Formation and Developmental Characteristics of A- and B-Type Starch Granules in Wheat Endosperm

Wheat grain natural transverse sections of 12 periods were observed and analyzed using scanning electron micrographs technology and Bio-Quant system IV image analyzer in order to detect the developing process of A- and B-type starch granules. In addition, the chemical composition and starch granule-bound proteins （SGPs） of A- and B-type starch granules were tested and analyzed. The results showed that A-type starch granules in wheat began from 3 d post anthesis （DPA） till grain maturing and B-type starch granules occured after 15 DPA till grain maturing. Approximately 98.5% of chemical compositions in both A- and B-type starch granules were amylose and amylopectin, and more than half of which were amylopectin. The amylopectin contents, average chain length, and chain length distribution （degree of polymerization〉 40） of amylopectin in A-type starch granules were significant higher than that of B-type starch granules. SGP-145, SGP-140, and SGP-26 kD were associated with A-type starch formation in wheat grain.

Characterization of A-and B-type starch granules in Chinese wheat cultivars

Starch is the major component of wheat flour and serves as a multifunctional ingredient in food industry. The objective of the present study was to investigate starch granule size distribution of Chinese wheat cultivars, and to compare structure and functionality of starches in four leading cultivars Zhongmai 175, CA12092, Lunxuan 987, and Zhongyou 206. A wide variation in volume percentages of A- and B-type starch granules among genotypes was observed. Volume percentages of A- and B-type granules had ranges of 68.4–88.9% and 9.7–27.9% in the first cropping seasons, 74.1–90.1% and 7.2–25.3% in the second. Wheat cultivars with higher volume percentages of A- and B-type granules could serve as parents in breeding program for selecting high and low amylose wheat cultivars, respectively. In comparison with the B-type starch granules, the A-type granules starch showed difference in three aspects：（1） higher amount of ordered short-range structure and a lower relative crystallinity,（2） higher gelatinization onset（To） temperatures and enthalpies（ΔH）, and lower gelatinization conclusion temperatures（Tc）,（3） greater peak, though, and final viscosity, and lower breakdown viscosity and pasting temperature. It provides important information for breeders to develop potentially useful cultivars with particular functional properties of their starches suited to specific applications.

Hydrolysis of Raw Corn Starch Granules by Glucoamylase and Product Inhibition During the Hydrolysis

Raw corn starch granules were hydrolysized by glucoamylase in a chemostat. The hydro- lysis of three different-sized granules shows that smaller granules undergo more hydrolyzation than larger ones. After 78 h, 9700 of the granules was hydrolysized with diameter between 0.15 mm and 0.3 mm at 50 ℃. When corn starch concentration increased from 100 g/L to 250 g/L, the amount of reducing sugar produced was proportional to the initial substrate concentration and no substrate inhibition phenomenon appeared. In order to study the product inhibition exactly, the product from hydrolysis reaction itself was added into the hydrolysis system at the beginning of starch hydrolysis. Product inhibition with different quantities of product added were studied in the initial several hours, during which period enzyme inactivation could be neglected and product inhibition could be studied separately. The experiments indicate that product inhibition happens when the additional quantity exceeds 9.56 g/L.

Irrigation regimes modulate non-structural carbohydrate remobilization and improve grain filling in rice(Oryza sativa L.)by regulating starch metabolism

Recently developed ‘super’ rice cultivars with greater yield potentials often suffer from the problem of poor grain filling, especially in inferior spikelets. Here, we studied the activities of enzymes related to starch metabolism in rice stems and grains, and the microstructures related to carbohydrate accumulation and transportation to investigate the effects of different water regimes on grain filling. Two ‘super’ rice cultivars were grown under two irrigation regimes of well-watered(WW) and alternate wetting and moderate soil drying(AWMD). Compared with the WW treatment,the activities of ADP glucose pyrophosphorylase(AGPase), starch synthase(StSase) and starch branching enzyme(SBE), and the accumulation of non-structural carbohydrates(NSCs) in the stems before heading were significantly improved, and more starch granules were stored in the stems in the AWMD treatment. After heading, the activities of α-amylase, β-amylase, sucrose phosphate synthase(SPS) and sucrose synthase in the synthetic direction(SSs)were increased in the stems to promote the remobilization of NSCs for grain filling under AWMD. During grain filling, the enzymatic activities of sucrose synthase in the cleavage direction(SSc), AGPase, StSase and SBE in the inferior spikelets were increased, which promoted grain filling, especially for the inferior spikelets under AWMD.However, there were no significant differences in vascular microstructures. The grain yield and grain weight could be improved by 13.1 and 7.5%, respectively, by optimizing of the irrigation regime. We concluded that the low activities of key enzymes in carbon metabolism is the key limitation for the poor grain filling, as opposed to the vascular microstructures, and AWMD can increase the amount of NSC accumulation in the stems before heading, improve the utilization rate of NSCs after heading, and increase the grain filling, especially in the inferior spikelets, by altering the activities of key enzymes in carbon metabolism.

Rice Grains from Slightly Saline Field Exhibited Unchanged Starch Physicochemical Properties but Enhanced Nutritional Values

This study aims to investigate grain quality and nutritional values of rice(Pokkali,a salt-tolerant cultivar;RD73,a new cultivar improved from KDML105 introgressed with Saltol QTL from Pokkali,and KDML105,a moderately salt-susceptible cultivar)grown under non-saline(0.04–0.87 dS/m)and slightly saline(1.08–4.83 dS/m)field conditions.The results revealed that salinity caused significant reduction in grain size but significant increments in reducing sugar and total protein contents in the grains.Nevertheless,the amounts of starch in the grains of KDML105 and Pokkali rice genotypes were unaffected by the stress.The starch granule size distribution was also unaffected by salinity.Interestingly,only starch from Pokkali was significantly diminished in amylose content,from 19.18%to 16.99%.Accordingly,parameters relating to starch gelatinization,retrogradation,and pasting properties of KDML105 and RD73 were unaffected by salinity;only Pokkali showed a significant increase in percentage of retrogradation along with a significant reduction in gelatinization enthalpy.In the saline field,total phenolic content and antioxidant capacity in the grains of all rice cultivars tended to increase,particularly in Pokkali.On average,essential element contents in grains from the saline-treated plants showed a 33%,32%,32%,22%,20%,11%,and 10%increase in total P,N,K,Mg,Zn,Fe,and Ca content,respectively.Interestingly,total Fe content exhibited the greatest percentage of increments in KDML105(187%).Taken together,cultivation of rice in the slightly saline field did not alter its eating and cooking qualities,while enhanced some nutritional properties such as proteins,minerals,and secondary metabolites like phenolic compounds.

Starch Digestibility and Physicochemical and Cooking Properties of Irradiated Rice Grains

This study evaluated the starch digestibility, physicochemical properties and cooking characteristics of polished rice（varieties IRGA417 and IAC202） subjected to the doses of 0（as the control）, 1, 2 and 5 k Gy of gamma radiation. The highest dose decreased the apparent amylose content, peak viscosity, water absorption and volume expansion. Irradiation increased the solid loss by 119% and 187% for IAC202 and IRGA417, respectively, when comparing the higher dose with the control. For IAC202, irradiation decreased the rapidly digestible starch and increased the slowly digestible starch（SDS） and resistant starch（RS）. IRGA417 showed an elevation of SDS and a reduction in RS. And 1 k Gy dose of gamma radiation generated the highest level of RS for both the two varieties and presented the smallest changes in other physicochemical and cooking properties.

Comparison of Starch Granule Size Distribution Between Hard and Soft Wheat Cultivars in Eastern China

Granule size distribution of wheat starch is an important characteristic that can affect its chemical composition and functionality. Two types of wheat cultivars, the hard and soft wheat cultivars, grown at Tai＇an Experimental Station of Shandong Agricultural University, Taian, Shandong, China, were examined in this study. The granule size distribution and amylose contents in wheat grains were studied and compared, and relationships between the properties were identified. A clear bimodal distribution of granule size was shown in all wheat cultivars. Volume distribution of starch granules shows the typical bimodal with peak values in the ranges of 5.6-6.1μm and 20.7-24.9μm, respectively. Also, granule surface area distribution was bimodal with peak values in the ranges of 2.4-3.2μm and 20.7-24.9μm, respectively. Number distribution of granules was a typical population with a peak value in the range of 0.54-1.05μm. Contributions from the granules 〈 2.8μm and 〈 9.9μm to the total volume were in the ranges of 94.2-95.1% and 99.7-99.9% of total number, respectively. Proportions of granules〈2.8μm, 2.8-9.9μm, 9.9-22.8μm, and 22.8-42.8μm were in the ranges of 12.9-14.3%, 28.4-31.1%, 33.5-35.6%, and 19.7-22.7% for hard wheat, and 10.3-13.9%, 26.6-28.1%, 32.7-34.6%, and 24.2-27% for soft wheat. Hard wheat had greater B-type granules （ 〈 9.9μm）, and had fewer granules of 22.8-42.8μm than soft wheat. Amylose content was positively related to volume percentage of granules 22.8-42.8μm, and negatively related to volume percentage of granules 2.8-22.8μm.

Effects of Early- and Late-Sowing on Starch Accumulation and Associated Enzyme Activities During Grain Filling Stage in Rice

The environmental temperature occurring during the grain filling stage is an important factoraffecting starch synthesis and accumulation in rice. We investigated starch accumulation, amylaseactivity and starch granule size distribution in two low-amylose japonica rice varieties, Nanjing 9108 andFujing 1606, grown in the field at different filling temperatures by manipulating sowing date. The two ricevarieties exhibited similar performances between two sowing dates. Total starch, amylose andamylopectin contents were lower at the early-filling stage of T1 treatment (Early-sowing) compared withthose at the same stage in T2 treatment (Late-sowing). In contrast, at the late-filling stage, when fieldtemperatures were generally decreasing, total starch and amylopectin contents in T1 were highercompared to those in T2. The ideal temperature for strong activity of ADP-glucose pyrophosphorylaseand soluble starch synthase was about 22℃. A higher temperature from the heading to maturity stagesin T1 increased the activities of starch branching enzyme and suppressed the activities of granule boundstarch synthetase and starch debranching enzyme. We found that rice produced larger-sized starchgranules under the T1 treatment. These results suggested that due to the early-sowing date, the hightemperature (30℃) occurring at the early-filling stage hindered starch synthesis and accumulation,however, the lower temperatures (22 ℃) at the late-filling stage allowed starch synthesis and accumulationto return to normal levels.

The Effect of High Temperature after Anthesis on Rice Quality and Starch Granule Structure of Endosperm

To ascertain the effect mechanism of high temperature after anthesis on rice quality, the experiment was conducted with two rice lines, the heat-tolerant line 996 and heat-sensitive line 4628, with high temperature and optimal temperature in the growth chamber to investigate the effect of high temperature stress after anthesis on rice appearance quality, milling quality, cooking and eating quality and starch granule structure of endo- sperm. The result showed that milled rice rate, head rice rate, amylose content and gel consistency of both lines decreased under high temperature stress after anthesis, while the ratio of grain length to width, chalky rate, chalkiness, protein content increased. Under high temperature treatments, gelatinization temperature, final viscosity, set back and peak time increased, breakdown decreased, Mg content and K content increased, Mg/K ra- tio decreased. Under same treatment, the extent of rice quality of heat tolerant line 996 affected by high temperature was lower than that of heat sensitive line 4628. Under high temperature stress after anthesis, starch granule arranged untightly, most of starch granules existed in the form of a single starch endosperm, refractive index decreased, transparency reduced, and led to the formation of chalk. Under high temperature stress af- ter anthesis, the increase of protein content, the decrease of Mg/K, the changes of rice RVA profile characteristics and starch granule structure of endosperm could be the main reason for the decrease of rice cooking and eating quality and appearance quality.

Effects of RNAi Silencing of SSIII Gene on Phosphorus Content and Characteristics of Starch in Potato Tubers

The sense and antisense fragments of the soluble starch synthase （SSII1） gene and the intron fragment of somatic embryogenesis receptor-like kinase （SERK1） gene were cloned from potato using PCR techniques. The RNAi plant expression vectors pBI-SSIII-RNAi and pBIC-SSIII-RNAi were constructed which containing fusion fragment of ＂sense fragment-intron-antisense fragment＂ driven by the constitutive expression promoter CaMV 35S and the tuber-specific expression promoter CIPP, respectively. The putative transgenic plants of potato cultivars Kexin-1 and Kexin-4 were obtained using Agrobacterium-mediated transformation method. PCR assay showed that the interference fragment of SSlll gene was integrated into potato genome. The RT-PCR analysis showed that the expression of SSlll gene was repressed apparently on the transcription level. Starch granules of the transgenic potato plants were different in morphology and became cracked in starch granule centre compared with the non-transgenic control plants. The amylose content of starch was increased by 2.68-29.05%, amylopectin to amylose ratio of starch had declined significantly, and the phosphorus content of the starch of the transgenic plants was reduced 9.94-58.36% compared with control plants. The results could provide certain foundation for improvement of potato starch quality.

Polyamines mediate the effect of post-anthesis soil drying on starch granule size distribution in wheat kernels

Polyamines(PAs) are important endogenous plant growth regulators responding to environmental stress and mediating many physiological processes including grain filling in cereals.This study investigated whether PAs mediate the effect of post-anthesis soil drying on starch granule size distribution,starch content,and weight of superior and inferior kernels of wheat(Triticum aestivum L.).Two wheat cultivars were grown in pots.Three treatments,well-watered(WW),moderate soil drying(MD) and severe soil drying(SD),were imposed from 9 days post-anthesis until maturity.PA levels in kernels and small,medium and large granules were measured.The results showed that superior kernels had much higher free spermidine(Spd) and free spermine(Spm) concentrations,larger volumes of medium starch granules,and smaller-sized large granules than did inferior kernels under all the treatments.Compared to WW,MD significantly increased the concentrations of free Spd and free Spm,activities of soluble starch synthase and granule-bound starch synthase,volume of medium granules,and starch content and kernel weight of inferior kernels,and decreased the size of large granules.SD produced the opposite effect.Application of Spd or Spm to spikes produced effects similar to those of MD,and application of an inhibitor of Spd and Spm synthesis produced effects similar to those of SD.These results suggest that PAs mediate the effect of post-anthesis soil drying on starch biosynthesis in wheat kernels by regulating key enzymes in starch synthesis and that elevated PA levels under MD increase the volume of medium granules and kernel weight of inferior kernels.

Amylose Content and Starch Granule Size in Rice Grains are Affected By Growing Season

Amylose content and starch granule size in grains influence rice quality,which differs between the early(ES)and late season(LS).The objective of this study was to determine the variation of amylose content and starch granule size between seasons and find the main reasons(e.g.,temperature and solar radiation)for the observed variation.Field experiments with six rice varieties(three high and three low amylose content rice)planted in the ES and LS were conducted in 2016 and 2017,respectively.The mean temperatures during the filling stage were higher in ES,however,the daily temperatures at 7-10 days after flowering(DAF)in 2016,and at 5-10,13-14 DAF in 2017 were higher in LS.The results showed that amylose content in LS was lower than in ES with high amylose content rice varieties(HACV);the opposite trend occurred with low amylose content rice varieties(LACV).The mean starch granule diameter was higher in LS than ES in 2016,but the opposite result occurred in 2017 with all rice.Our results suggest that higher temperatures increased and decreased the amylose content in HACV and LACV,respectively.Temperatures at 5-15 DAF were important for the formation of starch granules:lower temperatures during 10-14 DAF increased the proportion of larger starch granules(d>6.21μm,some with d>13.3μm),and higher temperatures at 5-6 DAF increased the proportion of starch granules with diameter 4.24-6.21μm.

Characterization of starch morphology, composition, physicochemical properties and gene expressions in oat

Starch is the major carbohydrate in oat （Avena sativa L.） and starch formation requires the coordinated actions of several synthesis enzymes. In this study, the granule morphology, composition and physicochemical properties of oat starch, as well as the expressions of starch synthesis genes were investigated during oat endosperm development. Under the scanning electron microscopy （SEM）, we observed that the unique compound granules were developed in oat endosperms at 10 days post anthesis （DPA） and then fragmented into irregular or polygonal simple granules from 12 DPA until seed maturity. The amylose content, branch chain length of degree of polymerization （DP=13-24）, gelatinization temperature and percentage of retrogradation were gradually increased during the endosperm development; whereas the distribution of short chains （DP=6-12） were gradually decreased. The relative expressions of 4 classes of 13 starch synthesis genes characterized in this study indicated that three expression pattern groups were significantly different among gene classes as well as among varied isoforms, in which the first group of starch synthesis genes may play a key role on the initiation of starch synthesis in oat endosperms.

A plastidic ATP/ADP transporter gene, IbAATP, increases starch and amylose contents and alters starch structure in transgenic sweetpotato

A plastidic adenosine triphosphate（ATP）/adenosine diphosphate（ADP） transporter（AATP） is responsible for importing ATP from the cytosol into plastids. In dicotyledonous plants, increasing ATP supply is a potential way to facilitate anabolic synthesis in heterotrophic plastids. In this study, a gene encoding the AATP protein, named Ib AATP, was isolated from sweetpotato（Ipomoea batatas（L.） Lam.）. Transcripts of Ib AATP were predominantly detected in the storage roots and leaves and were induced by exogenous sucrose and subjected to circadian rhythm. Transient expression of Ib AATP in tobacco and onion epidermal cells revealed the plastidic localization of Ib AATP. The overexpression of Ib AATP in sweetpotato significantly increased the starch and amylose contents and led to enlarged starch granules. The IbA ATP-overexpressing plants showed altered fine structure of amylopectin, which contained an increased proportion of chains with a degree of polymerization（DP） of 10–23 and a reduced number of chains with a DP of 5–9 and 24–40. In addition, starch from the transgenic plants exhibited different pasting properties. The transcript levels of starch biosynthetic genes, including Ib AGP, Ib GBSSI, Ib SSIIV, and Ib SBE, were differentially regulated in the transgenic plants. These results revealed the explicit role of Ib AATP in the starch biosynthesis of sweetpotato and indicated that this gene has the potential to be used to improve starch content and quality in sweetpotato and other plants.

Effects of CO_2 Enrichment on Microstructure and Ultrastructure of Two Species of Freshwater Green Algae

order to investigate the morphological response of freshwater green algae to elevated CO2 concentration, Chlamydomonas reinhardtii Dang and Scenedesmus obliquus Kutz were cultured with enriched CO2, and their microstructure and ultrastructure were examined by microscopy and electron microscopy. The effect of CO2 enrichment to 186 mumol/L, was insignificant on the shape and size of C. reinhardtii, but significant in reducing the volume of S. obliquus. High-CO2 increased the amount of chloroplast. The pyrenoids occurred in low-CO2-grown cells but not in high-CO2-grown ones and more starch granules were observed in the former.

Application of moderate nitrogen levels alleviates yield loss and grain quality deterioration caused by post-silking heat stress in fresh waxy maize

High temperature(HT)during grain filling is one of the most important environmental factors limiting maize yield and grain quality.Nitrogen(N)fertilizer is essential for maintaining normal plant growth and defense against environmental stresses.The effects of three N rates and two temperature regimes on the grain yield and quality of fresh waxy maize were studied using the hybrids Suyunuo 5(SYN5)and Yunuo 7(YN7)as materials.N application rates were 1.5,4.5,and 7.5 g plant-1,representing low,moderate,and high N levels(LN,MN,and HN,respectively).Mean day/night temperatures during the grain filling of spring-and summer-sown plants were 27.6/21.0°C and 28.6/20.0°C for ambient temperature(AT)and 35/21.0°C and 35/20.0°C for HT,respectively.On average,HT reduced kernel number,weight,yield,and moisture content by 29.8%,17.9%,38.7%,and 3.3%,respectively.Kernel number,weight,yield,moisture,and starch contents were highest under MN among the three N rates under both temperature regimes.HT reduced grain starch content at all N levels.HT increased grain protein content,which gradually increased with N rate.Mean starch granule size under MN was larger(10.9μm)than that under LN and HN(both 10.4μm)at AT.However,the mean size of starch granules was higher under LN(11.7μm)and lower under MN(11.2μm)at HT.Iodine binding capacity(IBC)was lowest under MN and highest under HN among the three N levels under both temperature regimes.In general,IBC at all N rates was increased by HT.Peak viscosity(PV)was gradually reduced with increasing N rate at AT.In comparison with LN,PV was increased by MN and decreased by HN at HT.Retrogradation percentage gradually increased with N rate at AT,but was lowest under MN among the three N rates at HT.LN+AT and MN+HT produced grain with high pasting viscosity and low retrogradation tendency.MN application could alleviate the negative effects of HT on the grain yield and quality of fresh waxy maize.

Chalkiness Characters and Scanning Electron Microscope Observation of Rice Grain Endosperm of Japonica Varieties in Southern Henan

Chalkiness characters affect not only the grain appearance,milling,eating and cooking qualities but also the grain nutritional quality in rice,thus it is one of the most important traits in rice. It is very important for us to investigate the relation of the chalkiness formation and the development of endosperm structure and starch granule of different rice varieties. Here,we have investigated the chalkiness characters such as chalkiness rate,chalkiness degree and chalkiness area in 15 japonica rice varieties from southern Henan. Furthermore,the endosperm structure and starch granules of rice grain were also observed with scanning electron microscope. The results showed that the 15 japonica varieties have a significantly linear relationship between the chalkiness rate and chalkiness degree. Among the varieties,the biggest difference is the chalkiness rate,the second is the chalkiness area,and the last is the chalkiness degree. Moreover,there is a certain correlation between the distribution of starch granules,the arrangement of endosperm cells and the occurrence of grain chalkiness in the different rice varieties. For the same variety,the starch granules of chalky and non-chalky grains have obvious difference,while the starch granules from the transparent part of chalky rice and non-chalky rice do not have significant difference. The results would provide useful references for the improvement of grain quality in rice.

Effects of CO_2 enrichment and spikelet removal on rice quality under open-air field conditions

The increase of atmospheric carbon dioxide（CO_2） concentration adversely affect several quality traits of rice grains, but the biochemical mechanism remains unclear. The objectives of this study were to determine how changes in the source-sink relationship affected rice quality. Source-sink manipulation was achieved by free-air CO_2 enrichment from tillering to maturity and partial removal of spikelet at anthesis using a japonica rice cultivar Wuyunjing 23. Enrichment with CO_2 decreased the head rice percentage and protein concentration of milled rice, but increased the grain chalkiness. In contrast, spikelet removal resulted in a dramatic increase in the head rice percentage and protein concentration, and much less grain chalkiness. Neither CO_2 enrichment nor spikelet removal affected the starch content, but the distribution of starch granule size showed distinct treatment effects. O n average, spikelet removal decreased the percentage of starch granules of diameter 〉10 and 5–10 μm by 23.6 and 5.6%, respectively, and increased those with a diameter of 2–5 and 〈2 μm by 4.6 and 3.3%, respectively. In contrast, CO_2 elevation showed an opposite response： increasing the proportion of large starch granules（〉5 μm） and decreasing that of 〈5 μm. The starch pasting properties were affected by spikelet removal much more than by CO_2 elevation. These results indicated that the protein concentration and starch granule size played a role in chalkiness formation under these experimental conditions.