a b s t r a c tIn cereal seeds, mutations in one or more starch synthases lead to decreased amylopectin and increased amylose content. Here, the impact of starch synthase IIa (SSIIa or SGP-1) mutations upon durum starch was investigated. A screen of durum accessions identified two lines lacking SGP-A1, the A genome copy of SGP-1. The two lines were determined to carry a 29 bp deletion in the first exon of SSIIa. The SGP-A1 nulls were crossed with the durum variety 'Mountrail' and F 5 derived SGP-A1 null progeny lines were treated with EMS. From each EMS population, one SGP-B1 null mutation was recovered with each being a missense mutation. Each of the SGP-1 nulls was found to have large increases in amylose content and reduced binding of SGP-2 and SGP-3 to the interior of starch granules. RNA-Seq was used to examine the impact the loss of SGP-1 has upon other starch biosynthetic genes. Significant increases in transcript levels of several starch biosynthetic genes were observed in SGP-1 nulls relative to Mountrail. The resultant high amylose durums may prove useful in the creation of value added pasta with increased firmness and reduced glycemic index.
Cereal Chem. 75(1):162-165Granule-bound starch synthase (GBSS) is the primary enzyme responsible for the synthesis of amylose in amyloplasts of cereal endosperm cells. Bread wheats, due to their hexaploid genetic system, carry three genes (wx loci) encoding GBSS. Purification and separation of GBSS from more than 200 North American hexaploid wheats allowed the identification of genotypes that carry null alleles at either the wx-A1 and wx-B1 loci. In addition, the cultivar Ike carried both wx-A1 and wx-B1 null alleles. No wx-D1 nulls were detected. Null alleles were found in 10% of the hard winter wheats tested, but in only 2% of the sampled soft winter wheats. Amylose contents of wheats carrying single null alleles at either the wx-A1 or wx-B1 loci often were lower than those of wild type wheats, but greater reduction in amylose content was observed in Ike. Monoclonal antibodies were used to quantify water-extractable GBSS in both wild-type and null genotypes. Gene dosage compensation was evident, although GBSS content, as measured by ELISA, was significantly lower in Ike than in all other wheats. The identification of null alleles in adapted genotypes suggests the development of wheats with a wide range of amylose contents will be possible by simple genetic crossing and selection.
An automated sorting system was developed that nondestructively measured quality characteristics of individual kernels using near‐infrared (NIR) spectra. This single‐kernel NIR system was applied to sorting wheat (Triticum aestivum L.) kernels by protein content and hardness, and proso millet (Panicum miliaceum L.) into amylose‐bearing and amylose‐free fractions. Single wheat kernels with high protein content could be sorted from pure lines so that the high‐protein content portion was 3.1 percentage points higher than the portion with the low‐protein kernels. Likewise, single wheat kernels with specific hardness indices could be removed from pure lines such that the hardness index in the sorted samples was 29.4 hardness units higher than the soft kernels. The system was able to increase the waxy, or amylose‐free, millet kernels in segregating samples from 94% in the unsorted samples to 98% in the sorted samples. The portion of waxy millet kernels in segregating samples was increased from 32% in the unsorted samples to 55% after sorting. Thus, this technology can be used to enrich the desirable class within segregating populations in breeding programs, to increase the purity of heterogeneous advanced or released lines, or to measure the distribution of quality within samples during the marketing process.
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