Mature Grains Research Articles

Element distribution and iron speciation in mature wheat grains (Triticum aestivum L.) using synchrotron X-ray fluorescence microscopy mapping and X-ray absorption near-edge structure (XANES) imaging.

Several studies have suggested that the majority of iron (Fe) and zinc (Zn) in wheat grains are associated with phytate, but a nuanced approach to unravel important tissue-level variation in element speciation within the grain is lacking. Here, we present spatially resolved Fe-speciation data obtained directly from different grain tissues using the newly developed synchrotron-based technique of X-ray absorption near-edge spectroscopy imaging, coupling this with high-definition μ-X-ray fluorescence microscopy to map the co-localization of essential elements. In the aleurone, phosphorus (P) is co-localized with Fe and Zn, and X-ray absorption near-edge structure imaging confirmed that Fe is chelated by phytate in this tissue layer. In the crease tissues, Zn is also positively related to P distribution, albeit less so than in the aleurone. Speciation analysis suggests that Fe is bound to nicotianamine rather than phytate in the nucellar projection, and that more complex Fe structures may also be present. In the embryo, high Zn concentrations are present in the root and shoot primordium, co-occurring with sulfur and presumably bound to thiol groups. Overall, Fe is mainly concentrated in the scutellum and co-localized with P. This high resolution imaging and speciation analysis reveals the complexity of the physiological processes responsible for element accumulation and bioaccessibility.

Seed dormancy QTL identification across a Sorghum bicolor segregating population

Pre-harvest sprouting (PHS) in Sorghum bicolor is one of the main constrains for its production in the central region of Argentina, as grain maturation often coincides with rainy or high environmental humidity conditions. The obtention of more dormant genotypes with higher PHS resistance has always been a desirable trait for breeders but the typical quantitative nature of seed dormancy makes its manipulation difficult through classical breeding. Dissecting this quantitative variability into quantitative trait loci (QTL) is a main concern especially in cereal species. In this work, a sorghum segregating population including 190 families was genotyped with microsatellite markers and the SbABI5 candidate gene. A genetic map encompassing 96 markers and a total length of 1331 cM was built. Seed dormancy was phenotyped in F3 and F4 panicles in two contrasting Argentinean environments (Castelar and Manfredi). Six seed dormancy QTL for mature grains were identified (qGI-1, qGI-3, qGI-4, qGI-6, qGI-7 and qGI-9) with the aid of QTL Cartographer and QTLNetwork, three of them (qGI-3, qGI-7 and qGI-9) being co-localised by both approaches. No epistasis was detected for the identified QTL but QTL-by-environment interaction was significant for qGI-7 and qGI-9. Interestingly, seed dormancy candidate genes SbABI3/VP1 and SbGA20ox3 were located within qGI-3, which makes them noteworthy candidate genes for this QTL.

Early assessment of the yield loss in rice due to the brown planthopper using a hyperspectral remote sensing method

ABSTRACTThe brown planthopper (BPH) is an important pest in rice. Rice losses due to BPH's damage are often more than 10% of yield. Assessments on loss rates from BPH are now basically dependent on experiential indices. However, early assessments on rice yield losses using hyperspectral data are still rare. In this study, reflectance from rice canopy was measured in net cages after different densities of BPH release. Results showed that reflectance in the near-infrared region (750–1000 nm) from milk grain stage, and in 400–531 nm and 567–705 nm from mature grain stage was closely related to BPH density. These spectral indices: red-edge parameters (λr, Dλr, Sλr), ratio of the maximal reflectance in green (490–560 nm) to minimal reflectance (640–740 nm) in red (RGREEN/RRED), ratio of the near-infrared peak (RNIR) to RRED, normalized difference vegetation index between RNIR and RRED and soil-adjusted vegetation index (SAVI) from both milk and mature grain stages, indicated BPH densities well. As expected, rate of loss in rice grain was higher with increasing BPH density. SAVI, Dλr, Sλr and reflectance at 760 nm (R760) from milk grain stage were significantly correlated with rates of loss in panicle, and 1000-grain weight and multiple-linear regression models for detecting loss rate were established. The hyperspectral reflectance from rice at milk grain stage can be used to assess rice yield losses due to BPH and improve management policies.

Comparative Proteomic Analysis of Two Barley Cultivars (Hordeum vulgare L.) with Contrasting Grain Protein Content.

Grain protein contents (GPCs) of barley seeds are significantly different between feed and malting barley cultivars. However, there is still no insight into the proteomic analysis of seed proteins between feed and malting barley cultivars. Also, the genetic control of barley GPC is still unclear. GPCs were measured between mature grains of Yangsimai 3 and Naso Nijo. A proteome profiling of differentially expressed protein was established by using a combination of 2-DE and tandem mass spectrometry. In total, 502 reproducible protein spots in barley seed proteome were detected with a pH range of 4–7 and 6–11, among these 41 protein spots (8.17%) were detected differentially expressed between Yangsimai 3 and Naso Nijo. Thirty-four protein spots corresponding to 23 different proteins were identified, which were grouped into eight categories, including stress, protein degradation and post-translational modification, development, cell, signaling, glycolysis, starch metabolism, and other functions. Among the identified proteins, enolase (spot 274) and small subunit of ADP-glucose pyrophosphorylase (spot 271) are exclusively expressed in barley Yangsimai 3, which may be involved in regulating seed protein expression. In addition, malting quality is characterized by an accumulation of serpin protein, Alpha-amylase/trypsin inhibitor CMb and Alpha-amylase inhibitor BDAI-1. Most noticeably, globulin, an important storage protein in barley seed, undergoes post-translational processing in both cultivars, and also displays different expression patterns.

Quantitative Trait Loci and Inter-Organ Partitioning for Essential Metal and Toxic Analogue Accumulation in Barley.

The concentrations of both essential nutrients and chemically similar toxic analogues accumulated in cereal grains have a major impact on the nutritional quality and safety of crops. Naturally occurring genetic diversity can be exploited for the breeding of improved varieties through introgression lines (ILs). In this study, multi-element analysis was conducted on vegetative leaves, senesced flag leaves and mature grains of a set of 54 ILs of the wild ancestral Hordeum vulgare ssp. spontaneum in the cultivated variety Hordeum vulgare ssp. vulgare cv. Scarlett. Plants were cultivated on an anthropogenically heavy metal-contaminated soil collected in an agricultural field, thus allowing simultaneous localization of quantitative trait loci (QTL) for the accumulation of both essential nutrients and toxic trace elements in barley as a model cereal crop. For accumulation of the micronutrients Fe and Zn and the interfering toxin Cd, we identified 25, 16 and 5 QTL, respectively. By examining the gene content of the introgressions, we associated QTL with candidate genes based on homology to known metal homeostasis genes of Arabidopsis and rice. Global comparative analyses suggested the preferential remobilization of Cu and Fe, over Cd, from the flag leaf to developing grains. Our data identifies grain micronutrient filling as a regulated and nutrient-specific process, which operates differently from vegetative micronutrient homoeostasis. In summary, this study provides novel QTL for micronutrient accumulation in the presence of toxic analogues and supports a higher degree of metal specificity of trace element partitioning during grain filling in barley than previously reported for other cereals.

Interruption of magnesium supply at heading influenced proteome of peripheral layers and reduced grain dry weight of two wheat (Triticum aestivum L.) genotypes

Magnesium (Mg), an indispensable mineral for plant growth, is concentrated in the peripheral layers (PLs) of the mature grain of wheat. The supply of Mg was interrupted from plant heading to maturity and a proteomic approach was used to investigate the PLs at three stages of development. Two genotypes with contrasting concentrations of Mg in the grain were studied: Apache (low Mg) and MgHL (high Mg). The concentration of Mg was significantly reduced in the roots (10–21%), straw (18–50%) and grain (24–10%), respectively. Mg deficiency altered enzymes involved in photosynthesis, glycolysis, respiration, amino acid synthesis, cell division, protein degradation and folding at early stages, especially in MgHL. This latter had smaller grain by reducing grain potential size and dry matter accumulation. By contrast in Apache, few proteins were affected at early stages and proteins related to stress/defense and arginine/proline metabolism were up accumulated resulting in lower number of grains per ear (24.9%). This study showed that Mg in PLs plays an important role in cell division, ATP generation, carbohydrate and amino acid metabolism, and hence may influence grain potential size and assimilates in grain, which determines grain weight. These results should help wheat breeders improve Mg content and hence grain yield. Biological significanceMagnesium (Mg) is an abundant cation and is involved in many cell activities. Its role in determining wheat productivity remains unclear. This study is the first to investigate how Mg deficiency influences the physiological characters of wheat and dry matter in the grain in two genotypes with contrasting Mg content. Moreover, Mg is concentrated in peripheral layers of grain, which are known to play a critical role in grain development. In this study, we investigated proteins in the peripheral layers expressed differentially in three development stages to identify the mechanism by which Mg influences grain development. This study revealed that the supply of Mg influences grain yield and that Mg regulates proteins related to cell metabolism and stress defense in grain.

Highly Sprouting-Tolerant Wheat Grain Exhibits Extreme Dormancy and Cold Imbibition-Resistant Accumulation of Abscisic Acid.

Pre-harvest sprouting (PHS) of wheat (Triticum aestivum L.) grains induces hydrolyzing enzymes such as α-amylase, which considerably decreases wheat product quality. PHS occurs when cool and wet weather conditions before harvest break dormancy and induce grain germination. In this study, we used PHS-tolerant varieties, Gifu-komugi (Gifu) and OS38, to characterize the mechanisms of both dormancy breakage and dormancy maintenance at low temperatures. Physiologically mature Gifu grains exhibited dormancy after imbibition at 20°C, but germinated at 15°C. In contrast, OS38 grains remained dormant even at temperatures as low as 5°C. Embryo half-grains cut out from the dormant Gifu grains germinated by imbibition at 20°C, similar to conventional varieties worldwide. However, OS38 embryo half-grains were still dormant. Hormonome and pharmacological analyses suggested that ABA and gibberellin metabolism are important for temperature-dependent dormancy maintenance and breakage. Imbibition at 15°C decreased ABA levels but increased gibberellin levels in embryos of freshly harvested Gifu grains. Additionally, low temperatures induced expression of the ABA catabolism genes,TaABA8' OH1 and TaABA8' OH2, and the gibberellin biosynthesis gene,TaGA3ox2, in the embryos. However, in embryos of freshly harvested OS38 grains, ABA levels were increased while gibberellin levels were suppressed at 15°C. In these dormant embryos, low temperatures induced the TaNCED ABA biosynthesis genes, but suppressed TaABA8' OH2 and TaGA3ox2.These results show that the regulatory mechanism influencing the expression of ABA and gibberellin metabolism genes may be critical for dormancy maintenance and breakage at low temperatures. Our findings should help improve PHS-resistant wheat breeding programs.

Proteomic analysis of mature barley grains from C-hordein antisense lines

Hordeins are the major storage proteins in barley grains and are responsible for their low nutritional quality. Previously, antisense C-hordein barley lines were generated and were shown to contain a more balanced amino acid composition and an altered storage protein profile. In the present study, a proteomic approach that combined two-dimensional gel electrophoresis (2-DE) and mass spectrometry was used to (1) identify the changes in the protein profile of non-storage proteins (salt soluble fraction) in antisense C-hordein barley lines (L1, L2 and L3) and (2) map the differentially expressed proteins compared to the non-transgenic control line (Hordeum vulgare cv. Golden Promise). Moreover, the changes in the proteins were correlated with the more balanced amino acid composition of these lines, with special attention to the lysine content. The results showed that suppression of C-hordein expression does not exclusively affect hordein synthesis and accumulation. The more balanced amino acid composition observed in the transgenic lines L1, L2 and L3 was an indirect result of the profound alterations in the patterns of the non-storage proteins. The observed changes included up-regulated expression of the proteins involved in stress and detoxification (L1), defence (L2 and L3), and storage globulins (L1, L2 and L3). To a lesser extent, the proteins involved in grain metabolism were also changed. Thus, the increased essential amino acids content results from changes in distinct protein sources among the three antisense C-hordein lines analyzed, although the up-regulated expression of lysine-rich proteins was consistently observed in all lines.

Increasing abscisic acid levels by immunomodulation in barley grains induces precocious maturation without changing grain composition.

Abscisic acid (ABA) accumulates in seeds during the transition to the seed filling phase. ABA triggers seed maturation, storage activity, and stress signalling and tolerance. Immunomodulation was used to alter the ABA status in barley grains, with the resulting transgenic caryopses responding to the anti-ABA antibody gene expression with increased accumulation of ABA. Calculation of free versus antibody-bound ABA reveals large excess of free ABA, increasing signficantly in caryopses from 10 days after fertilization. Metabolite and transcript profiling in anti-ABA grains expose triggered and enhanced ABA-functions such as transcriptional up-regulation of sucrose-to-starch metabolism, storage protein synthesis and ABA-related signal transduction. Thus, enhanced ABA during transition phases induces precocious maturation but negatively interferes with growth and development. Anti-ABA grains display broad constitutive gene induction related to biotic and abiotic stresses. Most of these genes are ABA- and/or stress-inducible, including alcohol and aldehyde dehydrogenases, peroxidases, chaperones, glutathione-S-transferase, drought- and salt-inducible proteins. Conclusively, ABA immunomodulation results in precocious ABA accumulation that generates an integrated response of stress and maturation. Repression of ABA signalling, occurring in anti-ABA grains, potentially antagonizes effects caused by overshooting production. Finally, mature grain weight and composition are unchanged in anti-ABA plants, although germination is somewhat delayed. This indicates that anti-ABA caryopses induce specific mechanisms to desensitize ABA signalling efficiently, which finally yields mature grains with nearly unchanged dry weight and composition. Such compensation implicates the enormous physiological and metabolic flexibilities of barley grains to adjust effects of unnaturally high ABA amounts in order to ensure and maintain proper grain development.

Characterization of an IAA-glucose hydrolase gene TaTGW6 associated with grain weight in common wheat (Triticum aestivum L.)

In rice, the TGW6 gene determines grain weight and encodes a protein with indole-3-acetic acid (IAA)-glucose hydrolase activity. Its homolog in wheat, TaTGW6, is considered as a candidate gene related to grain development. To amplify this gene, we designed primers based on a homologous conserved domain of the rice TGW6 gene. Sequence analysis indicated that TaTGW6 comprises only one exon, with 1656 bp in total and an open reading frame of 1035 bp. Three alleles at TaTGW6 locus detected by the primer pair TG23 were designated as TaTGW6-a, TaTGW6-b and TaTGW6-c, respectively. Compared with TaTGW6-a, TaTGW6-b had a 6-bp InDel at the position 170 downstream of initiation codon, and TaTGW6-c was a null mutant. Both TaTGW6-b and TaTGW6-c could significantly increase grain size and weight other than TaTGW6-a; however, the former two alleles showed a low frequency distribution in modern varieties. TaTGW6 was located on chromosome 4AL using a recombinant inbred line population and a set of Chinese Spring nullisomic-tetrasomic lines. It was linked to the SSR locus Xbarc1047 with a genetic distance of 6.62 cM and explained 15.8–21.0 % of phenotypic variation of grain weight in four environments. Association analysis using a natural population and Chinese wheat mini-core collections additionally validated the relationship of TaTGW6 with grain weight; the gene could explain 7.7–12.4 % of phenotypic variation in three environments. Quantitative real-time PCR revealed that TaTGW6-b showed relatively lower expression than TaTGW6-a in immature grain at 20 and 30 days post-anthesis and in mature grain. The low expression of TaTGW6 generally associated with low IAA content, but with high grain weight. The novel functional marker, designated as TG23, can be used for marker-assisted selection to improve grain weight in wheat and also provides insights into the regulatory mechanism underlying grain weight.

Chlorosis correction and agronomic biofortification in field peas through foliar application of iron fertilizers under Fe deficiency

ABSTRACTEffectiveness of different iron (Fe) foliar sprays for leaf chlorosis correction and grain Fe boosting was studied in field peas under Fe deficiency. No chlorophyll reduction was observed in Fe deficient plants treated with foliar sprays. EDDHA [ethylenediamine-N,N′-bis(2-hydroxyphenylacetic acid)] followed by FeSO4 (73.7 mg/l Fe) treated at the start of flowering was most responsive in correcting chlorosis and increasing shoot dry biomass in peas. Inductively coupled plasma-atomic emission spectroscopy data showed significant increase of Fe in grains while treated with all foliar sprays at the time of grain filling in Fe-deficient plants. Among them, FeSO4 (73.7 mg/l Fe) was the most efficient in biofortifying Fe in mature grain under Fe deficiency in peas. Results also pinpoint that flowering is a suitable time for applying foliar sprays to boost Fe in mature grains. Taken together, application of Fe foliar sprays facilitated both chlorosis correction and Fe boosting in peas and can be further used by breeders and farmers.

ОПТИМИЗАЦИЯ МЕТОДА ОЦЕНКИ МОРОЗОУСТОЙЧИВОСТИ СОРТОВ ОЗИМОГО ЯЧМЕНЯ ПО ГИГРОСКОПИЧНОСТИ ЗРЕЛОГО ЗЕРНА

ОПТИМИЗАЦИЯ МЕТОДА ОЦЕНКИ МОРОЗОУСТОЙЧИВОСТИ СОРТОВ ОЗИМОГО ЯЧМЕНЯ ПО ГИГРОСКОПИЧНОСТИ ЗРЕЛОГО ЗЕРНА

Antioxidant and protease-inhibitory potential of extracts from grains of oat

AbstractThe most of important crops cultivated for production of foods and feeds could be considered as plants possessing nutraceutical or medically interesting compounds, especially if can be eaten without processing. Chemical and biological parameters that were evaluated in 100 oat (Avena sativa L.) genotypes were others than those that are important in food and feed production. Contents of polyphenols and flavonoids, radical scavenging activity (DPPH), and inhibitory activities against five proteases (trypsin, thrombin, urokinase, elastase, cathepsin B) were analyzed in extracts from mature grains. The antioxidant activity (DPPH) correlated to the content of total polyphenols. Only a minority (15 from 100) of analyzed genotypes created separate subgroup with a high content of polyphenols, flavonoids, and high antioxidant activity. The best in these parameters were genotypes CDC-SOL-FI, Saul, and Avesta, respectively. Fifteen other genotypes assembled another minority subgroup (also 15 from 100) on the basis of their high inhibitory activities against tested proteases. The highest trypsin-, urokinase-, and elastase-inhibitory activities were in genotype Racoon, the best in thrombin-, and cathepsin B-inhibitory activities were genotypes Expression and SW Kerstin, respectively. Three oats genotypes – Rhea, AC Percy, and Detvan appeared in both subgroups.

Cover Crop Impact on Nitrogen Availability and Dry Bean in an Organic System

Cover crops could play an important role in increasing nutrient availability and crop yield in organic systems. This study had two main objectives, (i) compare the effects of medium red clover (Trifolium pratense L.), cereal rye (Secale cereale L.), and oilseed radish (Raphanus sativus L.) cover crops on N availability and dry bean (Phaseolus vulgaris L.) growth and yield and (ii) compare the use of ion exchange resin and chlorophyll content monitoring with traditional soil KCl extractions for determining soil inorganic N availability. Red clover increased soil inorganic N up to 55 kg ha−1, resulting in delayed maturity and greater grain N in some site‐years; population and yield were not affected. Rye reduced soil inorganic N and caused early maturity of bean in some site‐years; at maximum dry biomass production (12.8 Mg ha−1) rye reduced dry bean yield. Oilseed radish occasionally increased inorganic N and bean populations; yield was not influenced. Cover crop and weed C/N ratios and C and N content were the most consistent factors influencing soil inorganic N. Soil inorganic N measurements from soil extractions and resin strips were positively correlated, however the use of resin strips may be constrained due to frequent cultivation in organic systems. Correlations were seldom observed among these two direct N measures and dry bean relative chlorophyll content. Overall, planting cover crops did not benefit dry bean yield, however producers may be able to manipulate soil inorganic N by focusing on cover crop quality, depending on their goals and species selection.

Suitable growing zone and yield potential for late-maturity type of Yongyou japonica/indica hybrid rice in the lower reaches of Yangtze River, China

Late-maturity type of Yongyou japonica/indica hybrids series (LMYS) have shown great yield potential, and are being widely planted in the lower reaches of Yangtze River, China. Knowledge about suitable growing zone and evaluation of yield advantage is of practical importance for LMYS in this region. Fifteen LMYS, two high-yielding inbred japonica check varieties (CK-J) and two high-yielding hybrid indica check varieties (CK-I) were grown at Xinghua (119.57°E, 33.05°N) of Lixiahe region, Yangzhou (119.25°E, 32.30°N) of Yanjiang region, Changshu (120.46°E, 31.41°N) of Taihu Lake region, and Ningbo (121.31°E, 29.45°N) of Ningshao Plain in 2013 and 2014. The results showed that maturity dates of the 15 were later than the secure maturity date at Xinghua and 6, 14 and 15 LMYS were mature before the secure maturity date at Yangzhou, Changshu and Ningbo, respectively. One variety was identified as high-yielding variety among LMYS (HYYS) at Yangzhou, 8 HYYS in 2013 and 9 HYYS in 2014 at Changshu, 9 HYYS at Ningbo. HYYS here referred to the variety among LMYS that was mature before the secure maturity date and had at least 8% higher grain yield than both CK-J and CK-I at each experimental site. Grain yield of HYYS at each experimental site was about 12.0 t ha−1 or higher, and was significantly higher than CK varieties. High yield of HYYS was mainly attributed to larger sink size due to more spikelets per panicle. Plant height of HYYS was about 140 cm, and was significantly higher than check varieties. Significant positive correlations were recorded between duration from heading to maturity stage and grain yield, and also between whole growth periods and grain yield. HYYS had obvious advantage over check varieties in biomass accumulation and leaf area duration from heading to maturity stage. Comprehensive consideration about safe maturity and yield performance of LMYS at each experimental site, Taihu Lake region (representative site Changshu) and Ningshao Plain (representative site Ningbo) were thought suitable growing zones for LMYS in the lower reaches of Yangtze River. The main factors underlying high yield of HYYS were larger sink size, higher plant height, longer duration from heading to maturity stage and whole growth periods, and higher biomass accumulation and leaf area duration during grain filling stage.

Effects of Heat Stress on Metabolite Accumulation and Composition, and Nutritional Properties of Durum Wheat Grain.

Durum wheat (Triticum turgidum (L.) subsp. turgidum (L.) convar. durum (Desf.)) is momentous for human nutrition, and environmental stresses can strongly limit the expression of yield potential and affect the qualitative characteristics of the grain. The aim of this study was to determine how heat stress (five days at 37 °C) applied five days after flowering affects the nutritional composition, antioxidant capacity and metabolic profile of the grain of two durum wheat genotypes: “Primadur”, an elite cultivar with high yellow index, and “T1303”, an anthocyanin-rich purple cultivar. Qualitative traits and metabolite evaluation (by gas chromatography linked to mass spectrometry) were carried out on immature (14 days after flowering) and mature seeds. The effects of heat stress were genotype-dependent. Although some metabolites (e.g., sucrose, glycerol) increased in response to heat stress in both genotypes, clear differences were observed. Following the heat stress, there was a general increase in most of the analyzed metabolites in “Primadur”, with a general decrease in “T1303”. Heat shock applied early during seed development produced changes that were observed in immature seeds and also long-term effects that changed the qualitative and quantitative parameters of the mature grain. Therefore, short heat-stress treatments can affect the nutritional value of grain of different genotypes of durum wheat in different ways.

Biosynthesis of Essential Polyunsaturated Fatty Acids in Wheat Triggered by Expression of Artificial Gene.

The artificial gene D6D encoding the enzyme ∆6desaturase was designed and synthesized using the sequence of the same gene from the fungus Thamnidium elegans. The original start codon was replaced by the signal sequence derived from the wheat gene for high-molecular-weight glutenin subunit and the codon usage was completely changed for optimal expression in wheat. Synthesized artificial D6D gene was delivered into plants of the spring wheat line CY-45 and the gene itself, as well as transcribed D6D mRNA were confirmed in plants of T0 and T1 generations. The desired product of the wheat genetic modification by artificial D6D gene was the γ-linolenic acid. Its presence was confirmed in mature grains of transgenic wheat plants in the amount 0.04%–0.32% (v/v) of the total amount of fatty acids. Both newly synthesized γ-linolenic acid and stearidonic acid have been detected also in leaves, stems, roots, awns, paleas, rachillas, and immature grains of the T1 generation as well as in immature and mature grains of the T2 generation. Contents of γ-linolenic acid and stearidonic acid varied in range 0%–1.40% (v/v) and 0%–1.53% (v/v) from the total amount of fatty acids, respectively. This approach has opened the pathway of desaturation of fatty acids and production of essential polyunsaturated fatty acids in wheat.

Gel-free/label-free proteomic analysis of developing rice grains under heat stress

High temperature markedly reduces the yields and quality of rice grains. To identify the mechanisms underlying heat stress-induced responses in rice grains, proteomic technique was used. Developing Khao Dawk Mali 105 rice grains at the milky, dough, and mature stages were treated at 40°C for 3days. Aromatic compounds were decreased in rice grains under heat stress. The protein abundance involved in glycolysis and tricarboxylic acid cycle, including glyceraldehyde 3-phosphate dehydrogenase and citrate synthase, was changed in milky and dough grains after heat treatment; however, none changes in mature grains. The abundance involved in amino acid metabolism was increased in dough grains, but decreased in milky grains. In addition, the abundance involved in starch and sucrose metabolism, such as starch synthase, ADP-glucose pyrophosphorylase, granule-bound starch synthase, and alpha amylase, was decreased in milky grains, but increased in dough grains. A number of redox homeostasis-related proteins, such as ascorbate peroxidase and peroxiredoxin, were increased in developing rice grains treated with heat stress. These results suggest that in response to heat stress, the abundance of numerous proteins involved in redox homeostasis and carbohydrate biosynthetic pathways may play a major role in the development of KDML105 rice grains. Biological significanceYield of Khao Dawk Mali 105 rice, which is an economical aromatic rice, was disrupted by environmental stress. Rice grains developed under heat stress caused loss of aroma compound. To identify the mechanism of heat response in rice grain, gel-free/label-free proteomic technique was used. The abundance of proteins involved in glycolysis and tricarboxylic acid cycle was disrupted by heat stress. High temperature limited starch biosynthesis; however, it enhanced sugar biosynthesis in developing rice grains. Redox homeostasis related proteins were disrupted by heat stress. These results suggest that proteins involved in redox homeostasis and carbohydrate pathway might play a major role in developing grains in Khao Dawk Mali 105 rice under heat stress.

Comparative Expression Analysis of Metal Homeostasis-Related Genes in Rice Genotypes Differing in Grain Micronutrient Levels

To understand the role of metal homeostasis-related genes in rice, micronutrient levels of different tissue types were analysed at mid grain-filling stage followed by the expression analysis of candidate genes in these tissues. Subsequently, the association between the gene expression pattern and micronutrient level in tissues as well as mature grains was analysed. Out of 11 candidate genes used for gene expression analysis utilizing bulked cDNA based RT-PCR, 8 genes showed high level of expression in flag leaf and second leaf tissues. Four genes showed poor level of expression in immature grains and low to negligible expression in stem tissues. Further, six candidate genes were selected based on differential response of cDNA bulk analysis for the expression studies of individual rice genotypes, including four high-zinc rice genotypes, namely R-RHZ-LI-25, IR92970-111-1-2, R-RHZ-SM- 3, R-RHZ-SM-4 which showed higher level of expression for genes OsVIT1 , OsFER1 , OsYSL2 and OsYSL9 . Whereas low or negligible level gene expression in stem tissue of five genes, except OsFER1 shows that rice stem tissue could be involved in the uptake of micronutrients. The thorough characterization of genes in this study provides deeper insight into the tightly regulated mechanism of metal homeostasis with respect to different tissue types and understanding of source-sink relationship of mineral acquisition and remobilization.

Starch Debranching Enzyme Activity and Its Effects on Some Starch Physicochemical Characteristics in Developing Substituted and Complete Triticales (X Triticosecale Wittmack)

The present work was carried out to make a comparison between the starch debranching enzyme activity and the synthesis of total carbohydrates, starch, amylose, and amylopectin in developing kernels of two sets of triticales differing in genome composition (complete and substituted triticales). The results showed that the carbohydrate and starch accumulation observed in both genotypes of triticales increased rapidly at the early stage, reaching the maximum at the medium stage and decreasing slightly or remaining relatively constant at the last stage of kernel development. At the end of the development stage, the mature grains of complete and substituted triticales accumulated 62.5 and 54.6% starch, respectively. Both sets of triticales showed the same amylose accumulation profile as well as degree of polymerization; however, at the maturity stage there was a small but significant difference between the two sets. The final polymerization values of amylose chains in mature seeds of complete and substituted triticales were DP 1,549 and 1,313. A and B1 amylopectin chains of the complete triticales were approximately 19 and 33% shorter than those of the substituted triticales. The complete triticales showed higher isoamylase and pullulanase activities than the substituted triticales, which coincided with the highest starch and amylose contents observed in kernels under development. The results of this study demonstrate that the synthesis of starch differs between the triticale genotypes used in the study, which influences their chemical structure and physicochemical properties.