Starch Fine Structure Research Articles

Effects of Wx, SSIIa and FLO2 alleles and their interactions on the formation of multi-scale structures of rice starch.

The multi-scale structures of rice starches from 16 recombinant inbred lines and their parents with different combinations of Wx, SSIIa and FLO2 alleles were investigated. Results indicated that the starch multi-scale structures were significantly affected by genotype, environment and genotype × environment interaction. Wx, SSIIa and FLO2 alleles and their interactions significantly affected the starch fine structure. Starches with Wxa allele had higher amylose content (AC) in different amylose chain regions (hAm,i~iii), degree of polymerization (DP) of amylose peak (XAm), Bragg long period distance (dac), but lower relative crystallinity (RC) compared with Wxb. The starches with SSIIaGC allele had higher XAm, more amylopectin B1 chains (fb1) and RC, but less A chains (fa) and dac than those of SSIIaTT. The floury2 (flo2) mutant starch displayed increased ratio of the peak heights of amylopectin (hAP2/hAP1), fb2 and dac, but decreased RC compared to the wildtype. The Wx × SSIIa interaction had a significant impact on AC, XAP1, βAm,iii, average chain length of amylopectin molecules (X¯) and RC. The interaction effects of FLO2 × Wx were significant for AC, hAm/hAp1, hAm,ii, βAm,iii, hAm,iii, RC, D and dac. Significant FLO2 × SSIIa interaction was detected for XAM, hAP2/hAP1, βAm,ii, fb1, fb3 and RC. FLO2 × Wx×SSIIa effect was significant for XAP1, hAP2/hAP1, βAm,ii, fa, fb3 and X¯. This study will help us to further understand the individual and interaction effects of the Wx, SSIIa and FLO2 and the effect of environments on the starch fine structure, but also promote high-quality rice breeding through structural modification of starch.

Modulating the assembly of starch-lecithin inclusion complexes and its underlying mechanism for controlling starch digestibility under the dense system

Starch-lipid inclusion complexes were used to regulate starch digestibility by changing the binding affinity of digestive enzymes such as α-amylase and α-glucosidase. However, when above a critical aggregation concentration (CAC) of starch and lipid molecules, they tended to self-assemble into double helices or normal micelles respectively, rather than interacting with each other to form starch-lipid complexes. Thus, how to enhance the complexation efficiency under a dense system is the key approach for the improvement of starch bioaccessibility and nutritional function. Herein, based on the dimethyl sulfoxide (DMSO) reverse solvent method, the effect of water/DMSO ratios and starch chain structures on the interaction and assembly behaviors between starch and lecithin molecules in high concentrations were systematically investigated. The results revealed that water/DMSO in a ratio of 1.5:1 promoted the interactions between cavities of starch helices and long alkyl chains of lecithin molecules, with higher level of complexed lecithin content, relative crystallinity (RC), enthalpy (ΔH) (Tp＞100°C), ordered-aggregation structures and resistant starch (RS) content. In this kind of solvent, starch samples with higher content of amylose and longer chains (degree of polymerization (DP) of B2 chains and B3+ chains) is advantageous to the formation of VII-type inclusion starch-lecithin complexes, with a rise in RS from 14.17% to 43.64%. It provided an effective method to modulate starch digestibility under the dense system of starch and lecithin via modulating the water/DMSO ratios and starch fine structures and lay the foundation for the large-scale preparation and application of starch-lecithin complexes.

Physicochemical Properties and Fine Structure of Starch in Jinong Xiangruan 1 and DGR1 Soft Rice Varieties Cultivated in Different Regions of China

Rice, a staple food for billions around the globe, is cultivated in numerous forms. Among them, soft rice is well known, which is characterized by its tender, creamy consistency and desirable texture. In this study, we examined the physicochemical properties and fine structure of starch from two soft rice varieties, Jinong Xiangruan 1 and DGR1, cultivated in different regions in China (Baodi District, Tianjin City; Liaoning Province; and Fengyang City, Anhui Province). The aim was to understand how amylopectin content (AC) influences rice quality. This research aims to bridge the knowledge gap regarding the role of amylopectin in determining rice’s adhesive consistency and viscosity. Significant regional differences were observed in yield components such as the number of grains per panicle, seed setting rates and 1000-grain weight, with Liaoning generally showing higher performance metrics compared to other regions. Physicochemical analysis highlighted that though glue consistency and taste values showed little regional variation, AC significantly influenced rice hardness and viscosity. Rapid Visco Analyzer (RVA) profile analysis further demonstrated distinct differences in viscosity characteristics, underscoring the regional impacts on starch behavior. Additionally, molecular weight distribution and amylopectin chain length analysis, conducted via SEC-MALLS-RI and ICS ion chromatography, revealed notable differences in starch composition across varieties and locations. The findings suggest that environmental conditions play a crucial role in defining starch characteristics and, consequently, the eating quality of rice. This provides valuable insights for breeding high-quality japonica rice with broad adaptability.

Relationship between Chalkiness and the Structural and Physicochemical Properties of Rice Starch at Different Nighttime Temperatures during the Early Grain-Filling Stage.

The chalkiness, starch fine structure, and physiochemical properties of rice starch were analyzed and their correlations were investigated under different nighttime temperatures during the early grain-filling stage. Compared to MT, medium temperature (MT) and low (LNT) and high (HNT) nighttime temperatures resulted in an increased chalky grain rate (CGR) and chalkiness degree (CD). LNT mainly affected the chalkiness by increasing peak1 (short branch chains of amylopectin), the branching degree, and the proportion of small starch granules but decreasing peak2 (long branch chains of amylopectin) and peak3 (amylose branches). This altered the pasting properties, such as by increasing the peak viscosity and final viscosity. However, HNT mainly affected the chalkiness by increasing peak2 and the crystallinity degree but decreasing peak1 and peak3. Regarding the thermal properties, HNT also elevated peak and conclusion temperatures. The CGR and CD were significantly and positively correlated with the proportions of small and medium starch granules, peak1, branching degree, gelatinization enthalpy, setback viscosity, and pasting time but markedly and negatively correlated with the proportion of large starch granules, amylose content, peak3, peak viscosity, and breakdown viscosity. These findings suggest that LNT and HNT disrupted the starch structure, resulting in increased chalkiness. However, their mechanisms of action differ.

Rheological properties of indica rice determined by starch structure related enzymatic activities during after-ripening

The processing quality of indica rice must undergo ripening after harvest to achieve stability and improvement. However, the mechanism underlying this process remains incompletely elucidated. Starch, the predominant component in indica rice, plays a crucial role in determining its properties. This study focused on analyzing the rheological properties and starch fine structure, as well as the related biosynthetic enzymes of indica rice during the after-ripening process. The results showed that after-ripened rice exhibited increased elastic modulus (G′) and viscous modulus (G″), accompanied by a decrease in the loss tangent (Tan δ), indicating an enhancement in viscoelasticity and the gel network structure. Moreover, the proportions of amylopectin super long chains (DP 37–60) decreased, while those of medium chains (DP 13–24 and DP 25–36) or short chains (DP 6–12) of amylopectin increased. Additionally, the activities of starch branching enzyme (SBE) and starch debranching enzyme (DBE) declined over the after-ripening period. Pearson correlation analysis revealed that the rheological properties of after-ripened rice were correlated with the chain length distribution (CLD) of starch, which, in turn, was associated with its related endogenous enzymes. These findings provied new insights into understanding the quality changes of after-ripened indica rice.

Amylopectin chain length distributions and amylose content are determinants of viscoelasticity and digestibility differences in mung bean starch and proso millet starch

This study aimed to reveal the underlying mechanisms of the differences in viscoelasticity and digestibility between mung bean starch (MBS) and proso millet starch (PMS) from the viewpoint of starch fine molecular structure. The contents of amylopectin B2 chains (14.94–15.09 %), amylopectin B3 chains (14.48–15.07 %) and amylose long chains (183.55–198.84) in MBS were significantly higher than PMS (10.45–10.76 %, 12.48–14.07 % and 70.59–88.03, respectively). MBS with higher amylose content (AC, 28.45–31.80 %) not only exhibited a lower weight-average molar mass (91,750.65–128,120.44 kDa) and R1047/1022 (1.1520–1.1904), but also was significantly lower than PMS in relative crystallinity (15.22–23.18 %, p < 0.05). MBS displayed a higher storage modulus (G′) and loss modulus (G′′) than PMS. Although only MBS-1 showed two distinct and discontinuous phases, MBS exhibited a higher resistant starch (RS) content than PMS (31.63–39.23 %), with MBS-3 having the highest RS content (56.15 %). Correlation analysis suggested that the amylopectin chain length distributions and AC played an important role in affecting the crystal structure, viscoelastic properties and in vitro starch digestibility of MBS and PMS. These results will provide a theoretical and scientific basis for the development of starch science and industrial production of low glycemic index starchy food.

Investigating the evolution of the fine structure in cassava starch during growth and its correlation with gelatinization performance

The evolution of the starch fine structure during growth and its impact on the gelatinization behavior of cassava starch (CS) was investigated by isolating starch from South China 6068 (SC6068) cassava harvested from the 4th to 9th growth period. During growth, the short-range ordered structure, crystallinity as well as particle size distribution of starch were increased. Meanwhile, the starch molecular size and amylopectin (AP) proportion increased, while the proportion of amylose (AM) exhibited a decreasing tendency. The chains of short-AM (X ~ 100–1000) were mainly significantly reduced, whereas the short and medium-AP chains (X ~ 6–24) had the most increment in AP. The solubility, thermal stability, shear resistance, and retrogradation resistance of starch were enhanced after gelatinized under the influence of the results mentioned above. This study presented a deeper insight into the variation of starch fine structure during growth and its influence on gelatinization behavior, which would provide a theoretical basis for starch industrial applications.

Enhancing in vitro starch digestion of tartary buckwheat: Unveiling the vital role of endogenous polyphenols in starch fine structure

In this study, the effect of buckwheat polyphenols (rutin and quercetin) on in vitro digestion of cooked starch was analyzed from view of the grain source, fine structure of starch and enzymatic inhibition. The results showed no significant difference in the inhibition of rutin on tartary buckwheat starch digestion as compared with common buckwheat and wheat starches. However, their inhibitory effects were negatively correlated with the content of long amylose chains (2000 < DP < 20000) but positively correlated with that of short amylose chains (100 < DP < 500) and amylopectin (13 < DP < 24) chains (p < 0.05). There was a significant difference in the digestive inhibition of rutin on starches obtained from different tartary buckwheat varieties, and the optimal ratio of rutin (2%) addition for TS1 (tartary buckwheat starch, Heifeng No.1). Quercetin showed stronger inhibition on the digestion of tartary buckwheat starch, which was attributed to the following fact: quercetin can significantly change the structure of enzymatic molecular, thereby inhibiting enzyme activity. Overall, the inhibition of rutin on starch digestion was mainly determined by starch fine structure, and 2% of rutin could show optimal inhibition on the digestion of tartary buckwheat starch.

A comparative study of grain quality and physicochemical properties of premium japonica rice from three typical production regions.

Grain quality indicates rice commodity value. This research compared grain quality and physicochemical properties of premium japonica rice from three production regions, Yangtze River downstream of China (YRDCN), Northeast region of China (NECN) and Japan. Results showed that there were distinct quality and physicochemical characteristics variance among the three groups of japonica rice, while CVs of most quality parameters from low to high was Japan, YRDCN and NECN. YRDCN rice presented obvious lower apparent amylose content (AAC) and ratio of each chain-length sections of amylopectin, and showed higher protein contents especially glutelin and ratio in short and intermediate amylopectin molecules. Among three rice groups, YRDCN rice presented weaker appearance, whereas did not show inferior cooking and eating properties, which was primarily linked to lower AAC. Rice AAC and starch fine structure significantly correlated with pasting parameters, swelling power and solubility, while protein content had close relation with taste analyzer parameters. Results of this study indicated improvement direction for japonica rice of YRDCN, and also provided reference for consumers' rice purchasing selection in accordance with individual taste preference.

The important role of starch fine molecular structures in starch gelatinization property with addition of sugars/sugar alcohols

The relationship between the fine structure of starch and its gelatinization properties is not well studied, particularly in relation to the influence of sugar or sugar alcohol. In this study, seven starches with distinct molecular structures were investigated to determine how different sugars and sugar alcohols affect their gelatinization properties. The inclusion of sugars and sugar alcohols resulted in a significant elevation of starch gelatinization temperatures (∼ 8 °C), especially with sucrose, isomaltose and isomalt. Nevertheless, the influence of these sugars/ sugar alcohols on the gelatinization temperature range and enthalpy change varied depending on the particular starch varieties. According to the correlation analysis, sugars and sugar alcohols mainly exert their impact on the starch gelatinization temperature range and enthalpy change by possibly interacting with amylose chains possessing a degree of polymerization ranging from 100 to 1000 (p < 0.05) and inhibiting the amylose leaching during gelatinization. These findings help a better understanding of the complex relationship between starch fine structure and gelatinization properties under the influence of sugars and sugar alcohols.

Elevated Ripening Temperature Mitigates the Eating Quality Deterioration of Rice in the Lower Grain Position Due to the Improvement of Starch Fine Structure and Properties

Elevated ripening temperature (ET) impacts rice grain quality. In this study, two rice varieties were investigated to evaluate the characterization of starch fine structure and grain eating quality under ET conditions. Rice exposure to ET increased the proportion of large-sized granules and starch granule average size, regardless of grain position. Compared to normal temperature (NT), the amylose content (AM) in the upper grain position (UP) exhibited a significant increase under ET, whereas the contrary results showed a decrease in the lower grain position (LP), and the proportion of shorter amylopectin chains increased under ET in UP or LP, whereas the proportion of long amylopectin chains decreased, resulting in a higher starch gelatinization temperature and enthalpy under ET. For grain position, compared to LP, UP had higher AM and a higher proportion of long amylopectin chains, leading to higher gelatinization enthalpy under ET. For eating quality, we found that ET deteriorated the eating quality of rice compared to NT, and UP had higher eating quality than LP under NT; however, there was a comparable eating quality between UP and LP under ET. Hence, elevated grain filling temperature mitigated the eating quality deterioration of rice grain in LP due to the lower AM and gelatinization enthalpy and the higher proportion of shorter amylopectin chains.

Effects of Starch Molecular Structure and Physicochemical Properties on Eating Quality of Indica Rice with Similar Apparent Amylose and Protein Contents.

It is important to clarify the effects of starch fine structure and protein components on the eating quality of indica rice. In this study, seven indica rice varieties with similar apparent amylose content (AAC) and protein content (PC) but different sensory taste values were selected and compared systematically. It was found that except for AAC and PC, these varieties showed significant differences in starch molecular structure and protein components. Compared with rice varieties with a low sensory taste value, varieties with a higher sensory taste value showed significantly lower amylose and higher amylopectin short chains (degree of polymerization 6-12) content; the protein component showed that the varieties with good taste value had higher albumin and lower globulin and glutelin content (p < 0.05). Rice varieties with lower AC, globulin, and glutelin content, as well as a higher content of albumin and amylopectin short chains, resulted in a higher swelling factor, peak viscosity, breakdown value, and ratio of hardness to stickiness, in which condition cooked rice showed a higher sensory taste value. Moreover, this study indicated that rice varieties with a higher content of albumin and amylopectin short chains were conducive to the good appearance of cooked rice. This study lays the foundation for the taste evaluation of good-tasting indica rice.

Creating high-resistant starch rice by simultaneous editing of SS3a and SS3b.

Creating high-resistant starch rice by simultaneous editing of SS3a and SS3b.

Combined effects of SSII-2RNAi and different Wx alleles on rice grain transparency and physicochemical properties

Near-isogenic lines Nip(Wxb/SSII-2), Nip(Wxb/ss2-2), Nip(Wxmw/SSII-2), Nip(Wxmw/ss2-2), Nip(Wxmp/SSII-2) and Nip(Wxmp/ss2-2) in the Nipponbare (Nip) background containing the SSII-2RNAi cassette combined with different Waxy (Wx) alleles were investigated in terms of rice grain transparency and quality profiles. Rice lines carrying the SSII-2RNAi cassette displayed downregulation of SSII-2, SSII-3 and Wx genes. Introduction of the SSII-2RNAi cassette decreased apparent amylose content (AAC) in all transgenic lines, but grain transparency differed between low AAC rice lines. Grains from Nip(Wxb/SSII-2) and Nip(Wxb/ss2-2) were transparent, while those of rice were increasingly translucent with decreasing moisture due to cavities within starch granules. Rice grain transparency was positively correlated with grain moisture and AAC, but negatively correlated with cavity area within starch granules. Starch fine structure analysis revealed a marked increase in short amylopectin chains with DP 6–12, but a decrease in intermediate chains with DP 13–24, resulting in decreased gelatinisation temperature. Starch crystalline structure analysis showed that the transgenic rice starches have lower crystallinity and lamellar repeat distance than controls due to differences in starch fine structure. The results highlight the molecular basis underpinning rice grain transparency, and provide strategies for improving rice grain transparency.

Subtle differences in starch fine molecular structure are associated with large differences in texture and digestibility of Chinese steamed bread

Chinese steamed bread (CSB) is a staple food for a large number of people in China, but it has a high glycemic index, which is nutritionally undesirable. Developing a CSB with more slowly digestible starch has been achieved, but current processes for this result in a product with poor sensory appeal. In this study, the relations among starch chain-length distribution, starch digestibility and texture of CSBs made from 10 different wheat flour varieties were investigated, the long-term aim being to develop CSBs with an optimal combination of slow starch digestibility and desirable texture. Although there were only small differences among the CLDs of different wheat flours, they were strongly correlated to the differences of both starch digestibility and textural attributes of CSB. A slower starch digestion rate was associated with relatively longer amylose intermediate to long chains, lower amounts of amylopectin short chains and higher amounts of amylose short chains. A softer CSB texture was related to a relatively higher amount of amylopectin long chains and shorter amylopectin short chains. A higher springiness/resilience of CSB was associated with more amylopectin long chains, longer amylose intermediate to long chains, fewer amylose long chains, and smaller average chain length of amylopectin short chains. A higher cohesiveness of CSB was developed with a higher amount of amylopectin long chains. These results suggest that it is feasible to produce CSB with both slow starch digestibility and desirable texture, by choosing the wheat flour with optimized starch structures. Small variance of wheat starch chain-length distribution can cause a significant difference of starch digestibility and texture of Chinese steamed bread. • Small differences exist among wheat starch chain-length distributions. • Slower starch digestibility associated with higher amounts of amylose short chains. • A softer texture of Chinese steamed bread related to more amylopectin long chains. • Higher springiness of Chinese steamed bread related to fewer amylose long chains.

Combined Effects of BEIIb and SSIIa Alleles on Amylose Contents, Starch Fine Structures and Physicochemical Properties of Indica Rice

Starch branching enzyme IIb (BEIIb) and soluble starch synthase IIa (SSIIa) play important roles in starch biosynthesis in cereals. Deficiency in the BEIIb gene produces the amylose extender (ae) mutant rice strain with increased amylose content (AC) and changes in the amylopectin structure. The SSIIa gene is responsible for the genetic control of gelatinization temperature (GT). The combined effects of BEIIb and SSIIa alleles on the AC, fine structures, and physicochemical properties of starches from 12 rice accessions including 10 recombinant inbred lines (RIL) and their two parents were examined in this study. Under the active BEIIb background, starches with the SSIIa-GC allele showed a higher GT than those with the SSIIa-TT allele, resulting from a lower proportion of A chain and a larger proportion of B1 chains in the amylopectin of SSIIa-GC. However, starch with the BEIIb mutant allele (be2b) in combination with any SSIIa genotype displayed more amylose long chains, higher amylose content, B2 and B3 chains, and molecular order, but smaller relative crystallinity and proportion of amylopectin A and B1 chains than those with BEIIb, leading to a higher GT and lower paste viscosities. These results suggest that BEIIb is more important in determining the structural and physicochemical properties than SSIIa. These results provide additional insights into the structure-function relationship in indica rice rather than that in japonica rice and are useful for breeding rice with high amylose content and high resistant starch.

Identification of a new allele of soluble starch synthase IIIa involved in the elongation of amylopectin long chains in a chalky rice mutant

A chalky endosperm mutant (GM03) induced from an indica rice GLA4 was used to investigate the functional gene in starch biosynthesis. Bulked segregant analysis and sanger sequencing determined that a novel mutation in soluble starch synthase IIIa (SSIIIa) is responsible for the chalky phenotype in GM03. Complementary test by transforming the active SSIIIa gene driven by its native promoter to GM03 recovered the phenotype to its wildtype. The expression of SSIIIa was significantly decreased, while SSIIIa protein was not detected in GM03. The mutation of SSIIIa led to increased expression of most of starch synthesis related genes and elevated the levels of most of proteins in GM03. The CRISPR/Cas9 technology was used for targeted disruption of SSIIIa, and the mutant lines exhibited chalky endosperm which phenocopied the GM03. Additionally, the starch fine structure in the knockout mutant lines ss3a-1 and ss3a-2 was similar with the GM03, which showed increased amylose content, higher proportions of B1 and B2 chains, much lower proportions of B3 chains and decreased degree of crystallinity, leading to altered thermal properties with lower gelatinization temperature and enthalpy. Collectively, these results suggested that SSIIIa plays an important role in starch synthesis by elongating amylopectin long chains in rice.

Low temperature and light combined stress after heading on starch fine structure and physicochemical properties of late-season indica rice with different grain quality in southern China

Late-season indica rice frequently encounters low temperature (LT) along with low light (LL) after heading in southern China, which deteriorates the grain quality by altering starch quality. However, the detailed effects on starch properties of these stressors remain unclear. Herein, two indica rice cultivars with good and poor grain quality were grown under control (CK), LT, and LT + LL conditions after heading and the structural and physicochemical properties of their starch were evaluated. Compared with CK, LT and LT + LL worsened thermal and pasting properties of starch in the two cultivars, mainly because they increased branch chain branching and A chain (DP ≤12), and decreased average branch chain length and crystallinity. Compared with LT, LT + LL deteriorated the pasting properties of the poor-quality cultivar, such as reducing breakdown (BD), final and peak viscosity, which mainly owing to decreasing the starch branching and crystallinity degrees, and increasing the small starch granules (d < 10 μm). Gelatinization enthalpy and BD both had significant and positive correlations with amylose content, the ratio of amylose and amylopectin, B3 chain and crystallinity. Taken together, these results suggest that LT and LT + LL during grain filling can deteriorate the physicochemical properties of starch in late-season indica rice cultivars by disrupting starch multilevel structure, especially upon LT + LL. In particular, while poor-quality cultivar had poorer physicochemical properties, the good-quality cultivar had poorer thermal properties under LT + LL.

Relationship between starch fine structure and simulated oral processing of cooked japonica rice.

Simulated oral processing can be used to evaluate the palatability of cooked rice. Previously, we established a simulated oral processing method using a texture analyzer equipped with a multiple extrusion cell probe (TA/MEC). However, the relationship between oral processing and starch fine structure remains unknown. In this study, we analyzed the oral processing properties using TA/MEC and characterized the starch fine structure of japonica rice by size-exclusion chromatography (SEC) and fluorophore-assisted capillary electrophoresis (FACE). The relationship between starch fine structure and oral processing of cooked japonica rice was further investigated. Cooked rice structure contains fast-breakdown (Type I structure), slow-breakdown (Type II structure) and unbreakable structures (Type III structure). Fast-breakdown and slow-breakdown structure were positively correlated with the content of amylose and shorter amylopectin branches. The content of longer amylopectin branches was positively correlated with the contribution of unbreakable structure. The results indicated that cooked japonica rice varieties with more amylose and shorter amylopectin branches tend to form a harder texture and need more work to break down the fast and slow breakdown structures related to rice kernel fragmentation. Meanwhile, cooked japonica rice varieties possess stronger molecular entanglements due to their longer amylopectin branches and contribute more to the breakdown of unbreakable structures. These results can guide breeders to select rice varieties with desirable eating qualities for cultivation.

Effects of storage on the starch fine structure and physicochemical properties of different rice variety types

The genotype of rice variety is important factor that affect grain quality during storage. In this study, japonica, indica, and indica–japonica hybrid rice were selected to reveal the changes in amylase activities, starch fine structure, and physicochemical properties after one year of storage. Amylase activities decreased after storage and starch molecules were degraded, which, respectively, decreased and increased the content of the amylopectin long chains (DP 37+) and amylose. Changes in the starch fine structure resulted in lower swelling factors and a higher gelatinization temperature. Compared with japonica and indica–japonica hybrid rice, indica rice had a more stable starch fine structure and physicochemical properties after storage. The information obtained from this study provides a better understanding of the changes that occur in starch at the molecular level during storage and will provide insight into the breeding of storable rice varieties.