Granule‐bound SSIIa Protein Content and its Relationship with Amylopectin Structure and Gelatinization Temperature of Rice Starch

Starch characteristics such as gelatinization properties determine the quality of various products of rice. The eating, cooking and processing qualities of rice are a function of the gelatinization of rice starch. The gelatinization temperature (GT) of rice is controlled by the starch synthase IIa (SSIIa) gene. In the present study, the amount of the starch‐associated SSIIa protein and its relationship with amylopectin structure and GT were investigated. The results indicated that the starch associated SSIIa protein content was positively correlated with GT and level of amylopectin chains with degree of polymerization 12‐24 (DP12‐24), and negatively correlated with the fraction of chains with DP6‐11 and short chain ratio (SCR), i.e. DP6‐11/DP12‐24 (P<0.001). GT was also negatively correlated with DP6‐11 SCR, and positively correlated with DP12‐24. The SSIIa protein content and amylopectin structure showed significant difference between the two SSIIa single nucleotide polymorphisms (SNPs, i.e. guanine‐cytosine/thymine‐thymine (GC/TT)) groups. Rice with GC and TT SNPs had average SSIIa protein contents of 1.076 and 0.681, respectively. Rice with the GC SNPs displayed a lower amount of chains with DP6‐11 and more chains with DP 12‐24 than those with TT SNPs. A mechanism is postulated to explain how SSIIa could determine the distribution of amylopectin side‐chains, which affects the gelatinization temperature of rice starch.     

Chain Length Distribution of Amylopectin and Physicochemical Properties of Starch in Myanmar Rice Cultivars

Three different types of amylopectin: long chain length of amylopectin (L-type), medium chain length of amylopectin (M-type), and short chain length of amylopectin (S-type) were classified in 16 rice cultivars (3 for L-type, 6 for M-type, and 7 for S-type amylopectin) based on chain length distribution of amylopectin by fluorophore-assisted carbohydrate electrophoresis through capillary electrophoresis. Frequency distribution of amylopectin chain ratio and short chain ratio supported three types of amylopectin. Ta Soke Pwa and Thet Nu Saba Net Pyan had high values of transition temperatures in a range from (70.7, 74.0, and 77.6°C) to (71.4, 75.0, and 78.8°C) in differential scanning calorimeter analysis. Alkali spreading values and starch gelatinization characteristics in 4 M urea solution showed the association between chain length distributions of amylopectin in Myanmar rice cultivars. A simple linear regression in Myanmar rice cultivars was observed for fa, fb1, amylopectin chain ratio, and short chain ratio with alkali spreading values, with R² values of 0.68, 0.84, 0.84, and 0.81, respectively.     

Waxy Gene Factor and Residual Protein of Rice Starch Granules

Residual protein of rice starch granules prepared from milled rice by protein extraction with sodium dodecylbenzene sulfonate or alkaline protease treatment was related to amylose content. SDS-polyacrylamide disc gel electrophoresis confirmed that the major protein in nonwaxy starch granules was the Wx gene product with subunit molecular weight of about 60,000 daltons. The Wx gene product was absent in waxy starch granules.     

The Influence of Environment on Starch Content and Amylose to Amylopectin Ratio in Wheat

There is evidence that starch content plays an active role in determining dough rheological characteristics. The aim of this study was to determine the influence of environment on starch content and amylose: amylopectin ratio, and how this affects breadmaking quality. Ten hard red spring wheat cultivars were planted in three different environments in a randomized complete block design with three replications. Total starch content, amylose: amylopectin ratio, milling, rheological and baking characteristics were measured. Starch content was significantly influenced by the environment. It was significantly negatively correlated with loaf volume, wet gluten content and flour protein content across the three environments. Starch content and protein content were significantly negatively correlated, yet the value was relatively small (r = −0.4) and the relationship was therefore not directly inverse, and other factors influenced this relationship. Some cultivars interacted with specific environments, and they ranked high for starch content and loaf volume. Amylose: amylopectin ratio was very consistent for the three environments but was not significantly correlated with breadmaking quality characteristics across the environments, although there were significant correlations at individual environments. Cultivar choice and environment where cultivars are planted will therefore affect the starch content. High starch content will not necessarily lead to poor baking quality.     

The Influences of Chain Length of Amylopectin on Resistant Starch in Rice (Oryza sativa L.)

Amylopectin is the principle component of starch. To elucidate the relationships between amylopectin and resistant starch content, six rice mutants with altered fine structure of amylopectin were selected for comparative studies with the primary wild type and two types of amylose‐extender (ae) mutants. Significant differences in resistant starch content were observed among mutants with similarity or differences in amylose levels. Mutants high in resistant starch had significantly increased proportions of short amylopectin chains with DP≤12, decreased levels of intermediate chains with size of 13≤DP≤36, and decreased fractions of long chains with DP≥37. Additionally, there was a mutant different to ae, which was characterized by an increased level of short chains with 8≤DP≤12 and 13≤DP≤24, and a decreased proportion of long chains with DP≥37. The increased contents of short chains with 8≤DP≤12 and decreased of intermediate and long chains with 24≤DP were clearly associated with the increase of resistant starch in rice.     

直支链淀粉对白酒生产的影响

本文比较了不同直支链淀粉含量的大米、高粱的糊化情况和蒸煮香气,并进行了实验室和窖池发酵试验。结果发现酿酒微生物代谢出的酶既能水解支链淀粉也能水解直链淀粉;同一品种的粮食直链淀粉越高,淀粉结构越紧密,糊化时间越长,粮香越浓;不同品种的粮食淀粉颗粒大小、结构不同,糊化时间也不同。不管是直链淀粉还是支链淀粉,只要糊化得好,出酒率差别不大。     

A Study of the Internal Structure in Cassava and Rice Amylopectin

Cassava and rice amylopectins and their ϕ,β‐limit dextrins were debranched and the unit chain compositions were analyzed by gel permeation chromatography and high performance anion‐exchange chromatography. The content of amylose and super‐long amylopectin chains was 15.5‐15.6% and 0.8‐1.2% in cassava and 15.1‐18.7% and 3.4‐8.6% in rice starches, respectively. The internal unit chain profiles of the amylopectin samples were obtained from the ϕ,β‐limit dextrins. Short B‐chains were subdivided into a major group (BS_major) with a degree of polymerization (DP) of 8‐25 and a minor group of “fingerprint” B‐chains (B_fp) with DP 3‐7. Cassava and rice amylopectins possessed different amounts and patterns of B_fp‐chains. The rice samples contained higher amounts of B_fp‐chains, which suggested larger size of its clusters when compared with cassava. The content of the shortest “fingerprint” A‐chains (A_fp) was also variable, especially between the rice cultivars. The position of the internal chains in the original amylopectin chain profile was traced. The reconstructed B‐chain profiles fitted well with the original profiles. This implied that most of the B‐chains carried external chain segments of similar lengths, which corresponded to the experimentally measured average length.     

Amylopectin structure of heat–moisture treated starches

The effects of heat–moisture treatment (HMT; moisture content of 25%, at 100°C for 24 h) on starch chain distribution and unit chain distribution of amylopectin in normal rice, waxy rice, normal corn, waxy corn, normal potato, and waxy potato starches were investigated. After HMT, starch chain distribution (amylose and amylopectin responses) of waxy corn and potato starches were identical to those of untreated starches, whereas the chromatographic response of waxy rice starch showed a slight decrease, but with a slight increase in peak tailing. This result indicated that HMT had no (or very limited) effect on the degradation of amylopectins. Analysis of unit chain distribution of amylopectins revealed that waxy characteristics affected the molecular structure of amylopectin in untreated starches, i.e., the CL of normal‐type starches was greater than that of waxy‐type starches. After HMT, the CL and unit chain distribution of all starches were no different than those of untreated starches. The results implied that changes in the physico‐chemical properties of HMT starches would be due to other phenomena rather than the degradation of amylopectin molecular structure. However, the thermal degradation of amylopectin molecules of waxy starches could occur by HMT at higher treatment temperatures (120 and 140°C).     

Extrusion of Rice,Bean and Corn Starches: Extrudate Structure and Molecular Changes in Amylose and Amylopectin

The objective of this study was to evaluate the effects of starch source and amylose content on the expansion ratio, density, and texture of expanded extrudates, as well as to investigate the structural and molecular changes that occur in starch granules as a function of extrusion. The starches employed were rice starches (8%, 20%, and 32% amylose), carioca bean starch (35% amylose), and Hylon V^® corn starch (55% amylose). The extrudates from rice starches containing 20% and 32% amylose exhibited the highest expansion ratio, while, extrudates from Hylon V^® corn starch containing 55% amylose exhibited the lowest expansion ratio. The hardness values of the extrudates with 55% amylose were twice those of the extrudates with 20%, 32%, and 35% amylose. An additional finding was that although the amylopectin promoted the expansion of the gelatinized starch matrix, it failed to strengthen and sustain the walls of the extrudate bubbles during expansion.     

Determination of amylopectin structure and physicochemical properties in rice endosperm starch of mutant lines derived from Malaysian rice cultivar MR219

BACKGROUND: Characterization of starch properties and functionality can apply breeding program selection for desirable traits such as eating, cooking and processing qualities to meet consumer preference. Low amylose content is generally preferred in Malaysia because of cohesive, tender and glossy cooked rice. Rice high in short‐chain amylopectin has a lower transition temperature of starch gelatinization. In the continuing search for improved starch quality in rice cultivars a study was carried out with new mutant lines MR219‐4 and MR219‐9, derived from MR219. RESULTS: MR219 and its mutant lines contain L‐type amylopectin, being high in amylopectin of intermediate chain length with degree of polymerization 12‐21. The apparent amylose content (AAC) in MR219‐4 had lower AAC value (19.2%) compared to other lines. A strongly negative correlation was found between chain‐length fraction of amylopectin and transition temperatures—onset temperature peak temperature, and conclusion temperature (0.992, 0.958, 0.950; P < 0.01)—with fraction b1 (fb1), respectively. CONCLUSION: The Malaysian lines studied contain L‐type amylopectin and offer a better understanding of grain quality improvement in terms of starch properties and functionality. This information will be directly applicable to select for desirable traits in future breeding programs. Copyright © 2012 Society of Chemical Industry     

Localisation of Amylose and Amylopectin in Starch Granules Using Enzyme-Gold Labelling

The independent localisation of amylose and amylopectin in a range of dry and hydrated native starch granules with varying amylose content (0—70 %) has been indirectly visualised using enzyme-gold cytochemical markers. Increasing amylose content was clearly demonstrated to result in distinct changes in granule architecture. In the absence of amylose (waxy maize starch) a framework of closely packed concentric layers of amylopectin exists in the granules. Low amylose content (potato starch) results in alternating layers of densely packed amylopectin and amylose molecules. High amylose content (amylomaize starch) granules were shown to possess an amylopectin centre surrounded by an amylose periphery encapsulated by an amylopectin surface. Elongated granules without the amylopectin centre were also observed in high amylose starches suggesting a relationship between amylopectin, amylose and granule shape. A model of starch granule architecture is proposed where increased compartmentalisation of amylose and amylopectin is observed in granules containing increasing levels of amylose.     

糯米对脾虚证模型大鼠的影响

通过给大鼠灌服大黄水煎液的方法制作脾虚证模型,然后比较糯米粉和糯米支链淀粉对脾虚证大鼠的一般生活状态、体质量及胃泌素含量的影响。结果发现与正常对照组相比,脾虚模型组大鼠生活状态较差,体质量、胃泌素含量明显下降(p<0.01);与脾虚模型组相比,糯米粉组和糯米支链淀粉组的大鼠生活状态明显改善,体质量增大,胃泌素含量明显升高(p<0.01),糯米支链淀粉的改善效果优于糯米粉。提示糯米粉具有改善脾虚症状的作用,这种作用与糯米中的支链淀粉有较大的相关性。      

The Effect of Environmental Temperature on Distribution of Unit Chains of Rice Amylopectin

The effects of environmental temperature during the early development of seeds on the structural characteristics of the endosperm starch were investigated using near‐isogenic lines of rice plants (Taichung 65, waxy), grown under temperature controlled conditions. High performance gel permeation chromatography (HPLC) demonstrated that Pseudomonas isoamylase‐debranched amylopectins of rice plants grown at lower temperature (25°C) contain increased amounts of short chains and decreased amounts of long chains as compared with amylopectins obtained from rice plants grown at higher temperature (30°C). By high performance anion exchange chromatography with pulsed amperometric detection (HPAEC‐PAD) of isoamylase‐debranched amylopectins it was detected that the amount of unit chains with degree of polymerization (DP) 6 and 11‐13, in the amylopectin of rice plants grown at lower temperature (25°C) had significantly increased and the amount of unit chains with DP 8, 22‐24 and 29 had significantly decreased, as compared with the amylopectin of rice plants grown at higher temperature (30°C). It was confirmed that the environmental temperature between 5 and 10 d after pollination strongly influenced the structure characteristics of the endosperm starch of rice plants.     

Morphological,thermal and rheological properties of starches separated from rice cultivars grown in India

Morphological, thermal and rheological properties of starches separated from five rice cultivars (PR-106, PR-114, IR-8, PR-103 and PR-113), varying in amylose content, were studied. Amylose contents of starches separated from PR-103, IR-8, PR-106, PR-114 and PR-113 were 7.83, 15.62, 16.05, 16.13 and 18.86%, respectively. The granular size, measured using a Scanning Electron Microscope, varied from 2.4 to 5.4 μm in all rice starches. PR-103 starch, with lowest average granular size, amylose content and solubility, had the highest swelling power, while PR-113 starch, with the highest average granular size and amylose content had the lowest swelling power. PR-103 starch showed highest transition temperatures, enthalpies of gelatinization, peak height index, range and enthalpies of retrogradation. The retrogradation (%) was observed to be highest in PR-113 starch and lowest in PR-103 starch. The changes in rheological parameters of rice starches during heating were measured using a Dynamic rheometer. PR-113 rice starch showed the highest G′, G″ and breakdown in G′ values, whereas PR-103 starch showed the lowest values for these parameters. Turbidity value of gelatinized pastes from all rice starches progressively increased up to the 3rd day during refrigerated storage, PR-103 starch paste showed the lowest turbidity value and PR-113 starch showed the highest value. The syneresis (%) of starch pastes, from different rice cultivars during storage at 4 °C, was also measured. The syneresis of starch pastes from all rice cultivars, except PR-103, increased with storage. PR-103 starch paste showed negligible syneresis during storage.     

The importance of amylose and amylopectin fine structures for starch digestibility in cooked rice grains

Statistically and causally meaningful relationships are established between starch molecular structures (obtained by size-exclusion chromatography, proton NMR and multiple-angle laser light scattering) and digestibility of cooked rice grains (measured by in vitro digestion). Significant correlations are observed between starch digestion rate and molecular structural characteristics, including fine structures of the distributions of branch (chain) lengths in both amylose and amylopectin. The in vitro digestion rate tends to increase with longer amylose branches and smaller ratios of long amylopectin and long amylose branches to short amylopectin branches, although the statistical analyses show that further data are needed to establish this unambiguously. These new relationships between fine starch structural features and digestibility of cooked rice grains are mechanistically reasonable, but suggestive rather than statistically definitive.     

Structure and Properties of Starch in Rice Double Mutants Lacking Starch Synthase (SS) IIa and Starch Branching Enzyme (BE) IIb

Starch biosynthetic enzymes form multi-protein complexes consisting of starch synthase (SS) I, SSIIa, and starch branching enzyme (BE) IIb, which synthesize amylopectin clusters. This study analyzed the starch properties in two double mutant rice lines lacking SSIIa and BEIIb, one of which expressed an inactive BEIIb protein. The ss2a be2b lines showed similar or greater seed weight than the be2b lines, and plant growth was not affected. The ss2a line showed increased short amylopectin chains resulting in a lower gelatinization temperature. Starch granule morphology and A-type crystallinity were similar between the ss2a line and the wild type, except for a mild chalky seed phenotype in the ss2a line. However, the starch phenotype of the ss2a be2b lines, which was similar to that of be2b but not ss2a, was characterized by increased long amylopectin chains, abnormal starch granules, and B-type crystallinity. The similarity in phenotype between the ss2a be2b and be2b lines may be attributed to the inability of the be2b mutants to generate short amylopectin branches, which serve as primers for SSIIa. Therefore, the presence or absence of SSIIa hardly affected the amylopectin structure under the be2b background. The amylose content was significantly higher in the ss2a be2b lines than in the be2b lines. Starch crystallinity was greater in ss2a be2b lines than in be2b lines, despite the fact that starch crystallinity is generally negatively correlated with amylose content. This suggests that the formation of a double helix between long amylopectin chains and amylose affects starch crystallinity in the ss2a be2b mutants.     

In-depth analysis of the endogenous reference genes used in the quantitative PCR detection systems for rice

To standardize the rice-specific PCR detection methods, five previously reported rice (Oryza sativa) taxon-specific genes were compared and evaluated. The investigated genes included the rice root-specific gene (gos9), the ppi phosphofructokinase gene (ppi-PPF), the phospholipase D gene (PLD), the starch branching enzyme gene (RBE4) and the sucrose phosphate synthase gene (SPS). Sequencing analyses revealed that among the tested rice cultivars, single-nucleotide polymorphisms (SNPs) existed in the gos9, PLD, ppi-PPF and SPS amplicons, though no statistically significant effect on their Ct values was found. The ppi-PPF and PLD systems were found to produce amplicons in non-rice species, such as sugarcane and broomcorn. The quantitative real-time PCR results revealed that this cross-reaction led to an underestimate of the GM (genetically modified) rice content. With the exception of the aforementioned shortcomings, these five endogenous reference genes all have acceptable amplification efficiencies, which ranged from 98 to 108?%, and high sensitivity within the limit of detection (LOD) values, which ranged from 5 to 10 copies of the haploid genome. In estimating the GM content in blinded rice samples, these five systems produce relatively accurate quantitative results with deviations less than 15?%, but the RBE4 system produced the most accurate quantitative results. Therefore, we have determined that the RBE4 gene is the most suitable rice reference gene, and the gos9 and SPS genes can also be used as rice reference genes because they have good commutability with the RBE4 gene. Care should be taken when interpreting results based on the PLD and ppi-PPF genes owing to their cross-reaction with other species.     

Comparison of Starch Physicochemical Properties from Medium‐Grain Rice Cultivars Grown in California and Arkansas

The starch molecular structure and physicochemical properties of two medium‐grain rice cultivars from Arkansas (Bengal, Medark) and from California (M202, M204) were determined and compared when they were grown in their respective locations and grown together in Arkansas to better understand the impacts of genetics and environment on starch characteristics. Both M202 and M204 contained significantly higher amylose contents (13.2‐15.3%) compared with the Arkansas cultivars (11.6‐12.4%). Starch from the Arkansas rice cultivars exhibited higher pasting and gelatinization temperatures and higher enthalpy values. Rice amylopectin from the California cultivars consisted of a smaller proportion of intermediate chains (DP 13‐24), and had a lower molecular weight and a smaller radius of gyration. When the four cultivars were grown together in Arkansas, the ranges for amylose content narrowed (10.6‐12.4%), their differences in thermal and pasting properties became smaller, and the molecular characteristics of amylose and amylopectin changed for all four cultivars. This study demonstrated that genetics, location, and crop year all contributed to variations in rice starch fine structure and physicochemical properties.     

糙米复配粉中淀粉直/支比对挤压速食粥品质与结构的影响

糙米是一种营养丰富的全谷物食品,但较差的适口性、蒸煮性和消化性制约着其成为主流餐桌食品。为改善其食用品质,本文通过分析不同淀粉直/支比(0.176、0.196、0.216、0.236、0.256、0.276)糙米复配粉的原料特性以及其对挤压速食粥糊化特性、复水品质、质构特性与结构的影响,探究制作糙米挤压速食粥原料复配粉最优的淀粉直/支比。结果表明:随着淀粉直/支比的增加,糙米复配粉的糊化温度和峰值温度增加,挤压速食粥米粒横截面的光滑度及紧密度增加。当淀粉直/支比为0.256时,挤压速食粥米粒横截面的结构最均一、最紧密,复水时间达到12.6 min,米汤固形物损失率和径向膨胀率最小(0.153 g和2.282%),此时的硬度、胶着度和耐咀性明显高于淀粉直/支比0.276,达到3323.85 g、1242.76 g·s和744.73 g。可见,糙米复配粉中淀粉直/支比0.256为制作挤压速食粥的较佳比例。     

Effect of Environmental Temperature During Development of Rice Plants on Some Properties of Endosperm Starch

Starch granules were isolated from polished grains of rice plants (Oriza sativa L.) grown under controlled temperature conditions. Distribution of isoamylase-debranched starch materials and unit chain-length of amylopectin were examined by gel-filtration on columns. Higher environment temperature decreases amylose concentration in endosperm starches of Japanese rice cultivars. The amylose concentration in the endosperm starch is determined by the environment temperature 5 – 15 days after heading. Interestingly, the higher environment temperature seemed to increase the amount of long B chains and decrease that of short chains of amylopectin. The effect of environment temperature on pasting characteristics of starch by DSC, starch-granule digestibility by amylases and X-ray diffractograms of starch was also investigated.