Cunxu Wei

Microstructure and ultrastructure of high-amylose rice resistant starch granules modified by antisense RNA inhibition of starch branching enzyme.

A high-amylose transgenic rice line (TRS) modified by antisense RNA inhibition of starch branching enzymes revealed a resistant starch-rich quality. Compound starch granules in whole grains of the regular rice cultivar Teqing (TQ) were readily split during fracturing, whereas the starch granules in TRS were structurally intact and showed large voluminous, non-angular rounded bodies and elongated, filamentous structures tolerant of fracturing. In isolated preparation, TQ starch granules broke up into separate polygonal granules, whereas TRS starch granules kept their intactness. TRS starch granules consisted of packed smaller subgranules, some of which located at the periphery of starch granules were fused to each other with adjacent ones forming a thick band or wall encircling the entire circumference of the granules. TQ starch granules had a high concentration of amylose in the concentric hilum, whereas TRS starch granules showed a relatively even distribution of amylose with intense amylose in both hilum and band.

C-Type Starch from High-Amylose Rice Resistant Starch Granules Modified by Antisense RNA Inhibition of Starch Branching Enzyme

High-amylose starch is a source of resistant starch (RS) which has a great benefit on human health. A transgenic rice line (TRS) enriched amylose and RS had been developed by antisense RNA inhibition of starch branching enzymes. In this study, the native starch granules were isolated from TRS grains as well as the wild type, and their crystalline type was carefully investigated before and after acid hydrolysis. In high-amylose TRS rice, the C-type starch, which might result from the combination of both A-type and B-type starch, was observed and subsequently confirmed by multiple physical techniques, including X-ray powder diffraction, solid-state nuclear magnetic resonance, and Fourier transform infrared. Moreover, the change of starch crystalline structure from C- to B-type during acid hydrolysis was also observed in this RS-rich rice. These data could add to our understanding of not only the polymorph structure of cereal starch but also why high-amylose starch is more resistant to digestion.

Granule structure and distribution of allomorphs in C-type high-amylose rice starch granule modified by antisense RNA inhibition of starch branching enzyme.

C-type starch, which is a combination of both A-type and B-type crystal starch, is usually found in legumes and rhizomes. We have developed a high-amylose transgenic line of rice (TRS) by antisense RNA inhibition of starch branching enzymes. The starch in the endosperm of this TRS was identified as typical C-type crystalline starch, but its fine granular structure and allomorph distribution remained unclear. In this study, we conducted morphological and spectroscopic studies on this TRS starch during acid hydrolysis to determine the distribution of A- and B-type allomorphs. The morphology of starch granules after various durations of acid hydrolysis was compared by optical microscopy, scanning electron microscopy, and transmission electron microscopy. The results showed that amorphous regions were located at the center part of TRS starch subgranules. During acid hydrolysis, starch was degraded from the interior of the subgranule to the outer surface, while the peripheral part of the subgranules and the surrounding band of the starch granule were highly resistant to acid hydrolysis. The spectroscopic changes detected by X-ray powder diffraction, 13C cross-polarization magic-angle spinning NMR, and attenuated total reflectance Fourier transform infrared showed that the A-type allomorph was hydrolyzed more rapidly than the B-type, and that the X-ray diffraction profile gradually changed from a native C-type to a CB-type with increasing hydrolysis time. Our results showed that, in TRS starch, the A-type allomorph was located around the amorphous region, and was surrounded by the B-type allomorph located in the peripheral region of the subgranules and the surrounding band of the starch granule. Thus, the positions of A- and B-type allomorphs in the TRS C-type starch granule differ markedly from those in C-type legume and rhizome starch.

Structural and functional properties of C-type starches.

This study investigated the structural and functional properties of C-type starches from pea seeds, faba bean seeds, yam rhizomes and water chestnut corms. These starches were mostly oval in shape with significantly different sizes and contents of amylose, damaged starch and phosphorus. Pea, faba bean and water chestnut starches had central hila, and yam starch had eccentric hilum. Water chestnut and yam starches had higher amylopectin short and long chain, respectively. Water chestnut and faba bean starches showed CA-type crystallinities, and pea and yam starches had C-type crystallinities. Water chestnut starch had the highest swelling power, granule swelling and pasting viscosity, lowest gelatinization temperatures and enthalpy. Faba bean starch had the lowest pasting viscosity, whereas yam starch had the highest gelatinization temperatures. Water chestnut and yam starches possessed significantly higher and lower susceptibility to acid and enzyme hydrolysis, the highest and lowest RDS contents, and the lowest and highest RS contents, respectively.

Structural Changes of High-Amylose Rice Starch Residues following in Vitro and in Vivo Digestion

High-amylose cereal starch has a great benefit on human health through its resistant starch content. In this paper, starches were isolated from mature grains of high-amylose transgenic rice line (TRS) and its wild-type rice cultivar Te-qing (TQ) and digested in vitro and in vivo. The structural changes of digestive starch residues were characterized using DSC, XRD, (13)C CP/MAS NMR, and ATR-FTIR. TQ starch was very susceptible to digestion; its residues following in vitro and in vivo digestion showed similar structural characteristics with TQ control starch, which suggested that both amorphous and crystalline structures were simultaneously digested. Both amorphous and the long-range order structures were also simultaneously hydrolyzed in TRS starch, but the short-range order (double helix) structure in the external region of TRS starch granule increased with increasing digestion time. The A-type polymorph of TRS C-type starch was hydrolyzed more rapidly than the B-type polymorph. These results suggested that B-type crystallinity and short-range order structure in the external region of starch granule made TRS starch resistant to digestion.

Morphology, structure and gelatinization properties of heterogeneous starch granules from high-amylose maize

High-amylose cereal endosperm is rich in heterogeneous starch granules. In this paper, we investigated the morphology, structure and gelatinization properties of high-amylose maize endosperm starch. Starch had individual, aggregate and elongated heterogeneous granules. Most of individual granules were round with small size and had one central hilum. Aggregate and elongated granules consisted of many subgranules with central hila, and had irregular and rod/filamentous shapes, respectively. Iodine stained starch granules showed five types of polarization colors: blue, purple, fuchsia, dark red, and interior dark blue and exterior brown. Most of individual and aggregate granules had the color of dark red, that of elongated granules the color of interior dark blue and exterior brown. Amylose was mainly distributed in the hilum region and the circumference of starch granules. Aggregate and elongated granules had higher amylose content than individual granules. Elongated and individual granules had the highest and the lowest gelatinization resistance among high-amylose maize heterogeneous starch granules, respectively.

Formation of semi-compound C-type starch granule in high-amylose rice developed by antisense RNA inhibition of starch-branching enzyme.

Cereal starch granules with high-amylose and resistant starch (RS) always show irregular morphology and special crystalline structure, but their formation during grain development is not yet clear. In our previous studies, we had generated a transgenic rice line (TRS) enriched with amylose and RS, which contained semi-compound starch showing a C-type crystalline structure. In this study, the formation of semi-compound C-type starch granule during TRS endosperm development was carefully investigated with light, scanning electron, and transmission electron microscopes and X-ray powder diffraction. The results showed that the TRS starch subgranules, each with a central hilum, were individually initiated in amyloplast and showed an A-type crystal at the early stage of starch granule development, which was similar to that in its wild type. However, with the endosperm development, the amylose content in TRS endosperm starch increased and the B-type starch crystal was deposited in the periphery of subgranules; then, the adjacent subgranules fused together and finally formed a continuous outer layer band surrounding the entire circumference of the starch granule. Accordingly, a mechanistic model for the formation of semi-compound C-type starch granules is proposed.

Structural and functional properties of alkali-treated high-amylose rice starch

Native starches were isolated from mature grains of high-amylose transgenic rice TRS and its wild-type rice TQ and treated with 0.1% and 0.4% NaOH for 7 and 14 days at 35 °C. Alkali-treated starches were characterised for structural and functional properties using various physical methods. The 0.1% NaOH treatment had no significant effect on structural and functional properties of starches except that it markedly increased the hydrolysis of starch by amylolytic enzymes. The 0.4% NaOH treatment resulted in some changes in structural and functional properties of starches. The alkali treatment affected granule morphology and decreased the electron density between crystalline and amorphous lamellae of starch. The effect of alkali on the crystalline structure including long- and short-range ordered structure was not pronounced. Compared with control starch, alkali-treated TRS starches had lower amylose content, higher onset and peak gelatinisation temperatures, and faster hydrolysis of starch by HCl and amylolytic enzymes.

Different structural properties of high-amylose maize starch fractions varying in granule size.

Large-, medium-, and small-sized granules were separated from normal and high-amylose maize starches using a glycerol centrifugation method. The different-sized fractions of normal maize starch showed similar molecular weight distribution, crystal structure, long- and short-range ordered structure, and lamellar structure of starch, but the different-sized fractions of high-amylose maize starch showed markedly different structural properties. The amylose content, iodine blue value, amylopectin long branch-chain, and IR ratio of 1045/1022 cm(-1) significantly increased with decrease of granule size, but the amylopectin short branch-chain and branching degree, relative crystallinity, IR ratio of 1022/995 cm(-1), and peak intensity of lamellar structure markedly decreased with decrease of granule size for high-amylose maize starch. The large-sized granules of high-amylose maize starch were A-type crystallinity, native and medium-sized granules of high-amylose maize starch were CA-type crystallinity, and small-sized granules of high-amylose maize starch were C-type crystallinity, indicating that C-type starch might contain A-type starch granules.

Different structures of heterogeneous starch granules from high-amylose rice.

High-amylose cereal starches usually have heterogeneous starch granules in morphological structure. In the present study, the polygonal, aggregate, elongated, and hollow starch granules were separated from different regions of the kernels of high-amylose rice, and their structures were investigated. The results showed that the polygonal starch granules had low amylose content and high short branch-chain and branching degree of amylopectin, and exhibited A-type crystallinity. The aggregate starch granules had high long branch-chain of amylopectin, relative crystallinity, and double helix content, and exhibited C-type crystallinity. The elongated starch granules had high amylose content and low branching degree of amylopectin and relative crystallinity, and exhibited C-type crystallinity. The hollow starch granules had very high amylose content, proportion of amorphous conformation, and amylose-lipid complex, and very low branch-chain of amylopectin, branching degree of amylopectin, and double helix content, and exhibited no crystallinity. The different structures of heterogeneous starch granules from high-amylose rice resulted in significantly different thermal properties.