OS, WCS-OS, and Tween 20, stored at 55 degrees C for 6 days, and monitored for the accumulation of hydroperoxide and thiobarbituric acid reactive substances (TBARS). The result indicated that PG-OS may lead to high lipid oxidative stability, and that the addition of EPL may further improve the oxidative stability of emulsions. To address the interaction between PG-OS and EPL, zeta-potential was determined for various systems. The results indicated a possible formation of an interfacial complex layer comprising both PG-OS and EPL. This complex layer may provide both physical and electrostatic barriers against pro-oxidative compounds.

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