BackgroundStarch is stored in higher plants as granules composed of semi-crystalline amylopectin and amorphous amylose. Starch granules provide energy for the plant during dark periods and for germination of seeds and tubers. Dietary starch is also a highly glycemic carbohydrate being degraded to glucose and rapidly absorbed in the small intestine. But a portion of dietary starch, termed “resistant starch” (RS) escapes digestion and reaches the large intestine, where it is fermented by colonic bacteria producing short chain fatty acids (SCFA) which are linked to several health benefits. The RS is preferentially derived from amylose, which can be increased by suppressing amylopectin synthesis by silencing of starch branching enzymes (SBEs). However all the previous works attempting the production of high RS crops resulted in only partly increased amylose-content and/or significant yield loss.ResultsIn this study we invented a new method for silencing of multiple genes. Using a chimeric RNAi hairpin we simultaneously suppressed all genes coding for starch branching enzymes (SBE I, SBE IIa, SBE IIb) in barley (Hordeum vulgare L.), resulting in production of amylose-only starch granules in the endosperm. This trait was segregating 3:1. Amylose-only starch granules were irregularly shaped and showed peculiar thermal properties and crystallinity. Transgenic lines retained high-yield possibly due to a pleiotropic upregualtion of other starch biosynthetic genes compensating the SBEs loss. For gelatinized starch, a very high content of RS (65 %) was observed, which is 2.2-fold higher than control (29%). The amylose-only grains germinated with same frequency as control grains. However, initial growth was delayed in young plants.ConclusionsThis is the first time that pure amylose has been generated with high yield in a living organism. This was achieved by a new method of simultaneous suppression of the entire complement of genes encoding starch branching enzymes. We demonstrate that amylopectin is not essential for starch granule crystallinity and integrity. However the slower initial growth of shoots from amylose-only grains may be due to an important physiological role played by amylopectin ordered crystallinity for rapid starch remobilization explaining the broad conservation in the plant kingdom of the amylopectin structure.
SummaryGrain starch phosphorylation and amylose content affect germination and seedling establishment through the combination of direct effects on altered starch granule and molecular structure and indirect effects on amylase activities.
The importance of cereal starch production worldwide cannot be overrated. However, the qualities and resulting values of existing raw and processed starch do not fully meet future demands for environmentally friendly production of renewable, advanced biomaterials, functional foods, and biomedical additives. New approaches for starch bioengineering are needed. In this review, we discuss cereal starch from a combined universal bioresource point of view. The combination of new biotechniques and clean technology methods can be implemented to replace, for example, chemical modification. The recently released cereal genomes and the exploding advancement in whole genome sequencing now pave the road for identifying new genes to be exploited to generate a multitude of completely new starch functionalities directly in the cereal grain, converting cereal crops to production plants. Newly released genome data from cereal ancestors can potentially allow for the reintroduction of cereal traits including, for example, health‐promoting carbohydrates that may have been lost during domestication. Raw materials produced in this manner can be processed by clean enzyme‐assisted techniques or thermal treatment in combination to further functionalize or stabilize the starch polymers. Importantly, such products can be multifunctional in the sense of combined food/material or food/pharma purposes, for example, edible plastics, shape memory materials, and cycloamylose carriers and stabilizers for diverse bioactives.
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