Effects of inhibition of starch branching enzymes on starch ordered structure and component accumulation in developing kernels of rice

Three rice varieties, Te-qing (TQ), Wu-xiang 9915 (WX) and Guang-ling-xiang-nuo (GLXN) with different amylose contents, and their derived lines with inhibition of starch branching enzyme I and IIb (SBEI/IIb) (GLXN-, WX- and TQ-SBEI/IIb^–) were used to investigate and compare the starch ordered structure and component accumulation in developing kernels. The starches in developing kernels of TQ, WX and GLXN all showed a typical A-type crystallinity and had similar short-range ordered structure, but their relative crystallinities gradually decreased in TQ and WX and had no significant change in GLXN with kernel development. For wild-type rices, starch components gradually increased in developing kernels. With inhibition of SBEI/IIb, the accumulation of amylose and amylopectin long branch-chains in kernels was slightly affected, but amylopectin short branch-chains seriously decreased, resulting in a significantly lower dry weight and starch content in kernel. The B-type crystallinity in developing starch gradually accumulated with kernel development of TQ-, Wx- and GLXN-SBEI/IIb^– lines. The inhibition of SBEI/IIb decreased the relative crystallinity but increased the short-range ordered structure of starches. The effects of inhibition of SBEI/IIb on starch ordered structure and component accumulation mainly appeared after 10 days after flowering, and were the most conspicuous for indica rice TQ. Though the inhibition of SBEI/IIb influenced the ordered structure and component accumulation of starch, the starch content and components in developing kernels all showed significantly positive correlations with kernel dry weight.     

The effects of puroindoline b on the ultrastructure of endosperm cells and physicochemical properties of transgenic rice plant

Endosperm texture is an important factor governing the end-product quality of cereals. The texture of wheat (Triticum aestivum L.) endosperm is controlled by puroindoline a and b genes which are both absent in rice (Oryza sativa L.). It has been reported that the endosperm texture of rice can be modified by puroindoline genes. The mechanism, however, by which puroindolines affect the ultrastructure of rice endosperm cells remains to be investigated. In this study, we observed the ultrastructure of endosperm cells and the morphology of isolated starch granules of the transgenic rice expressing the puroindoline b gene. SEM and TEM observations indicated that compound starch granules were embedded within the matrix material in non-transgenic rice, Nipponbare, whereas they were surrounded by spaces in the transgenic rice. The morphology and size of each starch granule were not different between non-transgenic and the transgenic rice. However, the transgenic rice flour showed smaller particle size, higher starch damage, and lower viscosity during gelatinization than that of non-transgenic rice. These results confirm that puroindoline b reduces the grain hardness in rice. Moreover, the results also suggest that puroindoline b functions at the surface of compound starch granules, and not on polygonal starch granules in rice endosperm.     

Hardness Distribution and Endosperm Structure on Polishing Characteristics of Brewer’s Rice Kernels

Abstract

This study was designed to determine the effects of the hardness distribution and the endosperm structure on the polishing characteristics of brewer’s rice kernels. We used four brewer’s rice cultivars, Kairyo-omachi, Hattan-nishiki No. 1, Senbon-nishiki and Yamada-nishiki. The broken kernel ratios in Kairyo-omachi and Hattan-nishiki No. 1 were significantly higher than those in Senbon-nishiki and Yamada-nishiki. Vickers hardness (VH) values in white-core tissues in kernels differed among varieties, which were significantly lower in Kairyo-omachi and Hattan-nishiki No. 1. However, no varietal differences were observed in VH values in the peripheral translucent tissues surrounding the white-core tissues. The tissues along the dorsoventral axis were softer than those along the longitudinal axis of the kernels. The tissues on the ventral side were softer than those on the dorsal side. Scanning electron microscopy (SEM) observations revealed the presence of closely arranged compound starch granules and few varietal differences in the peripheral translucent tissues surrounding the white-core tissues. However, as compared with Yamada-nishiki and Senbon-nishiki, in Hattan-nishiki No. 1 and Kairyo-omachi, the starch granules were loosely packed and the airspaces between the starch granules were more numerous in the white-core tissues. A higher number of airspaces and less starch were present in the endosperm cells along the dorsoventral axis when compared with along the longitudinal axis and on the ventral side than on the dorsal side. The present study showed that polishing characteristics are closely related with the endosperm structure, which is characterized as the density of starch granules.     

The Endosperm Morphology of Rice and its Wild Relatives as Observed by Scanning Electron Microscopy

While cultivated rice, Oryza sativa, is arguably the world’s most important cereal crop, there is little comparative morphological information available for the grain of rice wild relatives. In this study, the endosperm of 16 rice wild relatives were compared to O. sativa subspecies indica and O. sativa subspecies japonica using scanning electron microscopy. Although the aleurone, starch granules, protein bodies and endosperm cell shapes of the cultivated and non-cultivated species were similar, several differences were observed. The starch granules of some wild species had internal channels that have not been reported in cultivated rice. Oryza longiglumis, Microlaena stipoides and Potamophila parviflora, had an aleurone that was only one-cell thick in contrast to the multiple cell layers observed in the aleurone of the remaining Oryza species. The similarity of the endosperm morphology of undomesticated species with cultivated rice suggests that some wild species may have similar functional properties. Obtaining a better understanding of the wild rice species grain ultrastructure will assist in identifying potential opportunities for development of these wild species as new cultivated crops or for their inclusion in plant improvement programmes.     

Effects of added water and retrogradation on starch digestibility of cooked rice flours with different amylose content

Flours derived from rice varieties with different amylose content possess distinct physicochemical and molecular properties. The aim of this study was to determine optimal processing conditions for preparing rice flour-based foods with reduced starch digestibility. To do so, we evaluated the in vitro starch digestibility of rice flours with five varieties. Reducing the amount of water (from 10-fold to 4-fold of rice flour) used for cooking rice flour lowered its starch digestibility, and the magnitude of the decrease was positively correlated with amylose content. When retrogradation of cooked rice flour proceeded for 7 days, the digestibility of high-amylose rice flours declined rapidly in the first 3 days, whereas the digestibility of low-amylose rice flours declined continuously. Our analysis also demonstrated that the chain length distribution of starch molecules and the final and setback viscosity pasting properties were the most important parameters affecting the digestibility of rice flours. Based on our results, it appears possible to reduce rice starch digestibility by establishing optimum processing conditions for different varieties. We suggest a 7-fold addition of water and retrogradation for 1 day for high-amylose rice varieties and a 4-fold addition of water with 3 days of retrogradation for low-amylose rice.     

Investigation of the high-amylose maize starch gelatinization behaviours in glycerol-water systems

The effect of glycerol on gelatinization behaviours of high-amylose maize starch was evaluated by confocal laser scanning microscopy (CLSM), scanning electronic microscope (SEM), differential scanning calorimetry (DSC), texture analyzer (TPA) and rheometer. Gelatinization of the high-amylose maize starches with glycerol content of 10% (w/w) began at 95.4 °C (T_o), peaked at 110.3 °C (T_p), and completed at 118.9 °C (T_c). The birefringence began to disappear at around 100 °C and finished at 120 °C which corresponded well to the onset and conclusion temperatures obtained by DSC. The high-amylose maize starch granules maintained original morphological structure at 100 °C and swelled to a great degree at 110 °C. The high-amylose maize starch paste formed at 100 °C showed the lowest hardness (39.92 g), while at 120 and 130 °C, showed the highest hardness (610.89 g and 635.43 g, respectively). It should be noted that in going from 100 °C to 110 °C there is a significant increase in the viscosity of the slurry solution. The identical apparent viscosity was observed when the shear rate exceed 100 s⁻¹, resulting from the high-amylose maize starch granules were completely gelatinized at 120 °C, which was consistent with DSC analysis.     

Mapping QTLs for milling yield and grain characteristics in a tropical japonica long grain cross

Percent milling yield is an economically important trait of commercial rice because it largely determines the price that farmers receive for their crop. Analyzing 22 trait variables including milling yield, grain dimensions, chemistry and appearance, we identified 43 quantitative trait loci (QTLs) in a long grain japonica by long grain japonica cross. We report one QTL explaining 20% of the variation in brown rice recovery; two QTLs explaining 14% and 13% of the variation in milled rice recovery; and one QTL explaining 14% of the variation in head rice (HR) recovery. QTLs for the proportion of pre-broken brown rice kernels, seed density, amylose content, and kernel whiteness and chalkiness were found in the same region as the HR QTL. QTLs explaining up to 54% of the variation in grain shape measurements were identified and mapped to areas independent from those identified for milling yield. Analyses of grain appearance traits identified two QTLs for chalk in brown rice and one in head rice, and a QTL explaining up to 33% of the variance in green kernel area. Our results confirm previous findings on the multigenically complex nature of milling yield.     

玉米不同部位子粒灌浆特性与粒重的关系研究

以郑单958(ZD958)和登海661(DH661)为试验材料,研究在4.5万、7.5万、10.5万株/hm2种植密度下玉米上部及中下部子粒灌浆特性和粒重的关系。结果表明,穗粒数和千粒重是决定子粒最终产量的重要因素,两者均随着密度的增加而显著降低。同一密度下,穗位层光照自开花当天(0 d)先降低后升高,总体呈"高-低-高"的变化趋势,底层光照则持续升高。上部子粒的起始生长势、灌浆速率、胚乳细胞数及淀粉含量均低于中下部子粒。相关及通径分析表明,影响粒重的主要因素是灌浆速率、胚乳细胞数目和淀粉含量,其相关系数分别为0.959 66**、0.981 91**和0.877 76*。光照(包括穗位层和底层)主要通过灌浆速率和淀粉含量来间接影响粒重。     

Starch granule morphology in oat endosperm

Mature and developing oat (Avena sativa) grains were sectioned and image analysis methods used to estimate the starch granule-size distribution and morphology in endosperm cells. This showed that oat endosperm cells contain two types of starch granule: compound starch granules such as those seen in rice endosperm and in most other grasses; and simple granules similar to the B-type starch granules seen in the endosperm of Triticeae species such as wheat (Triticum aestivum). The simple granules in oats are similar in size and relative abundance to B-type granules in Triticeae suggesting that they may share a common evolutionary origin. However, there is a fundamental difference between oats and Triticeae in the timing of granule initiation during grain development. In Triticeae, the B-type granules initiate several days after the A-type granules whereas in oats, both the simple and compound granule types initiate at the same time, in early grain development.     

Endosperm Structure of White-Belly and White-Core Rice Grains Shown by Scanning Electron Microscopy

Abstract:

White-belly and white-core are the major two types of grain chalkiness in japonica rice. This study aims to compare the morphological features of white-belly and white-core using a scanning electron microcope (SEM). A japonica rice cultivar Wuyujing3 and its mutants were used as materials. Nearly 1000 SEM images were observed, and 12 representative photos were selected. SEM images showed contrasting differences between white-belly and white-core in endosperm microstructure including the shape of endosperm cell, the size distribution of starch granules, and the amount of protein bodies. White-belly and white-core also varied markedly in morphological features of the cracked compound starch granules. Our findings should help to advance our understanding of the multi-faceted nature of grain chalkiness from the perspective of starch and protein accumulation, and should be of value for future work on rice grain chalkiness.     

Difference in the Physical Properties of White-Core and Non-White-Core Kernels of the Rice Varieties for Sake Brewing is Unrelated to Starch Properties

Abstract

This study was designed to determine whether or not the difference in the physical properties between white-core and non-white-core kernels of the rice varieties for sake brewing is associated with their starch properties. We used two rice cultivars for sake brewing, Senbon-nishiki and Yamada-nishiki, from three different plots in Hiroshima prefecture. Hardness values of kernels were significantly higher in non-white-core than in white-core kernels in both varieties. Vickers hardness (VH) values were lowest at the center of the kernel in both types of kernels. VH values of white-core tissues of white-core kernels were significantly lower than those of corresponding tissues of non-white-core kernels. No significant differences were observed between the two types of kernels in VH values of the surrounding translucent tissues and in the starch properties (amylose content, pasting properties analyzed using a rapid viscoanalyzer and gelatinization properties analyzed using a differential scanning calorimetry). These results suggest that the difference in physical properties between the two types of kernels of the rice varieties for sake brewing are associated with the difference in structure of endosperm cells and not in starch properties.     

Field emission scanning electron and atomic force microscopy,and Raman and X-ray photoelectron spectroscopy characterization of near-isogenic soft and hard wheat kernels and corresponding flours

Secondary field emission scanning electron microscopy (FE SEM), atomic force microscopy (AFM), Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) were used to investigate native near-isogenic soft and hard wheat kernels and their roller milled flours. FE SEM images of flat-polished interior endosperm indicated distinct differences between soft and hard wheats with less internal continuity in the soft wheat, whereas individual starch granules were much less evident in the hard kernel due to a more continuous matrix. AFM images revealed two different microstructures. The interior of the hard kernel had a granular texture with distinct individual spheroid features of 10–50 nm while the images obtained for the soft kernel revealed less distinct small grains and more larger features, possibly micro-structural features of starch granules. Raman spectra resolved identical distinct frequencies for both kernel types with slightly different intensities between types. Finally, the chemical surface compositions of flour for these two types of kernels obtained by XPS provided subtle insight into the differences between soft and hard wheat kernels. These combined advanced microscopic and spectroscopic analyses provide additional insight into the differences between the soft and hard wheat kernels.     

Relative nutritional value of some rices for growth ofTribolium castaneum larvae

Growth of red flour beetleTribolium castaneum (Herbst) larvae with brown or milled rice as the carbohydrate source was faster on diets containing milled rice than on those with brown rice. Larval growth was negatively correlated with amylose content of both brown and milled rice. Among high-amylose (>25%) milled rices, heavier larvae were obtained with rices of low gelatinization temperature (alkali-spreading values 6–7) than with those with higher gelatinization temperature (alkali-spreading values < 6). The differences in larval growth reflected relative digestibility of raw rice starch granules.     

Influence of environmental temperature during grain filling period on granule size distribution of rice starch and its relation to gelatinization properties

High temperature (HT) is the major environmental factor affecting grain starch properties of cooking rice cultivars. However, little information has been available on the effect of environmental temperature on the starch granule size distribution of rice grains. In this paper, five indica rice genotypes, including the wild type (9311) and its four mutants differing in amylose content (AC), were used to investigate the effect of environmental temperature on the starch granule size distribution, as well as its relation to AC and gelatinization properties of rice starch. Two temperature treatments (HT and NT) at filling stage were imposed to rice plants under the controlled temperature chambers. The result showed that HT increased the average diameter of starch granules and enhanced the proportion of large starch granules (LSG, D > 2.6 μm) by number, volume and surface area, respectively. However, influence of HT on GT and starch granule size distribution was relatively independent of their alteration in AC level for different rice genotypes. Therefore, HT-induced increase in the average diameter of starch granules and LSG percentage was strongly responsible for the higher starch gelatinization temperature and inferior cooked palatability of HT-ripening rice grains, which be not inherently associated with their varying AC level.     

Starch granules size distribution in superior and inferior grains of wheat is related to enzyme activities and their gene expressions during grain filling

Mature wheat endosperm contains A-, B-, C-type starch granules, and each class has unique physiochemical properties which determine the quality of starch. The dynamics of the starch granule size distribution, activities of starch synthases and expression of starch synthase encoding genes were studied in superior and inferior grains during grain filling. Compared with inferior grains, superior grains showed higher grain weight, contents of starch, amylose and amylopectin. The formation of A-, B-, C-type starch granules initiated at 4, 8, 20 DAF, respectively, and was well consistent with the temporally change patterns of starch synthase activities and relative gene expression levels. For instance, activities of soluble and granule-bound starch synthases (designated SSS and GBSS) peaked at 20 and 24 DAF. Genes encoding isoforms of starch synthases expressed at different grain filling periods. In addition, SS I was generally expressed over the grain filling stage; the SS II and SS III were expressed over the early and mid grain filling stage, and the GBSS I was expressed during the mid to late grain filling stage. In addition, the time-course changes in activities of starch synthases and expression of starch synthase encoding genes explained well the dynamics of the starch granule size distribution.     

Biochemical characterization of QTLs associated with endosperm modification in quality protein maize

Genetic analysis using quality protein maize (QPM) recombinant inbred lines derived from K0326Y QPM and W64Ao2 identified three quantitative trait loci (QTL) in bins 1.06, 7.02 and 9.03 associated with opaque2 endosperm modification. We evaluated the effects of these QTLs on protein accumulation and starch physicochemical properties. The QTL in bin 1.06 is close to α-zein genes, and vitreous individuals with this QTL had increased accumulation of 19-kDa α-zein, 27-kDa γ-zein and legumin-1. The QTL in bin 7.02 corresponds to the γ-zein locus, and greater accumulation of this protein was found in vitreous individuals. The QTL in bin 9.03 is close to starch biosynthetic genes; greater accumulation of granule-bound starch synthase and amylose was observed in vitreous kernel samples with this locus and that in bin 1.06, as well as less gelatinization enthalpy and crystallinity. Vitreous kernels contained angular-shaped/compact starch granules and more short-intermediate length chains of amylopectin. These results support that endosperm modification in QPM is associated with increased accumulation of γ-zein and other storage proteins, but also show that synthesis of less crystalline starch with more amorphous regions at the periphery of granules, which favor their packing and association with endosperm proteins, may also be an important factor.     

Effects of heat,drought and their combined effects on morphological structure and physicochemical properties of rice (Oryza sativa L.) starch

High temperature (HT) and drought stress (WS) severely affect rice quality by altering the starch structure in rice. The morphological and physicochemical properties of starches isolated from two rice varieties grown under three stress treatment (HT, WS and WS + HT) during the grain filling stage were investigated. The results showed that WS increased amylose content (AC%) and the proportion of large starch granules (LSG) and made the surface of the starch granule smooth and flat. As a result, a lower relative crystallinity, surface order, swelling power, setback viscosity and gelatinization enthalpy were caused. HT decreased AC% and milled rice rate, but increased chalky rice rate, the number of LSG and the large air space and pits on the surface of the starch granules. As a result, a higher relative crystallinity, surface order, swelling power, setback viscosity and gelatinization enthalpy were caused. Similar results were observed under the treatment of WS + HT, indicating that there is a mild antagonistic effect on rice starch when the HT and WS occur simultaneously.     

Relationships of Ethylene Evolution Rate and 1-Aminocylopropane-1-Carboxylic Acid Concentration in Grains during Filling Period with Appearance Quality of Rice

To elucidate the relationship between ethylene evolution from the grains and the appearance quality of rice,ten different rice genotypes were used to determine the ethylene evolution rate,1-aminocylopropane-1-carboxylic acid(ACC)concentration in grains during grain filling and the appearance quality of rice,and the effects of chemical regulators on concentrations of ethylene and ACC in the grains during grain filling were also investigated to verify the roles of ethylene in the rice quality formation.The ethylene evolution rates and ACC concentrations in grains during the mid and late grain filling stages were very significantly and positively correlated with chalky kernel percentage and chalkiness.The cultivars with a low ACC concentration in grains exhibited a close amyloplast arrangement and small space between starch granules,whereas those with a high ACC concentration in grains showed a loose arrangement and wide space between the granules.Application of 1μmol/L ACC to panicles at mid and late grain filling stages significantly loosened amyloplast arrangement and increased chalky kernel percentage,chalky area and chalkiness,and the results were reversed when 1μmol/L amino-ethoxyvinylglycine,an inhibitor of ACC synthesis enzyme,was applied to panicles.A practice of moderate dry-wet alternate irrigation reduced ethylene evolution and ACC concentration in grains and thereby reduced chalkiness. The results suggested that ethylene and ACC in grains play an important role in the endosperm structure and appearance quality of rice,and the appearance quality would be improved by reducing ethylene evolution and ACC in grains through either variety breeding and selection,or chemical regulations or cultivation techniques.     

Analyses of albumins, globulins and amphiphilic proteins by proteomic approach give new insights on waxy wheat starch metabolism

Starch is composed of two types of glucose polymers: amylose and amylopectin. The Waxy (Wx) locus controls amylose synthesis in the wheat kernel. Hexaploid wheat has three Wx loci located on chromosomes 7A (Wx-A1), 4A (Wx-B1), and 7D (Wx-D1). Eight near isogenic lines (NILs) of Triticum aestivum cv. Tremie with one, two or three Wx null alleles were used. The albumin–globulin fraction, and amphiphilic proteins were separated using 2-dimensional electrophoresis (2DE) allowing the changes in the waxy kernel to be identified. Albumin–globulin fraction showed overexpression of sucrose synthases in the waxy NILs compared to the normal form of Tremie and a decrease in many proteins related to stress and defence metabolism such as serpins. A subunit of ADP-glucose pyrophosphorylase (AGPase), which is known to play a major role in starch synthesis, was also shown to be down regulated in the waxy NILs. Amphiphilic proteins confirmed the observations made on the albumin–globulin fraction with a decrease in a stress-related protein. These different regulations linked to observations made on wheat kernel (thousand kernel weight (TKW), protein amount per grain, size and distribution of the starch granules) led to formulation of the hypothesis that waxy endosperm does not reach maturity of the wild-type endosperm.