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Inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IPK1) catalyzes the final step in phytic acid (InsP6) synthesis. In this study, the effects of OsIPK1 mutations on InsP6 synthesis, grain filling and their underlying mechanisms were investigated. Seven gRNAs were designed to disrupt the OsIPK1 gene via CRISPR/CAS9 system. Only 4 of them generated 29 individual insertion or deletion T0 plants, in which nine biallelic or heterozygous genotypes were identified. Segregation analysis revealed that OsIPK1 frameshift mutants are homozygous lethality. The biallelic and heterozygous frameshift mutants exhibited significant reduction in yield-related traits, particularly in the seed-setting rate and yield per plant. Despite a notable decline in pollen viability, the male and female gametes had comparable transmission rates to their progenies in the mutants. A significant number of the filling-aborted (FA) grains was observed in mature grains of these heterozygous frameshift mutants. These grains exhibited a nearly complete blockage of InsP6 synthesis, resulting in a pronounced increase in Pi content. In contrast, a slight decline in InsP6 content was observed in the plump grains. During the filling stage, owing to the excessive accumulation of Pi, starch synthesis was significantly impaired, and the endosperm development-specific gene expression was nearly abolished. Consistently, the activity of whereas AGPase, a key enzyme in starch synthesis, was significantly decreased and Pi transporter gene expression was upregulated in the FA grains. Taken together, these results demonstrate that OsIPK1 frameshift mutations result in excessive Pi accumulation, decreased starch synthesis, and ultimately leading to lower yields in rice.Key message: OsIPK1 frameshift mutation results in excessive Pi accumulation in the filling-aborted grain and reduced grain yield in rice. [ABSTRACT FROM AUTHOR]

SUMMARY: Understanding and optimizing the process of grain filling helps the quest to maximize rice (Oryza sativa L.) seed yield and quality, yet the intricate mechanisms at play remain fragmented. Transcription factors (TFs) are major players in the gene networks underlying the grain filling process. Here, we employed grain incomplete filling (OsGIF1)/cell wall invertase 2, a key gene involved in grain filling, to explore its upstream TFs and identified a bZIP family TF, OsbZIP10, to be a transcriptional activator of OsGIF1. Rice grains of the knockouts of OsbZIP10 showed increased white‐core rates but lower amylose content (AC), leading to better eating and cooking qualities in all genetic backgrounds investigated, though the impact of mutations in OsbZIP10 on grain weight depended on genetic background. Multi‐omics analyses suggested that, in addition to OsGIF1, multiple genes involved in different biological processes contributing to grain filling were targeted by OsbZIP10, including OsAGPS1, a gene encoding the ADP‐Glc pyrophosphorylase (AGPase) small subunit, and genes contributing to homeostasis of reactive oxygen species. Distinct genetic make‐up was observed in OsbZIP10 between japonica and indica rice varieties, with the majority varieties of each subspecies belonging to two different haplotypes that were closely associated with AC. Overexpressing the haplotype linked to high‐AC in the low‐AC genetic background increased AC. Overall, this study sheds crucial light on the significance of the OsbZIP10–OsGIF1 module in the determination of rice grain quality, offering a potential avenue for genetic engineering of rice to produce seeds with tailored attributes. [ABSTRACT FROM AUTHOR]

Balancing the ATP: NADPH demand from plant metabolism with supply from photosynthesis is essential for preventing photodamage and operating efficiently, so understanding its drivers is important for integrating metabolism with the light reactions of photosynthesis and for bioengineering efforts that may radically change this demand. It is often assumed that the C3 cycle and photorespiration consume the largest amount of ATP and reductant in illuminated leaves and as a result mostly determine the ATP: NADPH demand. However, the quantitative extent to which other energy consuming metabolic processes contribute in large ways to overall ATP: NADPH demand remains unknown. Here, we used the metabolic flux networks of numerous recently published isotopically non-stationary metabolic flux analyses (INST-MFA) to evaluate flux through the C3 cycle, photorespiration, the oxidative pentose phosphate pathway, the tricarboxylic acid cycle, and starch/sucrose synthesis and characterize broad trends in the demand of energy across different pathways and compartments as well as in the overall ATP:NADPH demand. These data sets include a variety of species including Arabidopsis thaliana, Nicotiana tabacum, and Camelina sativa as well as varying environmental factors including high/low light, day length, and photorespiratory levels. Examining these datasets in aggregate reveals that ultimately the bulk of the energy flux occurred in the C3 cycle and photorespiration, however, the energy demand from these pathways did not determine the ATP: NADPH demand alone. Instead, a notable contribution was revealed from starch and sucrose synthesis which might counterbalance photorespiratory demand and result in fewer adjustments in mechanisms which balance the ATP deficit. [ABSTRACT FROM AUTHOR]

The storage substances of rice are critical factors that simultaneously affect grain yield and quality. Starch is the major component in the endosperm, directly affecting the seed yield as well as the cooking and eating quality of grains. Growing evidence suggests that some transcription factors are involved in regulating seed traits and starch biosynthesis. Here, we reveal the role of an NF-Y transcription factor gene, OsNF-YA8 , in regulation of seed traits and starch accumulation. OsNF-YA8 encodes a rice NF-YA protein containing the 166-amino acid. The RT-qPCR showed that OsNF-YA8 is expressed in roots, stems, leaves, flag leaves, and panicles, and dramatically increased in panicle during rice filling. Intracellular localization analysis showed that OsNF-YA8 is a nucleus-localized protein. In the OsNF-YA8 overexpression lines, rice grain length increased by 13.80–14.21 %, rice grain thickness decreased by 3.22–6.65 %, and the total starch content increased by 14.70–33.14 %. The overexpression of the OsNF-YA8 also decreased average starch granule size, and changed starch properties. Moreover, OsNF-YA8-overexpression up-regulated the expressions of genes involved in sucrose synthesis/transport and starch synthesis. By contrast, RNAi of OsNF-YA8 decreased the grain length, grain width, 1000 grain weight and starch content of rice seeds, and down-regulated the expressions of genes involved in sucrose synthesis/transport and starch synthesis. These results suggest that OsNF-YA8 enhances starch accumulation and confers seed traits by positively regulating starch biosynthesis in rice. The findings provide a new perspective on genetic improvement of crop agronomic traits and seed quality. [Display omitted] • OsNF-YA8 encodes a nucleus-localized protein with 166-amino acid. • The overexpression of the OsNF-YA8 confers seed improvement traits. • OsNF-YA8 promotes starch accumulation by positively regulates starch biosynthesis. • OsNF-YA8 impacts on rice grain size and physicochemical properties of starch grains. • OsNF-YA8 upregulates the expression of sucrose and starch synthesis-related genes. [ABSTRACT FROM AUTHOR]

(1) The development of sweet potato storage roots is impacted by nitrogen (N) levels, with excessive nitrogen often impeding development. Starch synthesis enzymes such as sucrose synthase (SUS) and ADP-glucose pyrophosphorylase (AGPase) are pivotal in this context. Although the effects of excessive nitrogen on the formation of sweet potato storage roots are well documented, the specific responses of IbSUSs and IbAGPases have not been extensively reported on. (2) Pot experiments were conducted using the sweet potato cultivar "Pushu 32" at moderate (MN, 120 kg N ha−1) and excessive nitrogen levels (EN, 240 kg N ha−1). (3) Nine IbSUS and nine IbAGPase genes were categorized into three and two distinct subgroups based on phylogenetic analysis. Excessive nitrogen significantly (p < 0.05) suppressed the expression of IbAGPL1, IbAGPL2, IbAGPL4, IbAGPL5, IbAGPL6, IbAGPS1, and IbAGPS2 in fibrous roots and IbSUS2, IbSUS6, IbSUS7, IbSUS8, IbSUS9, IbAGPL2, and IbAGPL4 in storage roots, and then significantly (p < 0.05) decreased the SUS and AGPase activities and starch content of fibrous root and storage root, ultimately reducing the storage root formation of sweet potato. Excessive nitrogen extremely significantly (p < 0.01) enhanced the expression of IbAGPL3, which was strongly negatively correlated with the number and weight of storage roots per plant. (4) IbAGPL3 may be a key gene in the response to excessive nitrogen stress and modifying starch synthesis in sweet potato. [ABSTRACT FROM AUTHOR]

Abstract Grain-filling of rice spikelets (particularly for the later flowering inferior spikelets) is an important characteristic that affects both quality and yield. Rice ratooning technology is used to cultivate a second crop from dormant buds that sprout from stubble left after the first harvest. This study used two rice varieties, the conventional indica rice ‘Jinhui 809’ and the hybrid indica-japonica rice ‘Yongyou 1540’, to assess the impact of rice ratooning on grain-filling. The results indicated that the grain-filling process in inferior spikelets of ratoon season rice (ISR) showed significant improvement compared to inferior spikelets of main crop (late season) rice (ISL). This improvement was evident in the earlier onset of rapid grain-filling, higher seed-setting percentage, and improved grain quality. A label-free quantitative proteomic analysis using mass spectrometry identified 1724 proteins with significant abundance changes, shedding light on the molecular mechanisms behind the improved grain-filling in ISR. The functional analysis of these proteins indicated that ratooning stimulated the metabolic processes of sucrose-starch, trehalose, and hormones in rice inferior spikelets, leading to enhanced enzyme activities related to starch synthesis, elevated concentrations of trehalose-6-phosphate (T6P), indole-3-acetic acid (IAA) and zeatin riboside (ZR) during the active grain-filling phase. This research highlighted the importance of the GF14f protein as a key regulator in the grain-filling process of ISR. It revealed that GF14f transcriptional and protein levels declined more rapidly in ISR compared to ISL during grain-filling. Additionally, the GF14f-RNAi plants specific to the endosperm exhibited improved quality in inferior spikelets. These findings suggest that the enhancement of starch synthesis, increased levels of IAA, ZR, and T6P, along with the rapid decrease in GF14f protein, play a role in enhancing grain-filling in ratoon season rice.

Grain size is one of the critical factors determining rice yield. Previous studies have found the grain-size-regulating function of Gα in rice. However, the regulatory mechanism underlying the development of rice grain mediated by Gα is still unclear. To reveal the functional mechanism of Gα in grain size regulation, a mutant of Gα (Gα-Cas9) was firstly constructed through a CRISPR/Cas9 strategy and was then grown in a greenhouse and field. The results showed that the seed length, plant height, 1000-grain weight, and spike length were significantly decreased in Gα-Cas9 compared to wild-type (WT) Pi-4b. During the grain filling stage, the increase in the grain dry weight of Pi-4b occurred earlier than that of Gα-Cas9. The total starch content and amylose content of matured grains of Pi-4b were higher than those of Gα-Cas9. Secondly, transcriptome sequencing analysis of Gα-Cas9 and Pi-4b during grain filling was performed to elucidate the functional pathways regulated by Gα. In total, 2867 and 4534 differentially expressed genes (DEGs) were discovered at 5 DAF and 10 DAF, and the starch and sucrose metabolism pathway enriched by DEGs was involved in grain size regulation mediated by Gα. Gα regulated the expression of starch-synthesis-related genes during grain filling, and the Gα protein interacted with OsNYC4 to trigger the sugar signaling pathway to promote starch accumulation in grain. Additionally, the GW2-WG1-OsbZIP47 pathway was switched off by Gα to relieve the inhibition of rice grain development. In this study, the results should provide new insights into the G protein signal transduction pathway. [ABSTRACT FROM AUTHOR]

This study aims to explore the formation mechanism of starch structure and the relationships between the appearance quality and starch structure of soft rice under different nitrogen levels. We comprehensively investigated the physiological aspects, starch structure variations, and appearance quality of soft rice in response to different nitrogen applications. The results showed that under the moderate nitrogen application (270 N), the soft rice exhibited the highest AGPase activity, the highest large‐starch granule content, and the lowest chalkiness. Under the highest nitrogen application (360 N), the soft rice exhibited the highest GBSS and DBE activity and the lowest SBE activity, the highest content of long‐branched amylopectin, the lowest relative crystallinity, the fewest ordered structures, the most amorphous structures, the largest semi‐crystalline lamellar thicknesses, and the highest transparency of chalk‐free rice. In conclusion, moderate nitrogen fertilization (270 N) improved the AGPase activity, which leaded to fuller starch granules and more compact endosperm in soft rice. Thus, the grain chalkiness of soft rice decreased. Continuous nitrogen application (0‐360 N) constantly increased the GBSS and DBE activity and reduced the SBE activity in soft rice, leading a lower content of short‐branched amylopectin and a higher content of long‐branched amylopectin in soft rice. Thus, the relative crystallinity and ordered structures of soft rice were reduced. These structures improved the transparency phenotype of soft rice. [ABSTRACT FROM AUTHOR]

Soluble starch synthases (SSs) play important roles in the synthesis of cassava starch. However, the expression characteristics of the cassava SSs genes have not been elucidated. In this study, the MeSSIII-1 gene and its promoter, from SC8 cassava cultivars, were respectively isolated by PCR amplification. MeSSIII-1 protein was localized to the chloroplasts. qRT-PCR analysis revealed that the MeSSIII-1 gene was expressed in almost all tissues tested, and the expression in mature leaves was 18.9 times more than that in tuber roots. MeSSIII-1 expression was induced by methyljasmonate (MeJA), abscisic acid (ABA), and ethylene (ET) hormones in cassava. MeSSIII-1 expression patterns were further confirmed in proMeSSIII-1 transgenic cassava. The promoter deletion analysis showed that the −264 bp to −1 bp MeSSIII-1 promoter has basal activity. The range from −1228 bp to −987 bp and −488 bp to −264 bp significantly enhance promoter activity. The regions from −987 bp to −747 bp and −747 bp to −488 bp have repressive activity. These findings will provide an important reference for research on the potential function and transcriptional regulation mechanisms of the MeSSIII-1 gene and for further in-depth exploration of the regulatory network of its internal functional elements. [ABSTRACT FROM AUTHOR]

Starch is a major storage carbohydrate in plants and is critical in crop yield and quality. Starch synthesis is intricately regulated by internal metabolic processes and external environmental cues; however, the precise molecular mechanisms governing this process remain largely unknown. In this study, we revealed that high red to far‐red (high R:FR) light significantly induces the synthesis of leaf starch and the expression of synthesis‐related genes, whereas low R:FR light suppress these processes. Arabidopsis phytochrome B (phyB), the primary R and FR photoreceptor, was identified as a critical positive regulator in this process. Downstream of phyB, basic leucine zipper transcription factor ELONGATED HYPOCOTYL5 (HY5) was found to enhance starch synthesis, whereas the basic helix‐loop‐helix transcription factors PHYTOCHROME INTERACTING FACTORs (PIF3, PIF4, and PIF5) inhibit starch synthesis in Arabidopsis leaves. Notably, HY5 and PIFs directly compete for binding to a shared G‐box cis‐element in the promoter region of genes encoding starch synthases GBSS, SS3, and SS4, which leads to antagonistic regulation of their expression and, consequently, starch synthesis. Our findings highlight the vital role of phyB in enhancing starch synthesis by stabilizing HY5 and facilitating PIFs degradation under high R:FR light conditions. Conversely, under low R:FR light, PIFs predominantly inhibit starch synthesis. This study provides insight into the physiological and molecular functions of phyB and its downstream transcription factors HY5 and PIFs in starch synthesis regulation, shedding light on the regulatory mechanism by which plants synchronize dynamic light signals with metabolic cues to module starch synthesis. [ABSTRACT FROM AUTHOR]

Although soybean and chickpea belong to the legume family, their seed starch content is very different. Currently, many studies focus on the molecular mechanisms of starch synthesis within a single species. However, the key genes and regulatory relationships responsible for the difference in seed starch content between the two species remain unknown. To elucidate the molecular mechanisms responsible for the above difference, multi-omics and bioinformatics analyses were used here to analyze gene expression patterns, protein–protein interaction networks, gene-transcription factor co-expression networks, and miRNA–gene regulatory relationships based on reported sucrose and starch metabolic genes in Arabidopsis. The results were as follows. First, seven differential expression genes of the two species in starch synthesis metabolism, including GBSS1, APL3, APS1, SS2, PTST, SBE2, and ISA, and the starch degradation gene BMY in soybean and chickpea, may contribute to their seed starch content differences. Then, the protein–protein interaction between DPEs and PHS may facilitate seed starch synthesis in chickpea. Finally, the positive regulation of two starch degradation genes (GmBMY and GmPHS) and four sucrose metabolism genes (GmHXK, GmPFK, GmTPS, and GmFRK) by transcription factors may lead to lower seed starch content in soybean. This study elucidates the possible molecular mechanisms underlying the difference in seed starch content between the two species and addresses the scientific problem of why soybean seeds have lower starch content than chickpea seeds. [ABSTRACT FROM AUTHOR]

Abstract This study aims to explore the formation mechanism of starch structure and the relationships between the appearance quality and starch structure of soft rice under different nitrogen levels. We comprehensively investigated the physiological aspects, starch structure variations, and appearance quality of soft rice in response to different nitrogen applications. The results showed that under the moderate nitrogen application (270 N), the soft rice exhibited the highest AGPase activity, the highest large‐starch granule content, and the lowest chalkiness. Under the highest nitrogen application (360 N), the soft rice exhibited the highest GBSS and DBE activity and the lowest SBE activity, the highest content of long‐branched amylopectin, the lowest relative crystallinity, the fewest ordered structures, the most amorphous structures, the largest semi‐crystalline lamellar thicknesses, and the highest transparency of chalk‐free rice. In conclusion, moderate nitrogen fertilization (270 N) improved the AGPase activity, which leaded to fuller starch granules and more compact endosperm in soft rice. Thus, the grain chalkiness of soft rice decreased. Continuous nitrogen application (0‐360 N) constantly increased the GBSS and DBE activity and reduced the SBE activity in soft rice, leading a lower content of short‐branched amylopectin and a higher content of long‐branched amylopectin in soft rice. Thus, the relative crystallinity and ordered structures of soft rice were reduced. These structures improved the transparency phenotype of soft rice.

Grain filling influences grain size and quality in cereal crops. The molecular mechanisms that regulate grain endosperm development remain elusive. In this study, we characterized a filling-defective and grain width mutant, fgw1, whose mutation increased rice seed width mainly via cell division and expansion in grains. Sucrose contents were higher but starch contents lower in the fgw1 mutant during the grain-filling stage, resulting in inferior endosperm of opaque, white appearance with loosely packed starch granules. Map-based cloning revealed that FGW1 encoded a protein containing DUF630/DUF632 domains, localized in the plasma membrane with preferential expression in the panicle. RNA interference in FGW1 resulted in increased grain width and weight, whereas overexpression of FGW1 led to slightly narrower kernels and better grain filling. In a yeast two-hybrid assay, FGW1 interacted directly with the 14–3–3 protein GF14f, bimolecular fluorescence complementation verified that the site of interaction was the membrane, and the mutated FGW1 protein failed to interact with GF14f. The expression of GF14f was down-regulated in fgw1, and the activities of AGPase, StSase, and SuSase in the endosperm of fgw1 increased similarly to those of a reported GF14f-RNAi. Transcriptome analysis indicated that FGW1 also regulates cellular processes and carbohydrate metabolism. Thus, FGW1 regulated grain formation via the GF14f pathway.

(1) The development of sweet potato storage roots is impacted by nitrogen (N) levels, with excessive nitrogen often impeding development. Starch synthesis enzymes such as sucrose synthase (SUS) and ADP-glucose pyrophosphorylase (AGPase) are pivotal in this context. Although the effects of excessive nitrogen on the formation of sweet potato storage roots are well documented, the specific responses of IbSUSs and IbAGPases have not been extensively reported on. (2) Pot experiments were conducted using the sweet potato cultivar “Pushu 32” at moderate (MN, 120 kg N ha−1) and excessive nitrogen levels (EN, 240 kg N ha−1). (3) Nine IbSUS and nine IbAGPase genes were categorized into three and two distinct subgroups based on phylogenetic analysis. Excessive nitrogen significantly (p < 0.05) suppressed the expression of IbAGPL1, IbAGPL2, IbAGPL4, IbAGPL5, IbAGPL6, IbAGPS1, and IbAGPS2 in fibrous roots and IbSUS2, IbSUS6, IbSUS7, IbSUS8, IbSUS9, IbAGPL2, and IbAGPL4 in storage roots, and then significantly (p < 0.05) decreased the SUS and AGPase activities and starch content of fibrous root and storage root, ultimately reducing the storage root formation of sweet potato. Excessive nitrogen extremely significantly (p < 0.01) enhanced the expression of IbAGPL3, which was strongly negatively correlated with the number and weight of storage roots per plant. (4) IbAGPL3 may be a key gene in the response to excessive nitrogen stress and modifying starch synthesis in sweet potato.

Grain size is one of the critical factors determining rice yield. Previous studies have found the grain-size-regulating function of Gα in rice. However, the regulatory mechanism underlying the development of rice grain mediated by Gα is still unclear. To reveal the functional mechanism of Gα in grain size regulation, a mutant of Gα (Gα-Cas9) was firstly constructed through a CRISPR/Cas9 strategy and was then grown in a greenhouse and field. The results showed that the seed length, plant height, 1000-grain weight, and spike length were significantly decreased in Gα-Cas9 compared to wild-type (WT) Pi-4b. During the grain filling stage, the increase in the grain dry weight of Pi-4b occurred earlier than that of Gα-Cas9. The total starch content and amylose content of matured grains of Pi-4b were higher than those of Gα-Cas9. Secondly, transcriptome sequencing analysis of Gα-Cas9 and Pi-4b during grain filling was performed to elucidate the functional pathways regulated by Gα. In total, 2867 and 4534 differentially expressed genes (DEGs) were discovered at 5 DAF and 10 DAF, and the starch and sucrose metabolism pathway enriched by DEGs was involved in grain size regulation mediated by Gα. Gα regulated the expression of starch-synthesis-related genes during grain filling, and the Gα protein interacted with OsNYC4 to trigger the sugar signaling pathway to promote starch accumulation in grain. Additionally, the GW2-WG1-OsbZIP47 pathway was switched off by Gα to relieve the inhibition of rice grain development. In this study, the results should provide new insights into the G protein signal transduction pathway.

Increasing grain weight under dense planting conditions can further improve maize yield. 6-BA is known to be involved in regulating grain development and influencing grain weight. Maize grain development is closely linked to starch accumulation and hormone levels. In this work, the effects of applying 6-BA at the flowering stage under high density on the grain filling characteristics, starch content, starch synthesis critical enzyme activity, and endogenous hormones levels of maize grains (including inferior grains (IGs) and superior grains (SGs)) of two high-yielding summer maize varieties widely cultivated in China were investigated. The findings indicated that applying 6-BA significantly improved maize yield compared to the control, mainly as a result of increased grain weight due to a faster grain filling rate. Additionally, the activities of enzymes associated with starch synthesis, including sucrose synthase (SuSy), ADP-glucose pyrophosphorylase (AGPase), granule-bound starch synthase (GBSS), soluble starch synthase (SSS), and starch branching enzyme (SBE), were all increased following 6-BA application, thus facilitating starch accumulation in the grains. Applying 6-BA also increased the zeatin riboside (ZR), indole-3-acetic acid (IAA), and abscisic acid (ABA) levels, and reduced the gibberellin (GA3) level in the grains, which further improved grain filling. It is worth noting that IG had a poorer filling process than SG, possibly due to the low activities of critical enzymes for starch synthesis and imbalanced endogenous hormones levels. However, IG responded more strongly to exogenous 6-BA than SG. It appears that applying 6-BA is beneficial in improving filling characteristics, promoting starch accumulation by enhancing the activities of critical enzymes for starch synthesis, and altering endogenous hormones levels in the grains, thus improving grain filling and increasing the final grain weight and yield of maize grown under crowded conditions. These results provide theoretical and technical support for the further utilization of exogenous hormones in high-density maize production. [ABSTRACT FROM AUTHOR]

Drip irrigation has a close relationship with the growth and development of maize grains and yield formation in semiarid areas. To explore the response mechanism of grain yield formation to drip irrigation quotas, a 2-year pond planting experiment was conducted under controlled conditions, by using two maize varieties with differences in drought resistance as experimental materials. Six treatments were set up, including CK1 (drought-resistant variety, 500 mm), T1 (drought-resistant variety, 350 mm), T2 (drought-resistant variety, 200 mm), CK2 (drought-sensitive variety, 500 mm), T3 (drought-sensitive variety, 350 mm), and T4 (drought-sensitive variety, 200 mm). The changes in maize grain filling characteristics, related hormones, enzyme activity related to starch synthesis, sugars and amino acids contents, and yield were analysed. The results showed that 100-grain weight at different filling times, filling rate, average filling rate, auxin, cytokinin, acid sucrose invertase, sucrose synthase, starch synthase, and adenosine diphosphate glucose pyro phosphorylase activities in maize grains decreased and the abscisic acid content and content of various amino acids and sugars in grains increased with the decrease in drip irrigation quota. The percentage of changes in drought-sensitive maize varieties was relatively high. The maize yield decreased with the decrease in drip irrigation quota. In summary, there was no significant difference in grain filling characteristics, hormone content, starch synthesis enzyme activity, and yield between maize treated with T1 (drought-resistant variety, 350 mm) and the control treatment. This effectively maintained grain growth and yield formation, achieving the goal of water saving and stable yields. [ABSTRACT FROM AUTHOR]

ADP-glucose pyrophosphorylase (AGPase) influences cereal productivity. There are few reports on the function of cytosolic AGPase small subunit in bread wheat (TaAGPS). In the present study, TaAGPS was preferentially expressed in developing endosperm during grain-filling stages in bread wheat. TaAGPS allelic variations were characterized in 143 wheat accessions by PacBio RS II sequencing. Two haplotypes (TaAGPS-7A-TG and TaAGPS-7A-CT) of TaAGPS-7A were identified and corresponding functional markers were developed, whereas no variants of TaAGPS-7B and TaAGPS-7D were detected. TaAGPS-7A was associated with thousand-kernel weight (TKW) by haplotype–trait association analysis in two populations. Near-isogenic lines (NILs) with TaAGPS-7A-TG showed higher TKW and total kernel starch content than those with TaAGPS-7A-CT, owing to the higher AGPase activity of TaAGPS-7A-TG than TaAGPS-7A-CT both in vitro and in vivo. Overexpression of TaAGPS-7A-TG in bread wheat doubled the transcription levels of TaAGPS and increased AGPase activity by 55.7%, resulting in a 3.0-g higher TKW than in the wild type (WT). Knockdown of TaAGPS led to reduced expression of TaAGPS, AGPase activity, and TKW than in the WT. Thus, owing to the 218th amino acid change of Ser to Ala in TaAGPS-7A, the favorable haplotype TaAGPS-7A-TG showed higher AGPase activity, resulting in higher kernel starch content and grain weight. This finding could be applied to increasing starch content and grain weight in bread wheat.

Soluble starch synthases (SSs) play important roles in the synthesis of cassava starch. However, the expression characteristics of the cassava SSs genes have not been elucidated. In this study, the MeSSIII-1 gene and its promoter, from SC8 cassava cultivars, were respectively isolated by PCR amplification. MeSSIII-1 protein was localized to the chloroplasts. qRT-PCR analysis revealed that the MeSSIII-1 gene was expressed in almost all tissues tested, and the expression in mature leaves was 18.9 times more than that in tuber roots. MeSSIII-1 expression was induced by methyljasmonate (MeJA), abscisic acid (ABA), and ethylene (ET) hormones in cassava. MeSSIII-1 expression patterns were further confirmed in proMeSSIII-1 transgenic cassava. The promoter deletion analysis showed that the −264 bp to −1 bp MeSSIII-1 promoter has basal activity. The range from −1228 bp to −987 bp and −488 bp to −264 bp significantly enhance promoter activity. The regions from −987 bp to −747 bp and −747 bp to −488 bp have repressive activity. These findings will provide an important reference for research on the potential function and transcriptional regulation mechanisms of the MeSSIII-1 gene and for further in-depth exploration of the regulatory network of its internal functional elements.

Although soybean and chickpea belong to the legume family, their seed starch content is very different. Currently, many studies focus on the molecular mechanisms of starch synthesis within a single species. However, the key genes and regulatory relationships responsible for the difference in seed starch content between the two species remain unknown. To elucidate the molecular mechanisms responsible for the above difference, multi-omics and bioinformatics analyses were used here to analyze gene expression patterns, protein–protein interaction networks, gene-transcription factor co-expression networks, and miRNA–gene regulatory relationships based on reported sucrose and starch metabolic genes in Arabidopsis. The results were as follows. First, seven differential expression genes of the two species in starch synthesis metabolism, including GBSS1, APL3, APS1, SS2, PTST, SBE2, and ISA, and the starch degradation gene BMY in soybean and chickpea, may contribute to their seed starch content differences. Then, the protein–protein interaction between DPEs and PHS may facilitate seed starch synthesis in chickpea. Finally, the positive regulation of two starch degradation genes (GmBMY and GmPHS) and four sucrose metabolism genes (GmHXK, GmPFK, GmTPS, and GmFRK) by transcription factors may lead to lower seed starch content in soybean. This study elucidates the possible molecular mechanisms underlying the difference in seed starch content between the two species and addresses the scientific problem of why soybean seeds have lower starch content than chickpea seeds.

In order to investigate the crucial role of ZmBT1 in starch accumulation during maize grain development and analyze the expression and distribution of ZmBT1 in various maize tissues, we prepared a polyclonal antibody. Specifically, we successfully expressed the recombinant plasmid pGEX-6p-ZmBT1-C (382-437aa) and purified Gst-ZmBT1-C as the antigen for antibody preparation. Our results confirmed that the ZmBT1 protein in maize tissues can be specifically recognized by the ZmBT1 antibody. Through Western blotting, we observed that the expression protein of ZmBT1 varied by tissues, with the highest content in the grain and endosperm. Furthermore, we employed a combination of Western blotting and quantitative real-time PCR to show that the expression level of ZmBT1 can be influenced by plant hormones. This finding suggests that ZmBT1 plays a critical role in the accumulation of starch and opens up new avenues for functional studies of this protein. [ABSTRACT FROM AUTHOR]

This study investigates carbon partitioning in the developing endosperm of a European variety of spring wheat subjected to moderately elevated daytime temperatures (27°C/16°C d/night) from anthesis to grain maturity. Elevated daytime temperatures caused significant reductions in both fresh and dry weights and reduced the starch content of harvested grains compared to plants grown under a 20°C/16°C d/night regimen. Accelerated grain development caused by elevated temperatures was accounted for by representing plant development as thermal time (°C DPA). We examined the effects of high-temperature stress (HTS) on the uptake and partitioning of [U-14C]-sucrose supplied to isolated endosperms. HTS caused reduced sucrose uptake into developing endosperms from the second major grain-filling stage (approximately 260°C DPA) up to maturity. Enzymes involved in sucrose metabolism were unaffected by HTS, whereas key enzyme activities involved in endosperm starch deposition such as ADP-glucose pyrophosphorylase and soluble isoforms of starch synthase were sensitive to HTS throughout grain development. HTS caused a decrease in other major carbon sinks such as evolved CO2, ethanol-soluble material, cell walls and protein. Despite reductions in the labeling of carbon pools caused by HTS, the relative proportions of sucrose taken up by endosperm cells allocated to each cellular pool remain unchanged, except for evolved CO2, which increased under HTS and may reflect enhanced respiratory activity. The results of this study show that moderate temperature increases can cause significant yield reductions in some temperate wheat cultivars chiefly through three effects: reduced sucrose uptake by the endosperm, reduced starch synthesis and increased partitioning of carbon into evolved CO2. [ABSTRACT FROM AUTHOR]

With the continuous change of global climate, the frequency of low-temperature stress (LTS) in spring increased greatly, which led to the increase of wheat yield decline. The effects of LTS at booting on grain starch synthesis and yield were examined in two wheat varieties with differing low-temperature sensitivities (insensitive variety Yannong 19 and sensitive variety Wanmai 52). A combination of potted and field planting was employed. For LTS treatment at booting, the wheat plants were placed in a climate chamber for 24 h at -2°C, 0°C or 2°C from 19:00 to 07:00 then 5°C from 07:00 to 19:00. They were then returned to the experimental field. The effects of flag leaf photosynthetic characteristics, the accumulation and distribution of photosynthetic products, enzyme activity related to starch synthesis and relative expression, the starch content, and grain yield were determined. LTS at booting caused a significant reduction in the net photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr) of the flag leaves at filling. The development of starch grains in the endosperm is also hindere, there are obvious equatorial grooves observed on the surface of the A-type starch granules, and a reduction in the number of Btype starch granules. The abundance of 13C in the flag leaves and grains decreased significantly. LTS also caused a significant reduction in translocation amount of pre-anthesis stored dry matte from vegetative organs to grains and amount of post-anthesis transfer of accumulated dry matte into grains, and the distribution rate of dry matter in the grains at maturity. The grain filling time was shortened, and the grain filling rate decreased. A decrease in the activity and relative expression of enzymes related to starch synthesis was also observed, with a decrease in the total starch content. As a result, a decrease in the grain number per panicle and 1000-grain weight were also observed. These findings highlight the underlying physiological cause of decreased starch content and grain weight after LTS in wheat. [ABSTRACT FROM AUTHOR]

Rice is a water intensive crop and soil water conditions affect rice yield and quality. However, there is limited research on the starch synthesis and accumulation of rice under different soil water conditions at different growth stages. Thus, a pot experiment was conducted to explore the effects of IR72 (indica) and Nanjing (NJ) 9108 (japonica) rice cultivars under flood-irrigated treatment (CK, 0 kPa), light water stress treatment (L, -20 ± 5 kPa), moderate water stress treatment (M, -40 ± 5 kPa) and severe water stress treatment (S, -60 ± 5 kPa) on the starch synthesis and accumulation and rice yield at booting stage (T1), flowering stage (T2) and filling stage (T3), respectively. Under LT treatment, the total soluble sugar and sucrose contents of both cultivars decreased while the amylose and total starch contents increased. Starch synthesis-related enzyme activities and their peak activities at mid-late growth stage increased as well. However, applying MT and ST treatments produced the opposite effects. The 1000-grain weight of both cultivars increased under LT treatment while the seed setting rate increased only under LT3 treatment. Compared with CK, water stress at booting stage decreased grain yield. The principal component analysis (PCA) showed that LT3 got the highest comprehensive score while ST1 got lowest for both cultivars. Furthermore, the comprehensive score of both cultivars under the same water stress treatment followed the trend of T3 > T2 > T1, and NJ 9108 had a better drought-resistant ability than IR72. Compared with CK, the grain yield under LT3 increased by 11.59% for IR72 and 16.01% for NJ 9108, respectively. Overall, these results suggested that light water stress at filling stage could be an effective method to enhance starch synthesis-related enzyme activities, promote starch synthesis and accumulation and increase grain yield. [ABSTRACT FROM AUTHOR]

In agricultural production, temperature and moisture are important factors affecting grain yield and quality. Although moderate drought at the grain-filling stage can effectively alleviate the damage caused by high temperature, the specific regulatory mechanism driving the effect of moderate drought at the high temperature on starch synthesis is still unclear. To explore the effects and mechanisms of high temperature and moderate drought on rice starch synthesis at the grain-filling stage, the activities of enzymes and expression levels of the genes involved in starch synthesis under four different treatments involving high temperature and/or water stress (CK, HT, WS, and HT+WS) were investigated in this study. The starch synthesis of a japonica inbred rice was measured under the four treatments during the grain filling. The results show that the effects of high temperature and moderate drought on grain filling mainly occur in the inferior grains of rice. Through the regulation of enzymes involved in starch synthesis and the expression levels of their main genes, the synthesis of rice starch can be affected. Therefore, the high temperature and moderate drought were antagonistic, and moderate drought can alleviate the damage to grain quality at a high temperature by improving the starch synthesis of inferior grains in japonica rice. This study provides a basis for stress-resistance cultivation and breeding strategies of rice with high temperature tolerance.

Introduction: Approximately 50% of irrigation water is saved during dripirrigation of rice, which has tremendous potential for water-saving agriculture, particularly in areas where water resources are scarce. However, the grain yield and quality of drip-irrigated rice are adversely affected. Methods: In this study, we investigated the effects of different irrigation strategies on the grain yield and quality of drip-irrigated rice using field experiments. Four irrigation treatments were studied: whole growing season flooding (FI), whole growing season normal drip irrigation (DI, soil relative moisture (RSM) was maintained in the range of 90-100%), pre-anthesis drip irrigation and postanthesis water stress (SAF, the RSM was maintained in the range of 80-90% after anthesis), pre-anthesis drip irrigation, and post-anthesis flooding (FAF). Results: The results showed that grain yield, harvest index, seed setting rate and 1000 grain weight in DI and SAF were significantly lower than in FI and FAF. These parameters were not significantly different between FI and FAF but were significantly greater in DI than in SAF. Compared with FI and FAF, the source capacity, source activity time, and sink activity of DI and SAF decreased, and the sink-source difference increased. The sink-source difference had a significant negative correlation with rice yield and 1000 grain weight. The activities of ADPglucose pyrophosphorylase, starch branching enzyme, and amylopectin content in grains in the middle panicles of FAF were significantly higher than those of DI and SAF. SAF resulted in increased amylose/amylopectin ratio and total protein content in grains but decreased proportion of glutenin in total protein. Irrigation after anthesis of drip-irrigated rice narrowed the difference between sink sources in rice plants, increased the grain yield and harvest index by 29.2% and 11%, respectively, compared to DI, increased water productivity by 19% compared to FI, and improved the grain quality of drip-irrigated rice. Discussion: This study highlights that post-anthesis sufficient irrigation of drip-irrigated rice plays a positive role in maintaining the source-sink balance. This study serves as a foundation for the development of more effective rice farming methods that conserve water, while increasing the grain yield and quality of drip-irrigated rice. [ABSTRACT FROM AUTHOR]

Sorghum is the fifth most important cereal crop. Here we performed molecular genetic analyses of the 'SUGARY FETERITA' (SUF) variety, which shows typical sugary endosperm traits (e.g., wrinkled seeds, accumulation of soluble sugars, and distorted starch). Positional mapping indicated that the corresponding gene was located on the long arm of chromosome 7. Within the candidate region of 3.4 Mb, a sorghum ortholog for maize Su1 (SbSu) encoding a starch debranching enzyme ISA1 was found. Sequencing analysis of SbSu in SUF uncovered nonsynonymous single nucleotide polymorphisms (SNPs) in the coding region, containing substitutions of highly conserved amino acids. Complementation of the rice sugary-1 (osisa1) mutant line with the SbSu gene recovered the sugary endosperm phenotype. Additionally, analyzing mutants obtained from an EMSinduced mutant panel revealed novel alleles with phenotypes showing less severe wrinkles and higher Brix scores. These results suggested that SbSu was the corresponding gene for the sugary endosperm. Expression profiles of starch synthesis genes during the grain-filling stage demonstrated that a loss-offunction of SbSu affects the expression of most starch synthesis genes and revealed the fine-tuned gene regulation in the starch synthetic pathway in sorghum. Haplotype analysis using 187 diverse accessions from a sorghum panel revealed the haplotype of SUF showing severe phenotype had not been used among the landraces and modern varieties. Thus, weak alleles (showing sweet and less severe wrinkles), such as in the abovementioned EMS-induced mutants, are more valuable for grain sorghum breeding. Our study suggests that more moderate alleles (e.g. produced by genome editing) should be beneficial for improving grain sorghum. [ABSTRACT FROM AUTHOR]

During grain filling, starch and other nutrients accumulate in the endosperm; this directly determines grain yield and grain quality in crops such as rice (Oryza sativa), maize (Zea mays), and wheat (Triticum aestivum). Grain filling is a complex trait affected by both intrinsic and environmental factors, making it difficult to explore the underlying genetics, molecular regulation, and the application of these genes for breeding. With the development of powerful genetic and molecular techniques, much has been learned about the genes and molecular networks related to grain filling over the past decades. In this review, we highlight the key factors affecting grain filling, including both biological and abiotic factors. We then summarize the key genes controlling grain filling and their roles in this event, including regulators of sugar translocation and starch biosynthesis, phytohormone‐related regulators, and other factors. Finally, we discuss how the current knowledge of valuable grain filling genes could be integrated with strategies for breeding cereal varieties with improved grain yield and quality. [ABSTRACT FROM AUTHOR]

The kernel serves as a storage organ for various nutrients and determines the yield and quality of maize. Understanding the mechanisms regulating kernel development is important for maize production. In this study, a small-kernel mutant smk7a of maize was characterized. Cytological observation suggested that the development of the endosperm and embryo was arrested in smk7a in the early development stage. Biochemical tests revealed that the starch, zein protein, and indole-3-acetic acid (IAA) contents were significantly lower in smk7a compared with wild-type (WT). Consistent with the defective development phenotype, transcriptome analysis of the kernels 12 and 20 days after pollination (DAP) revealed that the starch, zein, and auxin biosynthesis-related genes were dramatically downregulated in smk7a. Genetic mapping indicated that the mutant was controlled by a recessive gene located on chromosome 2. Our results suggest that disrupted nutrition accumulation and auxin synthesis cause the defective endosperm and embryo development of smk7a. [ABSTRACT FROM AUTHOR]

Increasing grain weight under dense planting conditions can further improve maize yield. 6-BA is known to be involved in regulating grain development and influencing grain weight. Maize grain development is closely linked to starch accumulation and hormone levels. In this work, the effects of applying 6-BA at the flowering stage under high density on the grain filling characteristics, starch content, starch synthesis critical enzyme activity, and endogenous hormones levels of maize grains (including inferior grains (IGs) and superior grains (SGs)) of two high-yielding summer maize varieties widely cultivated in China were investigated. The findings indicated that applying 6-BA significantly improved maize yield compared to the control, mainly as a result of increased grain weight due to a faster grain filling rate. Additionally, the activities of enzymes associated with starch synthesis, including sucrose synthase (SuSy), ADP-glucose pyrophosphorylase (AGPase), granule-bound starch synthase (GBSS), soluble starch synthase (SSS), and starch branching enzyme (SBE), were all increased following 6-BA application, thus facilitating starch accumulation in the grains. Applying 6-BA also increased the zeatin riboside (ZR), indole-3-acetic acid (IAA), and abscisic acid (ABA) levels, and reduced the gibberellin (GA3) level in the grains, which further improved grain filling. It is worth noting that IG had a poorer filling process than SG, possibly due to the low activities of critical enzymes for starch synthesis and imbalanced endogenous hormones levels. However, IG responded more strongly to exogenous 6-BA than SG. It appears that applying 6-BA is beneficial in improving filling characteristics, promoting starch accumulation by enhancing the activities of critical enzymes for starch synthesis, and altering endogenous hormones levels in the grains, thus improving grain filling and increasing the final grain weight and yield of maize grown under crowded conditions. These results provide theoretical and technical support for the further utilization of exogenous hormones in high-density maize production.

Drip irrigation has a close relationship with the growth and development of maize grains and yield formation in semiarid areas. To explore the response mechanism of grain yield formation to drip irrigation quotas, a 2-year pond planting experiment was conducted under controlled conditions, by using two maize varieties with differences in drought resistance as experimental materials. Six treatments were set up, including CK1 (drought-resistant variety, 500 mm), T1 (drought-resistant variety, 350 mm), T2 (drought-resistant variety, 200 mm), CK2 (drought-sensitive variety, 500 mm), T3 (drought-sensitive variety, 350 mm), and T4 (drought-sensitive variety, 200 mm). The changes in maize grain filling characteristics, related hormones, enzyme activity related to starch synthesis, sugars and amino acids contents, and yield were analysed. The results showed that 100-grain weight at different filling times, filling rate, average filling rate, auxin, cytokinin, acid sucrose invertase, sucrose synthase, starch synthase, and adenosine diphosphate glucose pyro phosphorylase activities in maize grains decreased and the abscisic acid content and content of various amino acids and sugars in grains increased with the decrease in drip irrigation quota. The percentage of changes in drought-sensitive maize varieties was relatively high. The maize yield decreased with the decrease in drip irrigation quota. In summary, there was no significant difference in grain filling characteristics, hormone content, starch synthesis enzyme activity, and yield between maize treated with T1 (drought-resistant variety, 350 mm) and the control treatment. This effectively maintained grain growth and yield formation, achieving the goal of water saving and stable yields.

Drought stress at jointing–booting directly affects plant growth and productivity in rice. Limited by natural factors, the jointing and booting stages of short-growth-period rice varieties are highly overlapped in high-latitude areas, which are more sensitive to water deficit. However, little is known about the dry matter translocation in rice and the strategies of starch synthesis and filling of superior and inferior grains under different drought stress was unclear. In this study, the rice plants were subjected to three degrees of drought stress (−10 kPa, −25 kPa, −40 kPa) for 15 days during the jointing–booting stage; we investigated dry matter accumulation and translocation, grain filling and enzyme activities to starch synthesis of superior and inferior grains in rice with overlapping growth stages from 2016 to 2017. The results showed that drought stress significantly reduced dry matter accumulation in the stems and leaves. Mild and moderate drought increased dry matter translocation efficiency. However, severe drought stress largely limited the dry matter accumulation and translocation. A large amount of dry matter remains in vegetative organs under severe drought stress. The high content in NSC in stem and sheath plays a key role in resisting drought stress. The drought stress at jointing–booting directly caused a change in the grain filling strategy. Under moderate and severe drought, the grain-filling active period of the superior grains was shortened to complete the necessary reproductive growth. The grain-filling active period of the inferior grains was significantly prolonged to avoid a decrease in grain yield. The significant decrease in the grain-filling rate of the superior and inferior grains caused a reduction in the thousand-grain weight. In particular, the influence of the grain-filling rate of inferior grains on the thousand-grain weight was more significant. Drought stress changed the starch synthesis strategies of the superior and inferior grains. Soluble starch synthase and starch branching enzyme activities of inferior grains increased significantly under drought stress. GBSS activity was not sensitive to drought stress. Therefore, amylose content was decreased and amylopectin synthesis was enhanced under drought stress, especially in inferior grains. [ABSTRACT FROM AUTHOR]

Low light stress increases the chalkiness of rice; however, this effect has not been fully characterized. In this study, we demonstrated that low light resulted in markedly decreased activity of ADP-glucose pyrophosphorylase in the grains and those of sucrose synthase and soluble starch synthase in the early period of grain filling. Furthermore, low light also resulted in decreased activities of granule-bound starch synthase and starch branching enzyme in the late period of grain filling. Therefore, the maximum and mean grain filling rates were reduced but the time to reach the maximum grain filling rates and effective grain filling period were increased by low light. Thus, it significantly decreased the grain weight at the maximum grain filling rate and grain weight and retarded the endosperm growth and development, leading to a loose arrangement of the amyloplasts and an increase in the chalkiness of the rice grains. Compared to the grains at the top panicle part, low light led to a greater decrease in the grain weight at the maximum grain filling rate and time to reach the grain weight at the maximum grain filling rate at the bottom panicle part, which contributed to an increase in chalkiness by increasing the rates of different chalky types at the bottom panicle part. In conclusion, low light disturbed starch synthesis in grains, thereby impeding the grain filling progress and increasing chalkiness, particularly for grains at the bottom panicle part. [ABSTRACT FROM AUTHOR]

The physiological implications of autophagy in plant cells have not been fully elucidated. Therefore, we investigated the consequences of autophagy in the moss Physcomitrella by measuring biochemical parameters (fresh and dry weights; starch, amino acid, carbohydrate, and NH3 content) in wild-type (WT) and autophagy-deficient atg5 Physcomitrella cells. We found higher starch levels and a higher net starch synthesis rate in WT cells than in atg5 cells cultured in a glucose-containing culture medium, whereas net starch degradation was similar in the two strains cultured in a glucose-deficient culture medium. Additionally, the treatment of cells with the autophagy inhibitor 3-methyladenine suppressed starch synthesis. Loading bovine serum albumin into atg5 cells through endocytosis, i.e., supplying proteins to vacuoles in the same way as through autophagy, accelerated starch synthesis, whereas loading glutamine through the plasma membrane had no such effect, suggesting that Physcomitrella cells distinguish between different amino acid supply pathways. After net starch synthesis, NH3 levels increased in WT cells, although the change in total amino acid content did not differ between WT and atg5 cells, indicating that autophagy-produced amino acids are oxidized rapidly. We conclude that autophagy promotes starch synthesis in Physcomitrella by supplying the energy obtained by oxidizing autophagy-produced amino acids. [ABSTRACT FROM AUTHOR]

Iron (Fe) storage in cereal seeds is the principal source of dietary Fe for humans. In maize (Zea mays), the accumulation of Fe in seeds is known to be negatively correlated with crop yield. Hence, it is essential to understand the underlying mechanism, which is crucial for developing and breeding maize cultivars with high yields and high Fe concentrations in the kernels. Here, through the successful application of in vitro kernel culture, we demonstrated that excess Fe supply in the medium caused the kernel to become collapsed and lighter in color, consistent with those found in yellow strip like 2 (ysl2, a small kernel mutant), implicated a crucial role of Fe concentration in kernel development. Indeed, over-accumulation of Fe in endosperm inhibited the abundance and activity of ADP-glucose pyrophosphorylase (AGPase) and the kernel development defect was alleviated by overexpression of Briittle 2 (Bt2, encoding a small subunit of AGPase) in ysl2 mutant. Imaging and quantitative analyses of reactive oxygen species (ROS) and cell death showed that Fe stress-induced ROS burst and severe DNA damage in endosperm cells. In addition, we have successfully identified candidate genes that are associated with iron homeostasis within the kernel, as well as upstream transcription factors that regulate ZmYSL2 by yeast one-hybrid screening. Collectively, our study will provide insights into the molecular mechanism of Fe accumulation-regulated seed development and promote the future efficient application of Fe element in corn improvement., (© 2024 Wiley Periodicals LLC.)

Abstract Background Chestnut seeds are important kinds of edible nuts rich in starch and protein. The characteristics and nutrient contents of chestnut have been found to show obvious metaxenia effects in previous studies. To improve the understanding of the effect of metaxenia on chestnut starch and sucrose metabolism, this study used three varieties of chestnut, ‘Yongfeng 1’, ‘YongRen Zao’ and ‘Yimen 1’, as male parents to pollinate the female parent, ‘Yongfeng 1’, and investigated the mechanisms of starch and sucrose metabolism in three starch accumulation stages (70 (S1), 82 (S2), and 94 (S3) days after pollination, DAP) in chestnut seed kernels. Result Most carbohydrate metabolism genes were highly expressed in YFF (self-pollinated ‘Yongfeng 1’) in stage S2 and in YFR (‘Yongfeng 1’ × ‘Yongren Zao’) and YFM (‘Yongfeng 1’ × ‘Yimen 1’) in stage S3. In stage S3, hub genes encoding HSF_DNA-binding, ACT, Pkinase, and LIM proteins and four transcription factors were highly expressed, with YFF showing the highest expression, followed by YFR and YFM. In addition, transcriptome analysis of the kernels at 70, 82 and 94 DAP showed that the starch granule-bound starch synthase (EC 2.4.1.242) and ADP-glucose pyrophosphorylase (EC 2.7 .7.27) genes were actively expressed at 94 DAF. Chestnut seeds regulate the accumulation of soluble sugars, reducing sugars and starch by controlling glycosyl transferase and hydrolysis activity during development. Conclusion These results and resources have important guiding significance for further research on starch and sucrose metabolism and other types of metabolism related to chestnut metaxenia.

In order to investigate the crucial role of ZmBT1 in starch accumulation during maize grain development and analyze the expression and distribution of ZmBT1 in various maize tissues, we prepared a polyclonal antibody. Specifically, we successfully expressed the recombinant plasmid pGEX-6p-ZmBT1-C (382-437aa) and purified Gst-ZmBT1-C as the antigen for antibody preparation. Our results confirmed that the ZmBT1 protein in maize tissues can be specifically recognized by the ZmBT1 antibody. Through Western blotting, we observed that the expression protein of ZmBT1 varied by tissues, with the highest content in the grain and endosperm. Furthermore, we employed a combination of Western blotting and quantitative real-time PCR to show that the expression level of ZmBT1 can be influenced by plant hormones. This finding suggests that ZmBT1 plays a critical role in the accumulation of starch and opens up new avenues for functional studies of this protein.

Canna edulis Ker-Gawl and Canna indica L. are species belonging to the Cannaceae family and both have a very high economic value. Here, we aimed to assemble genomes of C. edulis and C. indica at the chromosome level to generate a reference genome for the Cannaceae family. We also comparatively analyzed the genomes of C. edulis and C. indica and examined the molecular mechanisms responsible for the remarkable differences in plant characteristics in C. edulis varieties. Our results indicated that genomewide duplication events had recently occurred in C. edulis and C. indica. The comparative analysis of the genomes of C. edulis and C. indica revealed that C. edulis exhibited a remarkable level of replication of genes in the starch and sucrose metabolic pathways, especially during sucrose hydrolysis. This finding is consistent with the fact that the starch content of the C. edulis tuber is higher than that of C. indica. Simplified genome re-sequencing revealed the population structure of 241 C. edulis genes, and a genome-wide association study of leaf traits revealed the location of key genes related to leaf color and morphology. These findings extend our understanding of Cannaceade at the molecular level, and provide an effective theoretical basis for further study and utilization of Cannaceae plants. [ABSTRACT FROM AUTHOR]

Starch directly determines the grain yield and quality. The key enzymes participating in the process of starch synthesis have been cloned and characterized. Nevertheless, the regulatory mechanisms of starch synthesis remain unclear. In this study, we identified a novel starch regulatory gene, ZmCBM48-1, which contained a carbohydrate-binding module 48 (CBM48) domain. ZmCBM48-1 was highly expressed in the maize endosperm and was localized in the plastids. Compared with the wild type lines, the overexpression of ZmCBM48-1 in rice altered the grain size and 1000-grain weight, increased the starch content, and decreased the soluble sugar content. Additionally, the transgenic rice seeds exhibited an alterant endosperm cell shape and starch structure. Meanwhile, the physicochemical characteristics (gelatinization properties) of starch were influenced in the transgenic lines of the endosperm compared with the wild type seeds. Furthermore, ZmCBM48-1 played a positive regulatory role in the starch synthesis pathway by up-regulating several starch synthesis-related genes. Collectively, the results presented here suggest that ZmCBM48-1 acts as a key regulatory factor in starch synthesis, and could be helpful for devising strategies for modulating starch production for a high yield and good quality in maize endosperm. [ABSTRACT FROM AUTHOR]

Starch biosynthesis is a complex and highly controlled process that requires coordinated activities among multiple enzymes. Starch synthase III (SSIII) is the largest protein in the starch synthase complex and its function is to lengthen long-chain amylopectin in starch synthesis. It potentially affects the activity of other key enzymes in starch synthesis through protein–protein interactions; therefore, its function and regulation play a predominant role in starch synthesis. In this review, we summarized the main research of SSIII including its biochemical characteristics, structural features, expression atlas, and regulation means. Structural features and expressional analysis indicated that SSIIIa is the main functional protein in maize endosperm rather than SSIIIb-a and SSIIIb-b, even though they are similar in the tertiary structures. The regulation investigation of SSIIIa showed that there are 13 transcription factors that control the transcription of SSIIIa. Interaction network analysis showed that SSIIIa could be involved with ten other key enzymes in starch synthesis. In conclusion, this review considerably extends our understanding of SSIII and provides the theoretical basis for improving starch synthesis by SSIII in maize. [ABSTRACT FROM AUTHOR]

Improving grain filling in the presernt farming systems is crucial where grain filling is a concern due to the extreme use of chemical fertilizers (CF). A field experiment was conducted at the experimental station of Guangxi University, China in 2019 to test the hypothesis that cattle manure (CM) and poultry manure (PM) combined with CF could improve rice grain filling rate, yield, biochemical and qualitative attributes. A total of six treatments, i.e., no fertilizer (T1), 100% CF (T2), 60% CM + 40% CF (T3), 30% CM + 70% CF (T4), 60% PM + 40% CF (T5), and 30% PM + 70% CF (T6) were used in this study. Results showed that the combined treatment T6increased starch metabolizing enzymes activity (SMEs), such as ADP-glucose phosphorylase (ADGPase) by 8 and 12%, soluble starch synthase (SSS) by 7 and 10%, granule bound starch synthesis (GBSS) by 7 and 9%, and starch branching enzyme (SBE) by 14 and 21% in the early and late seasons, respectively, compared with T2. Similarly, higher rice grain yield, grain filling rate, starch, and amylose content were also recorded in combined treatments. In terms of seasons, higher activity of SMEs , grain starch, and amylose content was noted in the late-season compared to the early season. The increment in these traits was mainly attributed to a lower temperature in the late season during the grain filling period. Furthermore, our results suggested that an increment in starch accumulation and grain filling rate were mainly associated with the enhanced sink capacity by regulating key enzyme activities involved in Suc-to-starch conversion. In-addition, RT-qPCR analysis showed higher expression levels of AGPS2b, SSS1, GBSS1, and GBSE11b genes, which resultantly increased the activities of SMEs during the grain filling period under combined treatments. Linear regression analysis revealed that the activity of ADGPase, SSS, GBSS, and SBE were highly positively correlated with starch and amylose accumulation. Thus, we concluded that a combination of 30% N from PM or CM with 70% N from CF is a promising option in terms of improving rice grain yield and quality. Our study provides a sustainable fertilizer management strategy to enhance rice grain yield and quality at the lowest environmental cost.

Maize (Zea mays L.) is one of the most important food crops in the world, and starch is the main component of its endosperm. Transcriptional regulation plays a vital role in starch biosynthesis. However, it is not well understood in maize. We report the identification of the transcription factor ZmNAC126 and its role in regulation of starch synthesis in maize. Transcriptional expression of ZmNAC126 was higher in maize endosperm and kernels than in roots or stems. ZmNAC126 shared a similar expression pattern with starch synthesis genes during seed development, and its expression pattern was also consistent with the accumulation of starch. ZmNAC126 is a typical transcription factor with a transactivation domain between positions 201 and 227 of the amino acid sequence, is located in the nucleus, and binds to CACG repeats in vitro. Yeast one-hybrid assay revealed that ZmNAC126 bound the promoters of ZmGBSSI, ZmSSIIa, ZmSSIV, ZmISA1, and ZmISA2. Transient overexpression of ZmNAC126 in maize endosperm increased the activities of promoters pZmSh2, pZmBt2, pZmGBSSI, pZmSSIIIa, and pZmBT1 but inhibited the activities of pZmISA1 and pZmISA2. ZmNAC126 thus acts in starch synthesis by transcriptionally regulating targeted starch synthesis-related genes in maize kernels.

Now, lodging is a major constraint factor contributing to yield loss of maize (Zea mays L.) under high planting density. Chemical regulation and nitrogen fertilizer could effectively coordinate the relationship between stem lodging and maize yield, which significantly reduce lodging and improve the grain yield. The purpose of this study was to explore the effects of chemical regulation and different nitrogen application rates on lodging characteristics, grain filling and yield of maize under high density. For this, we established a field study during 2017 and 2018 growing seasons, with three nitrogen levels of N100 (100 kg ha–1), N200 (200 kg ha–1) and N300 (300 kg ha–1) at high planting density (90 000 plants ha–1), and applied plant growth regulator (Yuhuangjin, the mixture of 3% DTA-6 and 27% ethephon) at the 7th leaf. The results showed that chemical control increased the activities of phenylalanine ammonia-lyase (PAL), tyrosine ammonia-lyase (TAL), 4-coumarate:CoA ligase (4CL), and cinnamyl alcohol dehydrogenase (CAD), and increased the lignin, cellulose and hemicellulose contents at the bottom of the 3rd internode, which significantly reduced the lodging percentage. The lignin-related enzyme activities, lignin, cellulose and hemicellulose contents decreased with the increase of nitrogen fertilizer, which significantly increased the lodging percentage. The 200 kg ha–1 nitrogen application and chemical control increased the number, diameter, angle, volume, and dry weight of brace roots. The 200 kg ha–1 nitrogen application and chemical control significantly increased the activities of ADP-glucose pyrophosphorylase (AGPase), soluble starch synthase (SSS) and starch branching enzyme (SBE), which promoted the starch accumulation in grains. Additional, improved the maximum grain filling rate (Vmax) and mean grain filling rate (Vm), which promoted the grain filling process, significantly increased grain weight and grain number per ear, thus increased the final yield.

Summary: Starch synthesis makes a dramatic contribution to the yield and nutritional value of cereal crops. Although several starch synthesis enzymes and related regulators have been reported, the underlying regulatory mechanisms of starch synthesis remain largely unknown.OsMADS14 is a FRUITFULL (FUL)‐like MADS‐box gene in rice (Oryza sativa). Here we show that two null mutations of OsMADS14 result in a shrunken and chalky grain phenotype. It is caused by obviously defective compound starch granules and a significantly reduced content of both total starch and amylose in the endosperm.Transcriptomic profiling analyses revealed that the loss‐of‐function of OsMADS14 leads to significantly downregulated expression of many core starch synthesis genes, including OsAGPL2 and Waxy. Both in vitro and in vivo assays demonstrate that the OsMADS14 protein directly binds to stretches of DNA with a CArG‐box consensus in the putative regulatory regions of OsAGPL2 and Waxy. Protein–protein interaction experiments also suggest that OsMADS14 interacts with nuclear factor NF‐YB1 to promote the transcription of OsAGPL2 and Waxy.Our study thus demonstrates that OsMADS14 plays an essential role in the synthesis of storage starch and provides novel insights into the underlying molecular mechanism that may be used to improve rice cultivars by molecular breeding. [ABSTRACT FROM AUTHOR]

Noodles are an important food in Asia. Wheat starch is the most important component in Chinese noodles. Loss of the waxy genes leads to lower activity of starch synthesis enzymes and decreased amylose content that further affects starch properties and noodle quality. To study the effects of different waxy (Wx) protein subunits on starch biosynthesis and processing quality, the high-yielding wheat cultivar Jimai 22 was treated with the mutagen ethyl methane sulfonate (EMS) to produce a population of Wx lines and chosen 7 Wx protein combinations. The amylose content increased but swelling power decreased as the number of Wx proteins increased. Both GBSS activity and gene expression were the lowest for the waxy mutant, followed by the mutants with 1 Wx protein. The combinations of these mutant alleles lead to reductions in both RNA expression and protein levels. Noodles made from materials with 2 Wx protein subunits had the highest score, which agreed with peak viscosity. The influence of the Wx-B1 protein on amylose synthesis and noodle quality was the highest, whereas the influence of Wx-A1 protein was the lowest. Mutants with lower amylose content caused by the absence of 1 subunit, especially the Wx-B1 subunit, had superior noodle quality. Additionally, the identified mutant lines can be used as intermediate materials to improve wheat quality. [ABSTRACT FROM AUTHOR]

Summary: Starch accounts for over 80% of the total dry weight in cereal endosperm and determines the kernel texture and nutritional quality. Amyloplasts, terminally differentiated plastids, are responsible for starch biosynthesis and storage. We screened a series of rice mutants with floury endosperm to clarify the mechanism underlying amyloplast development and starch synthesis. We identified the floury endosperm19 (flo19) mutant which shows opaque of the interior endosperm. Abnormal compound starch grains (SGs) were present in the endosperm cells of the mutant. Molecular cloning revealed that the FLO19 allele encodes a plastid‐localized pyruvate dehydrogenase complex E1 component subunit α1 (ptPDC‐E1‐α1) that is expressed in all rice tissues. In vivo enzyme assays demonstrated that the flo19 mutant showed decreased activity of the plastidic pyruvate dehydrogenase complex. In addition, the amounts of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) were much lower in the developing flo19 mutant endosperm, suggesting that FLO19 participates in fatty acid supply for galactolipid biosynthesis in amyloplasts. FLO19 overexpression significantly increased seed size and weight, but did not affect other important agronomic traits, such as panicle length, tiller number and seed setting rate. An analysis of single nucleotide polymorphism data from a panel of rice accessions identified that the pFLO19L haplotype was positively associated with grain length, implying a potential application in rice breeding. In summary, our study demonstrates that FLO19 is involved in galactolipid biosynthesis which is essential for amyloplast development and starch biosynthesis in rice. [ABSTRACT FROM AUTHOR]

The sago palm (Metroxylon sagu Rottboll) is a tropical halophytic starch-producing, economically important crop palm mainly located in Southeast Asian countries. Recently, a genome survey was conducted on this palm using the Illumina sequencing platform, with a very low (21.5%) BUSCO genome completeness score, and most of them (∼78%) are either fragmented or missing. Thus, in this study, the sago palm genome completeness was further improved with the utilization of the Nanopore sequencing platform that produced longer reads. A hybrid genome assembly was conducted, and the outcome was a much complete sago palm genome with BUSCO completeness achieved at as high as 97.9%, with only ∼2% of them either fragmented or missing. The estimated genome size of the sago palm is 509,812,790 bp in this study. A sum of 33,242 protein-coding genes was revealed from the sago palm genome and around 96.39% of them had been functionally annotated. An investigation on the carbohydrate metabolism KEGG pathways also unearthed that starch synthesis was one of the major sago palm activities. The genome data obtained from this work is indispensable for future molecular evolutionary and genome-wide association studies on the economically important sago palm.