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A recently developed wheat variety, known as 5-5 wheat, which has inactive GBSSI-B1, GBSSI-D1, SSIIa-B1, and SSIIa-D1 isozymes, accumulates a novel type of starch, affecting bread texture and leading to reduction in bread staling. These properties are potentially useful for commercial bakery products; thus, the 5-5 genotype represents a new resource for wheat breeding. In this study, the 5-5 alleles were backcrossed into the hard wheat variety "Minaminokaori" and the soft wheat variety "Shirogane-Komugi", which are both leading Japanese wheat varieties. In comparison to their parental varieties, the two 5-5 near-isogenic lines (NILs) showed a decrease in amylose levels, an increase in the proportion of short chains of amylopectin, a lower gelatinization temperature and enthalpy change, a higher peak viscosity and breakdown viscosity as measured by a Rapid Visco Analyzer, a reduced retrogradation rate, and an increase in grain hardness. Importantly, the 5-5 NILs also showed lower bread crumb firmness and reduced hardening after storage for 2 days at either 20 °C or 7 °C. Considering the results obtained here along with those from the original line, it is clear that the 5-5 genotype can generate specific changes in starch characteristics and staling regardless of its genetic background. Thus, we renamed the 5-5 wheat lines "Slow Staling" (SS) wheat. As expected, our results indicated that the hard wheat SS NIL was more suitable for bread-making. On the other hand, we found that white salted noodle made with the SS NIL of the soft wheat variety had a relatively shorter cooking time, a softer texture, and a reduction in textural changes during storage, all of which are potentially useful for noodle manufacturers.

In hexaploid crops, such as bread wheat, it should be possible to fine-tune phenotypic traits by identifying wild-type and null genes from each of the three genomes and combining them in a calculated manner. Here, we demonstrate this with gene combinations for two starch synthesis genes, SSIIa and GBSSI. Lines with inactive copies of both enzymes show a very dramatic change in phenotype, so to create intermediate phenotypes, we used marker-assisted selection to develop near-isogenic lines (NILs) carrying homozygous combinations of null alleles. For both genes, gene dosage effects follow the order BD ≥ A; therefore, we completed detailed analysis of starch characteristics for NIL 3-3, which is null for the B-genome copy of the SSIIa and GBSSI genes, and NIL 5-5, which has null mutations in the B- and D-genome-encoded copies of both of these genes. The effects of the combinations on phenotypic traits followed the order expected on the basis of genotype, with NIL 5-5 showing the largest differences from the wild type, while NIL 3-3 characteristics were intermediate between NIL 5-5 and the wild type. Differences among genotypes were significant for many starch characteristics, including percent amylose, chain length distribution, gelatinization temperature, retrogradation, and pasting properties, and these differences appeared to translate into improvements in end-product quality, since bread made from type 5-5 flour showed a 3 day lag in staling.

Near-isogenic lines (NILs) of the eight haplotypes of starch synthase IIa (SSIIa) were used to analyze the effects of SSIIa gene dosage on branch chain length, gelatinization, pasting, retrogradation, and enzymatic hydrolysis of starches. Compared to wild-type, the amylopectin of lines missing one or more active SSIIa enzymes had increases in the proportion of short branch chains (DP6-10) and decreases in midlength chains (DP11-24), and the size of these differences depended on the dosage of active SSIIa enzymes. Of the three loci, SSIIa-A1 had the smallest contribution to amylopectin structure and SSIIa-B1 the largest. The different effects of the three SSIIa enzymes on starch properties were also seen in gelatinization, retrogradation, pasting, and enzymatic hydrolysis properties. Such differences in starch properties might be useful in influencing the texture and shelf life of food products.

Sweet wheat (SW), which lacks functional granule-bound starch synthase I (GBSSI) and starch synthase IIa (SSIIa), accumulates high levels of free sugars in immature seeds. Here, we examined the effects of the lack of these two enzymes on mature kernel composition. Whole grain flour of SW had higher levels of sugars, particularly maltose, slightly higher ash and protein content, approximately two to three times higher lipid levels, and about twice as much total dietary fiber as parental or wild-type lines. Considerably higher levels of low-molecular-weight soluble dietary fiber (LMW-SDF), largely consisting of fructan, were also detected in SW. Although there were no differences in total amino acid levels, the free amino acid content of SW was approximately 4-fold higher than that of wild type, and the levels of certain free amino acids such as proline were particularly high. Thus, we were able to clearly demonstrate that the lack of GBSSI and SSIIa caused dramatic changes in mature seed composition in SW. These compositional changes suggest that SW flour may provide health benefits when used as a food ingredient.