Structures, gelatinization properties and enzyme hydrolyses of starches from transparent and floury grains of rices subjected to field natural extreme high temperature.

Three rice varieties underwent the field natural extreme high temperature (EHT) with daily average temperature over 30 °C from 21 to 89 days after sowing, and had transparent, chalky and floury grains. The structures, gelatinization properties and enzyme hydrolyses of starches from transparent and floury grains were investigated. Compared with control transparent grains, floury grains subjected to EHT markedly decreased the contents of amylose molecules, amylopectin A chains and amylopectin B1 chains and increased the contents of amylopectin B2 and B3+ chains and the average branch-chain length of amylopectin. Both transparent and floury grains had A-type starches, but floury grain starches exhibited higher relative crystallinity, gelatinization temperature, retrogradation and pasting viscosities than transparent grain starches. Floury grain starches had lower hydrolysis rates than transparent grain starches. Native starches were more resistant to digestion but gelatinized and retrograded starches were more prone to digestion in floury grains than in transparent grains.

Identification and analysis of nine new flo2 allelic mutants in rice.

FLO2 is involved in grain development and storage substance synthesis in rice, and therefore can regulate grain size and quality. In this study, we identified 4 new flo2 allelic mutants with nonsense and frameshift mutation in the exon of 6, 10, 11 and 21 and 5 new flo2 allelic mutants with alternative splicing and frameshift mutation at the splicing site of intron 13, 14, 16 and 17. Compared with wild-type rice, the outer endosperm of flo2 mutants was transparent, and the inner endosperm was floury. Different mutation sites and types of FLO2 significantly decreased kernel width, thickness and weight to some extent. The contents of storage protein, starch, amylose and amylopectin showed significant decrease at different levels among 9 flo2 mutants. The expressions of most storage protein synthesis genes and starch synthesis-related genes were significantly down-regulated, and exhibited different ranges of variation among different flo2 mutants. This study could add helpful information for the roles of flo2 alleles in rice quality regulation and provide abundant germplasm resources for rice quality breeding.

Physicochemical properties of starches from sweet potato root tubers grown in natural high and low temperature soils.

Three sweet potato varieties grew in natural high temperature (HT) and low temperature (LT) field soils. Their starch physicochemical properties were affected similarly by HT and LT soils. Compared with LT soil, HT soil induced the increases of granule size D[4,3] from 18.0-18.7 to 19.9-21.8 µm and amylopectin average branch-chain length from 21.9-23.1 to 24.1-24.7 DP. Starches from root tubers grown in HT and LT soils exhibited CA- and CC-type XRD pattern, respectively. Starches from root tubers grown in HT soil exhibited stronger lamellar peak intensities (366.8-432.0) and higher gelatinization peak temperature (72.0-76.8 °C) than those (176.2-260.5, 56.4-63.4 °C) in LT soil. Native starches from root tubers grown in LT soil were hydrolyzed more easily (hydrolysis rate coefficient 0.227-0.282 h-1) by amylase than those (0.120-0.163 h-1) in HT soil. The principal component analysis exhibited that starches from root tubers grown in HT and LT soils had significantly different physicochemical properties.

Effects of growth temperature on multi-scale structure of root tuber starch in sweet potato.

Sweet potato was planted at three soil and air temperatures (21, 25 and 28 °C) with the same humidity and light time/intensity. Root tuber starches were isolated, and their multi-scale structures were investigated to reveal the effects of growth temperature on starch properties. Growth temperature did not change the morphology and amylose content of starch, but markedly increased the size of starch from volume-weighted mean diameter 12.2 µm to 17.0 µm. Starch grown at high growth temperature exhibited less A branch-chains and lower branching degree of amylopectin and more B2 and B3+ branch-chains of amylopectin than at low growth temperature. With increasing growth temperature, starch changed from CC-type to CA-type, its relative crystallinity and lamellar peak intensity increased, and the thickness of crystalline and amorphous lamellae did not significantly change. Starch grown at high growth temperature exhibited significantly higher gelatinization temperature than at low growth temperature, but had similar gelatinization enthalpy.

Relationships between X-ray Diffraction Peaks, Molecular Components, and Heat Properties of C-Type Starches from Different Sweet Potato Varieties.

C-type starches with different proportions of A- and B-type crystallinities have different intensities and crystallinities of X-ray diffraction peaks. In this study, the intensities and crystallinities of X-ray diffraction peaks, molecular components and heat properties of C-type starches were investigated in seven sweet potato varieties, and their relationships were analyzed. The intensity and crystallinity of a diffraction peak at 5.6° were significantly positively correlated to the DP6-12 branch-chains of amylopectin and significantly negatively correlated to the true amylose content (TAC) determined by concanavalin A precipitation, gelatinization temperature, gelatinization enthalpy, water solubility at 95 °C, and pasting temperature. The intensity of diffraction peaks at 15° and 23° were significantly positively correlated to the gelatinization temperature and pasting temperature and significantly negatively correlated to the pasting peak viscosity. The significantly positive relationships were detected between the crystallinity of a diffraction peak at 15° and the DP13-24 branch-chains of amylopectin, gelatinization conclusion temperature and water solubility, between the crystallinity of diffraction peak at 17-18° and the TAC, gelatinization onset temperature, water solubility and pasting temperature, between the crystallinity of a diffraction peak at 23° and the gelatinization conclusion temperature and pasting peak time, and between the total crystallinity and the TAC, gelatinization conclusion temperature, water solubility and pasting temperature. The score plot of principle component analysis showed that the molecular components and heat property parameters could differentiate the C-type starches and agreed with their characteristics of X-ray diffraction peaks. This study provides some references for the utilizations of C-type starches.

Sizes, Components, Crystalline Structure, and Thermal Properties of Starches from Sweet Potato Varieties Originating from Different Countries.

Sweet potato is a root tuber crop and an important starch source. There are hundreds of sweet potato varieties planted widely in the world. Starches from varieties with different genotype types and originating from different countries have not been compared for their physicochemical properties. In the research, starches from 44 sweet potato varieties originating from 15 countries but planted in the same growing conditions were investigated for their physicochemical properties to reveal the similarities and differences in varieties. The results showed that the 44 starches had granule size (D[4,3]) from 8.01 to 15.30 µm. Starches had different iodine absorption properties with OD680 from 0.259 to 0.382 and OD620/550 from 1.142 to 1.237. The 44 starches had apparent amylose content from 19.2% to 29.2% and true amylose content from 14.2% to 20.2%. The starches exhibited A-, CA-, CC-, or CB-type X-ray diffraction patterns. The thermograms of 44 starches exhibited one-, two-, or three-peak curves, leading to a significantly different gelatinization temperature range from 13.1 to 29.2 °C. The significantly different starch properties divide the 44 sweet potato varieties into different groups due to their different genotype backgrounds. The research offers references for the utilization of sweet potato germplasm.

Effects of Variety and Growing Location on Physicochemical Properties of Starch from Sweet Potato Root Tuber.

Three sweet potato varieties with purple-, yellow-, and white-fleshed root tubers were planted in four growing locations. Starches were isolated from their root tubers, their physicochemical properties (size, iodine absorption, amylose content, crystalline structure, ordered degree, lamellar thickness, swelling power, water solubility, and pasting, thermal and digestion properties) were determined to investigate the effects of variety and growing location on starch properties in sweet potato. The results showed that granule size (D[4,3]) ranged from 12.1 to 18.2 µm, the iodine absorption parameters varied from 0.260 to 0.361 for OD620, from 0.243 to 0.326 for OD680 and from 1.128 to 1.252 for OD620/550, and amylose content varied from 16.4% to 21.2% among starches from three varieties and four growing locations. Starches exhibited C-type X-ray diffraction patterns, and had ordered degrees from 0.634 to 0.726 and lamellar thicknesses from 9.72 to 10.21 nm. Starches had significantly different swelling powers, water solubilities, pasting viscosities, and thermal properties. Native starches had rapidly digestible starch (RDS) from 2.2% to 10.9% and resistant starch (RS) from 58.2% to 89.1%, and gelatinized starches had RDS from 70.5% to 81.4% and RS from 10.8% to 23.3%. Two-way ANOVA analysis showed that starch physicochemical properties were affected significantly by variety, growing location, and their interaction in sweet potato.

Physicochemical properties of a new starch from ramie (Boehmeria nivea) root.

A new starch was isolated from ramie root, and its physicochemical properties were investigated. Ramie dry root contained 45.9% starch. Starch had truncated, ellipsoidal, and spherical granule shapes with size from 7 to 30 µm and D[4,3] about 14.1 µm. Starch contained 38.9% apparent amylose content and 22.4% true amylose content, exhibited B-type crystallinity, and had 26.6% relative crystallinity, 0.82 ordered degree, and 9.2 nm lamellar thickness. Starch had 71.8 °C gelatinization peak temperature and 15.6 J/g gelatinization enthalpy, and exhibited 31.4 g/g swelling power and 17.1% water solubility at 95 °C. Starch had peak, hot, breakdown, final, and setback viscosities at 3048, 2768, 279, 4165, and 1397 mPa s, respectively, and showed peak time at 4.36 min and pasting temperature at 75.0 °C. The native, gelatinized, and retrograded starches contained 15.1%, 94.0%, and 86.5% rapidly digestible starch and 83.3%, 4.0%, and 10.7% resistant starch, respectively. Compared with potato and rice starches, ramie starch was somewhat similar to potato starch but significantly different from rice starch in starch component, crystalline structure, and functional properties. Therefore, ramie starch exhibited the potential to be used as a thickening agent, resistant-digesting food additive, and alternative to potato starch in food and nonfood industries.

The defective effect of starch branching enzyme IIb from weak to strong induces the formation of biphasic starch granules in amylose-extender maize endosperm.

KEYMESSAGE: Biphasic starch granules in maize ae mutant underwent the weak to strong SBEIIb-defective effect during endosperm development, leading to no birefringence in their exterior due to extended long branch-chains of amylopectin. Biphasic starch granules are usually detected regionally in cereal endosperm lacking starch branching enzyme (SBE). However, their molecular structure, formation mechanism, and regional distribution are unclear. In this research, biphasic starch granules were observed in the inner region of crown endosperm of maize ae mutant, and had poorly oriented structure with comb-like profiles in their exterior. The inner endosperm (IE) rich in biphasic starch granules and outer endosperm (OE) without biphasic starch granules were investigated. The starch had lower amylose content and higher proportion of long branch-chains of amylopectin in IE than in OE, and the exterior of biphasic starch granules had less amylose and more long branch-chains of amylopectin than the interior. Compared with OE, the expression pattern of starch synthesis related enzymes changed significantly in IE. The granule-bound starch synthase I activity within biphasic starch granules decreased slightly. The IE experienced more severe hypoxic stress than OE, and the up-regulated anaerobic respiration pathway indicated an increase in carbon consumption. The starch in IE underwent the SBEIIb-defective effect from weak to strong due to the lack of sufficient carbon inflow, leading to the formation of biphasic starch granules and their regional distribution in endosperm. The results provided information for understanding the biphasic starch granules.

Starch Components, Starch Properties and Appearance Quality of Opaque Kernels from Rice Mutants.

Rice mutants with altered starch components and properties are important genetic resources in rice breeding programmes. In this study, 44 mutants with altered starch components were screened from 135 rice mutants with opaque kernels using a starch-iodine absorption spectrum method, and nine mutants from them were further selected for investigating their starch properties and kernel appearance quality. The results showed that the iodine absorption spectrum parameters, OD620, OD620/550, and λmax, could reflect the changes of starch components in rice mutants, and had significantly positive relationships with amylose content and negative relationships with the proportion of short branch-chains of amylopectin. The endosperm starches from nine mutants all showed A-type crystalline structure and similar short-range ordered structure, but had different relative crystallinities. The changes of starch components in mutants not only resulted in the different gelatinization properties of starch but also changed the appearance quality of brown rice kernels. This study provided abundant genetic plants for studying the molecular mechanism of starch synthesis and the quality regulation of rice kernels.

Effects of inhibiting starch branching enzymes on molecular and crystalline structures of starches from endosperm different regions in rice.

Mature endosperm was separated regionally into different parts in three rice cultivars, Te-qing (TQ), Wu-xiang 9915 (WX9915) and Guang-ling-xiang-nuo (GLXN), and their transgenic lines with inhibition of starch branching enzyme I and IIb (SBEI/IIb-). Within the three wild-type cultivars, starches from endosperm different regions showed similar molecular and crystalline structures. However, in rices with inhibition of SBEs, amylopectin short branch-chain content and branching degree gradually decreased, but amylopectin B3+ chain content and average chain length increased gradually from the interior to exterior of endosperm. The amylose content gradually increased from the interior to exterior of endosperm in TQ- and WX9915-SBEI/II- lines. From the interior to exterior of endosperm, starch changed gradually from CC- to CB-type in TQ-SBEI/II- line and from CA- to CC-type in GLXN-SBEI/II- line, and remained CA-type in WX9915-SBEI/II- line. These results provided some information for quality breeding and utilizations of rice with inhibition of SBE.

Cooking, morphological, mechanical and digestion properties of cooked rice with suppression of starch branching enzymes.

Kernel components and some physicochemical properties of cooked rice were investigated and compared between a popular japonica rice Wu-xiang 9915 (WX) and its transgenic line (WX-SBEI/IIb-) with suppression of starch branching enzyme I/IIb. The starch content, especially amylopectin content, was significantly lower in WX-SBEI/IIb- than in WX. Brown rice flour had markedly higher gelatinization temperature in WX-SBEI/IIb- than in WX. The cooked kernels of WX-SBEI/IIb- had significantly lower volume swelling, leached material amount and wet weight than those of WX during cooking. Starch granules in WX kernel could be gelatinized completely and gradually from the exterior to the interior of endosperm, leading to breakage of cooked kernels. However, aggregate, elongated and small starch granules in the exterior of WX-SBEI/IIb- endosperm could not be gelatinized completely and remained their morphologies during cooking, leading to a high resistance of kernels to cooking. Brown rice flour of WX-SBEI/IIb- had significantly lower pasting viscosities, storage modulus and loss modulus but higher loss angle tangent than that of WX. The cooked kernels of WX-SBEI/IIb- had considerably higher hardness, springiness and cohesiveness but lower adhesiveness than those of WX. The starch in cooked kernels was more resistant to digestion in WX-SBEI/IIb- than in WX.

Physicochemical properties of starches from vitreous and floury endosperms from the same maize kernels.

Starches were isolated from vitreous and floury endosperms from the same kernels, and their physicochemical properties were investigated to reveal their differences. Starch from vitreous endosperm had more surface indentations, larger granule size, and higher contents of protein, amylose and damaged starch than that from floury endosperm. Both starches from vitreous and floury endosperms exhibited A-type crystalline structure, but starch from vitreous endosperm had lower relative crystallinity and lamellar peak intensity than that from floury endosperm. Starch from floury endosperm had higher gelatinization temperature and enthalpy but lower gelatinization temperature range and percentage of retrogradation than that from vitreous endosperm. Native starch from floury endosperm was more resistant to enzyme hydrolysis than that from vitreous endosperm. Gelatinized and retrograded starches had similar digestion properties between starches from vitreous and floury endosperms. Principal component analysis indicated that starches from vitreous and floury endosperms had significant differences in their physicochemical properties.

Inhibition of starch branching enzymes in waxy rice increases the proportion of long branch-chains of amylopectin resulting in the comb-like profiles of starch granules.

Starches with comb-like profiles have been detected in some cereal endosperms with inhibiting expression of starch branching enzyme (SBE). Although amylose is considered to be an important factor in the formation of the comb-like profile, the details remain unclear. In this study, a transgenic rice line (GLXN-SBEI/IIb-) was derived from japonica waxy rice cultivar Guang-ling-xiang-nuo (GLXN) through antisense RNA inhibition of both SBEI and SBEIIb. The expression and activity of SBEI, SBEIIb and SBEIIa were declined. The GLXN-SBEI/IIb- endosperm contained large and small starch granules, and these starch granules had the comb-like profiles. The comb-like profiles of starches were detected in GLXN-SBEI/IIb- endosperm after 10 days after flowering with gradually increasing proportion of long branch-chains of amylopectin. The long branch-chains of amylopectin were responsible for forming the comb-like profiles at the outer region of starch granules. The gradually decreasing expression of SBEs influenced the synthesis of amylopectin during endosperm development, resulting in different structure between the inner and outer regions of starch granules from GLXN-SBEI/IIb- endosperm. The above results indicated that the long branch-chains of amylopectin, not amylose, led to the formation of comb-like profiles of starch granules in cereal crops with inhibiting expression of SBEs.

In situ Degradation and Characterization of Endosperm Starch in Waxy Rice with the Inhibition of Starch Branching Enzymes during Seedling Growth.

High-resistant starch cereal crops with the inhibition of the starch branching enzyme (SBE) have been widely studied. However, the effects of the inhibition of SBE on waxy cereal crops are unclear. A transgenic rice line (GTR) derived from a japonica waxy rice cultivar Guang-ling-xiang-nuo (GLXN) has been developed through antisense RNA inhibition of both SBEI and SBEIIb. In this study, GLXN and GTR were cultivated in the dark only in deionized H2O, and their shoot and root growth, starch in situ degradation, and starch property changes were investigated during seedling growth. Compared with GLXN, GTR showed a significantly slow seedling growth, which was not due to the embryo size and vitality. The slow degradation of starch in the seed restrained the seedling growth. GLXN starch was completely degraded gradually from the proximal to distal region of the embryo and from the outer to inner region in the endosperm, but GTR starch in the peripheral region of the endosperm was not completely degraded, and the starch residual was located in the outside of the compound starch though its degradation pattern was similar to GLXN. During seedling growth, GLXN starch had the same A-type crystallinity and a similar ordered structure, but the crystallinity changed from the CA-type to B-type and the ordered structure gradually increased in the GTR starch. The above results indicated that GTR had a heterogeneous starch distributed regionally in the endosperm. The starch in the peripheral region of the endosperm had a B-type crystallinity, which was located in the outside of the compound starch and significantly increased the resistance to in situ degradation, leading to the seedling slow growth.

Comparison of Physicochemical Properties of Starches from Flesh and Peel of Green Banana Fruit.

Green banana fruit is an important starch resource that consists of flesh and peel. The physicochemical properties of flesh starch have been widely studied; however, those of peel starch have hardly been studied, leading to the waste of peel. In this study, the physicochemical properties of the starches from the flesh and peel of green banana fruit were investigated and compared. The dry flesh and peel had 69.5% and 22.6% starch content, respectively. The starch had oval and irregular granules with eccentric hila. Their starches had similar bimodal size distribution; the volume-weighted mean diameter was approximate 17 µm, and the peel starch had a slightly smaller granule size than the flesh starch. The maximum absorption wavelength was higher in peel starch than in flesh starch. The apparent amylose content of flesh and peel starch was 21.3% and 25.7%, respectively. The flesh and peel starches both exhibited B-type crystalline structures and had similar relative crystallinity, short-range ordered degrees, and lamellar structures. The swelling power was similar between flesh and peel starches, but the water solubility was higher in peel starch than in flesh starch at 95 °C. The peel starch had a higher gelatinization temperature than flesh starch, but their gelatinization temperature range and enthalpy were similar. Both flesh and peel starches showed a diphasic hydrolysis dynamic, but peel starch had higher resistance to porcine pancreatic α-amylase hydrolysis than flesh starch. The contents of rapidly digestible starch, slowly digestible starch, and the resistant starch of flesh and peel were 1.7%, 4.3%, 94.1% and 1.4%, 3.4%, 95.2%, respectively, for native starch, and 73.0%, 5.1%, 21.9%, and 72.3%, 4.5%, 23.2%, respectively, for gelatinized starch.

A Novel Mutation of OsPPDKB, Encoding Pyruvate Orthophosphate Dikinase, Affects Metabolism and Structure of Starch in the Rice Endosperm.

Starch, as a main energy storage substance, plays an important role in plant growth and human life. Despite the fact that several enzymes and regulators involved in starch biosynthesis have been identified, the regulating mechanism of starch synthesis is still unclear. In this study, we isolated a rice floury endosperm mutant M14 from a mutant pool induced by 60Co. Both total starch content and amylose content in M14 seeds significantly decreased, and starch thermal and pasting properties changed. Compound starch granules were defected in the floury endosperm of M14 seeds. Map-based cloning and a complementation test showed that the floury endosperm phenotype was determined by a gene of OsPPDKB, which encodes pyruvate orthophosphate dikinase (PPDK, EC 2.7.9.1). Subcellular localization analysis demonstrated that PPDK was localized in chloroplast and cytoplasm, the chOsPPDKB highly expressed in leaf and leaf sheath, and the cyOsPPDKB constitutively expressed with a high expression in developing endosperm. Moreover, the expression of starch synthesis-related genes was also obviously altered in M14 developing endosperm. The above results indicated that PPDK played an important role in starch metabolism and structure in rice endosperm.

Comparison of structural and functional properties of starches from five fruit kernels.

Starch was isolated from the kernels of jackfruit, longan, loquat, litchi, and mango fruits, which contained approximately 56, 59, 71, 53, and 64% starch, respectively, indicating that these fruit kernels are good starch sources. The structural and functional properties of these isolated starches were investigated and compared. The starches had irregular, truncated, spherical, and elliptical shapes with central hila and exhibited different sizes, with mango starch being the largest and jackfruit and longan starches being the smallest. The five starches had similar amylose contents but exhibited significantly different crystalline properties including crystalline type, relative crystallinity, short-range ordered structure, and lamellar intensity. Among the five starches, the jackfruit and loquat starches had the highest and lowest gelatinization temperature and enthalpy, respectively, and the litchi and mango starches had the highest and lowest pasting viscosity, respectively. The longan and loquat starches were more susceptible to enzyme hydrolysis than the other starches.

Effects of surface proteins and lipids on molecular structure, thermal properties, and enzymatic hydrolysis of rice starch

Abstract Rice starches with different amylose contents were treated with sodium dodecyl sulfate (SDS) to deplete surface proteins and lipids, and the changes in molecular structure, thermal properties, and enzymatic hydrolysis were evaluated. SDS treatment did not significantly change the molecular weight distribution, crystalline structure, short-range ordered degree, and gelatinization properties of starch, but significantly altered the pasting properties and increased the swelling power of starch. The removal of surface proteins and lipids increased the enzymatic hydrolysis and in vitro digestion of starch. The influences of removing surface proteins and lipids from starch on swelling power, pasting properties, and enzymatic hydrolysis were different among the various starches because of the differences in molecular structures of different starch styles. The aforementioned results indicated that removing the surface proteins and lipids from starch did not change the molecular structure but had significant effects on some functional properties.

Comparison of Structural and Functional Properties of Starches from the Rhizome and Bulbil of Chinese Yam.

Chinese yam is an important edible starch plant and widely cultivated in China. Its rhizome and bulbil are starch storage tissues below and above ground, respectively. In this paper, starches were isolated from the rhizome and bulbil of Chinese yam, and their structural and functional properties were compared. Both starches had an oval shape with an eccentric hilum and a CA-type crystalline structure. Their short-range ordered structure and lamellar structure had no significant difference. However, the rhizome starch had a significantly bigger granule size and lower amylose content than the bulbil starch. The swelling power and water solubility were significantly lower in the rhizome starch than in the bulbil starch. The onset and peak gelatinization temperatures were significantly higher in the rhizome starch than in the bulbil starch. The rhizome starch had a significantly higher breakdown viscosity and a lower setback viscosity than the bulbil starch. The thermal stability was lower in the rhizome starch than in the bulbil starch. The rhizome starch had a significantly lower resistance to hydrolysis and in vitro digestion than the bulbil starch. The above results provide important information for the utilization of rhizome and bulbil starches of Chinese yam.