Starch Biosynthesis Research Articles

Potential Roles of the GRF Transcription Factors in Sorghum Internodes during Post-Reproductive Stages.

Growth-regulating factor (GRF) is a plant-specific family of transcription factors crucial for meristem development and plant growth. Sorghum (Sorghum bicolor L. Moench) is a cereal species widely used for food, feed and fuel. While sorghum stems are important biomass components, the regulation of stem development and the carbohydrate composition of the stem tissues remain largely unknown. Here, we identified 11 SbGRF-encoding genes and found the SbGRF expansion driven by whole-genome duplication events. By comparative analyses of GRFs between rice and sorghum, we demonstrated the divergence of whole-genome duplication (WGD)-derived OsGRFs and SbGRFs. A comparison of SbGRFs' expression profiles supports that the WGD-duplicated OsGRFs and SbGRFs experienced distinct evolutionary trajectories, possibly leading to diverged functions. RNA-seq analysis of the internode tissues identified several SbGRFs involved in internode elongation, maturation and cell wall metabolism. We constructed co-expression networks with the RNA-seq data of sorghum internodes. Network analysis discovered that SbGRF1, 5 and 7 could be involved in the down-regulation of the biosynthesis of cell wall components, while SbGRF4, 6, 8 and 9 could be associated with the regulation of cell wall loosening, reassembly and/or starch biosynthesis. In summary, our genome-wide analysis of SbGRFs reveals the distinct evolutionary trajectories of WGD-derived SbGRF pairs. Importantly, expression analyses highlight previously unknown functions of several SbGRFs in internode elongation, maturation and the potential involvement in the metabolism of the cell wall and starch during post-anthesis stages.

Source-sink synergy is the key unlocking sweet potato starch yield potential

Sweet potato starch is in high demand globally for food and industry. However, starch content is negatively correlated with fresh yield. It is urgent to uncover the genetic basis and molecular mechanisms underlying the starch yield of sweet potato. Here we systematically explore source-sink synergy-mediated sweet potato starch yield formation: the production, loading, and transport of photosynthates in leaves, as well as their unloading and allocation in storage roots, lead to starch content divergence between sweet potato varieties. Moreover, we find that six haplotypes of IbPMA1 encoding a plasma membrane H+-ATPase are significantly linked with starch accumulation. Overexpression of IbPMA1 in sweet potato results in significantly increased starch and sucrose contents, while its knockdown exhibits an opposing effect. Furthermore, a basic helix-loop-helix (bHLH) transcription factor IbbHLH49 directly targets IbPMA1 and activates its transcription. Overexpression of IbbHLH49 notably improves source-sink synergy-mediated fresh yield and starch accumulation in sweet potato. Both IbbHLH49 and IbPMA1 substantially influence sugar transport and starch biosynthesis in source and sink tissues. These findings expand our understanding of starch yield formation and provide strategies and candidate genes for high starch breeding in root and tuber crops.

Perspectives on Starch Structure, Function, and Synthesis in Relation to the Backbone Model of Amylopectin.

Understanding functionality of polysaccharides such as starch requires molecular representations that account for their functional characteristics, such as those related to gelatinization, gelation, and crystallization. Starch macromolecules are inherently very complex, and precise structures can only be deduced from large data sets to generate relational models. For amylopectin, the major, well-organized, branched part of starch, two main molecular representations describe its structure: the classical cluster model and the more recent backbone model. Continuously emerging data call for inspection of these models, necessary revisions, and adoption of the preferred representation. The accumulated molecular and functional data support the backbone model and it well accommodates our present knowledge related to the biosynthesis of starch. This Perspective focuses on our current knowledge of starch structure and functionality directly in relation to the backbone model of amylopectin.

Genome-Wide Identification, Bioinformatic Characterization, and Expression Profiling of Starch Synthase (SS) Genes in Foxtail Millet under Drought Condition

Millet, a vital and nutritionally dense cereal extensively cultivated in Sub-Saharan Africa, plays a key role in ensuring food security. This study investigates the starch synthase (SS) gene family, which is crucial for starch biosynthesis and influences various plant functions and stress responses. While the specific roles of SS genes in millet under drought conditions are not fully elucidated, this research provides a thorough analysis of the SS gene family in millet. A total of twelve millet SS genes (SiSSs) were identified and classified into four subfamilies (I–IV) through gene structure and phylogenetic analysis. The SiSS genes were unevenly distributed across millet chromosomes, with cis-acting elements associated with plant growth and stress defense being identified. Quantitative PCR (qPCR) revealed dynamic and varied expression patterns of SiSSs in different tissues under drought stress. Millet plants subjected to drought conditions showed higher tissue starch content and increased starch synthase activity compared to controls. Importantly, the expression levels of the twelve SiSSs were positively correlated with both starch content and synthase activity, suggesting their significant role in drought tolerance. This study enhances our understanding of the millet SS gene family and highlights the potential of these genes in breeding programs aimed at developing drought-resistant millet varieties. Further research is recommended to validate these findings and delve deeper into the mechanisms underlying drought tolerance.

Protein kinase MeSnRK2.3 positively regulates starch biosynthesis by interacting with the transcription factor MebHLH68 in cassava.

Starch biosynthesis involves numerous enzymes and is a crucial metabolic activity in plant storage organs. Sucrose non-fermenting related protein kinase 2 (SnRK2) is an abscisic acid (ABA)-dependent kinase and a significant regulatory enzyme in the ABA signaling pathway. However, whether SnRK2 kinases regulate starch biosynthesis is unclear. In this study, we identified that MeSnRK2.3, an ABA-dependent kinase, was highly expressed in the storage roots of cassava and was induced by ABA. Overexpression of MeSnRK2.3 in cassava significantly increased the starch content in the storage roots and promoted plant growth. MeSnRK2.3 was further found to interact with the cassava basic helix-loop-helix 68 (MebHLH68) transcription factor in vivo and in vitro. MebHLH68 directly bound to the promoters of sucrose synthase 1 (MeSUS1), granule-bound starch synthase I a (MeGBSSIa), and starch-branching enzyme 2.4 (MeSBE2.4), thereby upregulating their transcriptional activities. Additionally, MebHLH68 negatively regulated the transcriptional activity of sucrose phosphate synthase B (MeSPSB). Moreover, phosphorylated MebHLH68 by MeSnRK2.3 up-regulated the transcription activity of MeSBE2.4. These findings demonstrated that the MeSnRK2.3-MebHLH68 module connects the ABA signaling pathway and starch biosynthesis in cassava, thereby providing direct evidence of ABA-mediated participation in the sucrose metabolism and starch biosynthesis pathway.

Effects of high temperature on grain quality and enzyme activity in heat-sensitive versus heat-tolerant rice cultivars.

High-temperature (HT) stress significantly affects the quality of rice (Oryza sativa L.), although the underlying the mechanism remains unknown. Therefore, in the present study, we assessed protein components, amino acids, mineral element levels, starch biosynthesis enzyme activity and gene expression of two heat-sensitive and two heat-tolerant genotypes under HT treatment during early (from 1 to 10 days, T1) and mid-filling (from 11 to 20 days, T2) after anthesis. Except for one cultivar, most rice varieties exhibited increased levels of amylose, chalky degree and protein content, along with elevated cracked grains and pasting temperatures and, consequently, suppressed amino acid levels under HT stress. HT treatment also increased protein components, macro- (Mg, K, P and S) and microelements (Cu, Zn, and Mo) in the rice flour. Both HT treatments reduced the activity of ADP-glucose pyrophosphate, ground-bound starch synthase, as well as the relative ratio of amylose to total starch, at the same time increasing starch branch enzyme activity. The expression levels of OsAGPL2, OsSSS1 and OsSBE1 in all varieties exhibited the same trends as enzyme activity under HT treatment. High temperatures negatively affected rice quality during grain filling, which is related to heat tolerance and grain shape. Altered enzymatic activity is crucial to compensate for the lowered enzyme quality under heat stress. © 2024 Society of Chemical Industry.

Engineered Chlorella vulgaris improves bioethanol production and promises prebiotic application.

Microalgal biomass for biofuel production, integration into functional food, and feed supplementation has generated substantial interest worldwide due to its high growth rate, non-competitiveness for agronomic land, ease of cultivation in containments, and presence of several bioactive molecules. In this study, genetic engineering tools were employed to develop transgenic lines of freshwater microalga Chlorella vulgaris with a higher starch content, by up-regulating ADP-glucose pyrophosphorylase (AGPase), which is a rate-limiting enzyme in starch biosynthesis. Expression of the Escherichia coli glgC (AGPase homolog) gene in C. vulgaris led to an increase in total carbohydrate content up to 45.1% (dry cell weight, DCW) in the transgenic line as compared to 34.2% (DCW) in the untransformed control. The starch content improved up to 16% (DCW) in the transgenic alga compared to 10% (DCW) in the control. However, the content of total lipid, carotenoid, and chlorophyll decreased differentially in the transgenic lines. The carbohydrate-rich biomass from the transgenic algal line was used to produce bioethanol via yeast fermentation, which resulted in a higher ethanol yield of 82.82mg/L as compared to 54.41mg/L from the untransformed control. The in vitro digestibility of the transgenic algal starch revealed a resistant starch content of up to 7% of total starch. Faster growth of four probiotic bacterial species along with a lowering of the pH of the growth medium indicated transgenic alga to exert a positive prebiotic effect. Taken together, the study documents the utilization of genetically engineered C. vulgaris with enriched carbohydrates as bioethanol feedstock and functional food ingredients.

As(III)-oxidizing and plant growth-promoting bacteria increase the starch biosynthesis-related enzyme activity, 2-AP levels, and grain quality of arsenic-stressed rice plants

BackgroundGrain quality is an important index of rice production, particularly when plants are grown under stress. Arsenic (As) contamination in paddy fields severely affects rice grain yield and quality. Here, the effects of As and combinations of As(III)-oxidizing bacteria (Pseudomonas stutzeri 4.25, 4.27, and 4.44) and plant growth-promoting bacteria (Delftia acidovorans KKU2500-12 and Cupriavidus taiwanensis KKU2500-3) on enzymes related to starch accumulation in grains and the grain quality of Khao Dawk Mali 105 rice cultivated in As-contaminated soil under greenhouse conditions were investigated.ResultsArsenic affected the activities of starch biosynthesis-related enzymes, and decreases of up to 76.27%, 71.53%, 49.74%, 73.39%, and 47.46% in AGPase, SSS, GBSS, SBE, and SDBE activities, respectively, and 9.42–61.07% in starch accumulation in grains were detected after growth in As-contaminated soil. However, the KKU2500-3/4.25 and KKU2500-3/4.44 combinations yielded the greatest enzyme activities in grains, and compared with the results observed in uninoculated seedlings, increases in starch accumulation of up to 51.16% and 23.81% were found in the inoculated seedlings after growth in medium- and high-As-contaminated soils, at 10–17 and 10–24 days after anthesis, respectively. The bacteria increased the 2-AP content in rice under As stress, possibly via the induction of proline, a 2-AP substrate. Bacterium-inoculated rice had significantly greater 2-AP levels than uninoculated rice, and 2.16–9.93% and 26.57–42.04% increases were detected in rice plants grown in medium- and high-As-contaminated soils, respectively.ConclusionsArsenic toxicity can be mitigated in rice growing under greenhouse conditions by maintaining starch biosynthesis, accumulating amylose, and increasing 2-AP content. The effectiveness of these bacteria should be validated in paddy fields; hence, safe rice grains with a good starch content and aroma could be produced.

Transcriptome and metabolome analyses reveal regulatory networks associated with nutrition synthesis in sorghum seeds

Cereal seeds are vital for food, feed, and agricultural sustainability because they store and provide essential nutrients to human and animal food and feed systems. Unraveling molecular processes in seed development is crucial for enhancing cereal grain yield and quality. We analyze spatiotemporal transcriptome and metabolome profiles during sorghum seed development in the inbred line ‘BTx623’. Morphological and molecular analyses identify the key stages of seed maturation, specifying starch biosynthesis onset at 5 days post-anthesis (dpa) and protein at 10 dpa. Transcriptome profiling from 1 to 25 dpa reveal dynamic gene expression pathways, shifting from cellular growth and embryo development (1–5 dpa) to cell division, fatty acid biosynthesis (5–25 dpa), and seed storage compounds synthesis in the endosperm (5–25 dpa). Network analysis identifies 361 and 207 hub genes linked to starch and protein synthesis in the endosperm, respectively, which will help breeders enhance sorghum grain quality. The availability of this data in the sorghum reference genome line establishes a baseline for future studies as new pangenomes emerge, which will consider copy number and presence-absence variation in functional food traits.

The biosynthesis of storage reserves and auxin is coordinated by a hierarchical regulatory network in maize endosperm.

Grain filling in maize (Zea mays) is intricately linked to cell development, involving the regulation of genes responsible for the biosynthesis of storage reserves (starch, proteins, and lipids) and phytohormones. However, the regulatory network coordinating these biological functions remains unclear. In this study, we identified 1744 high-confidence target genes co-regulated by the transcription factors (TFs) ZmNAC128 and ZmNAC130 (ZmNAC128/130) through chromatin immunoprecipitation sequencing coupled with RNA-seq analysis in the zmnac128/130 loss-of-function mutants. We further constructed a hierarchical regulatory network using DNA affinity purification sequencing analysis of downstream TFs regulated by ZmNAC128/130. In addition to target genes involved in the biosynthesis of starch and zeins, we discovered novel target genes of ZmNAC128/130 involved in the biosynthesis of lipids and indole-3-acetic acid (IAA). Consistently, the number of oil bodies, as well as the contents of triacylglycerol, and IAA were significantly reduced in zmnac128/130. The hierarchical regulatory network centered by ZmNAC128/130 revealed a significant overlap between the direct target genes of ZmNAC128/130 and their downstream TFs, particularly in regulating the biosynthesis of storage reserves and IAA. Our results indicated that the biosynthesis of storage reserves and IAA is coordinated by a multi-TFs hierarchical regulatory network in maize endosperm.

Carbon usage in yellow-fleshed Manihot esculenta storage roots shifts from starch biosynthesis to cell wall and raffinose biosynthesis via the myo-inositol pathway.

Cassava is a crucial staple crop for smallholder farmers in tropical Asia and Sub-Saharan Africa. Although high yield remains the top priority for farmers, the significance of nutritional values has increased in cassava breeding programs. A notable negative correlation between provitamin A and starch accumulation poses a significant challenge for breeding efforts. The negative correlation between starch and carotenoid levels in conventional and genetically modified cassava plants implies the absence of a direct genomic connection between the two traits. The competition among various carbon pathways seems to account for this relationship. In this study, we conducted a thorough analysis of 49 African cassava genotypes with varying levels of starch and provitamin A. Our goal was to identify factors contributing to differential starch accumulation. Considering carotenoid levels as a confounding factor in starch production, we found that yellow- and white-fleshed storage roots did not differ significantly in most measured components of starch or de novo fatty acid biosynthesis. However, genes and metabolites associated with myo-inositol synthesis and cell wall polymer production were substantially enriched in high provitamin A genotypes. These results indicate that yellow-fleshed cultivars, in comparison to their white-fleshed counterparts, direct more carbon toward the synthesis of raffinose and cell wall components. This finding is underlined by a significant rise in cell wall components measured within the 20 most contrasting genotypes for carotenoid levels. Our findings enhance the comprehension of the biosynthesis of starch and carotenoids in the storage roots of cassava.

Insights into starch synthesis and amino acid composition of common buckwheat in response to phosphate fertilizer management strategies

To investigate the response and the regulatory mechanism of common buckwheat starch, amylose, and amylopectin biosynthesis to P management strategies, field experiments were conducted in 2021 and 2022 using three phosphorus (P) levels. Results revealed that the application of 75 kg hm−2 phosphate fertilizer significantly enhanced amylopectin and total starch content in common buckwheat, leading to improved grain weight and starch yield, and decreased starch granule size. The number of upregulated differentially expressed proteins induced by phosphate fertilizer increased with the application rate, with 56 proteins identified as shared differential proteins between different P levels, primarily associated with carbohydrate and amino acid metabolism. Phosphate fertilizer inhibited amylose synthesis by downregulating granule-bound starch synthase protein expression and promoted amylopectin accumulation by upregulating 1,4-alpha-glucan branching enzyme and starch synthase proteins expression. Additionally, Phosphate fertilizer primarily promoted the accumulation of hydrophobic and essential amino acids. These findings elucidate the mechanism of P-induced starch accumulation and offer insights into phosphate fertilizer management and high-quality cultivation of common buckwheat.

Hot Water Treatment Preserves Chinese Chestnut (Castanea mollissima Blume) Quality during Storage by Increasing Its Sugar Accumulation and ROS-Scavenging Ability

Heat treatment is a widely used physical technology for postharvest fruit and crops. The Chinese chestnut cultivar “Kuili” has high sugar and amylose contents, and is popular among people. However, the chestnut quality decreases quickly after harvest. In order to maintain the chestnuts’ quality during storage, this study explores five hot water treatments for chestnuts: T1 (control, no treatment), T2 (50 °C), T3 (65 °C), T4 (75 °C), and T5 (90 °C) for 45 min. T1 was dried at ambient temperature, while the other heat treatments were dried at 30 °C for 30 min. After treatment, chestnuts were placed in plastic trays, covered with a 15 μm thick PVC film, and stored at 4 °C with 70% relative humidity; they remained in the same air for 120 days. Results indicated that T3 and T4 showed slight color changes while maintaining shell and kernel firmness, and their weight loss was reduced (+5–8%), as well as their decay rate (limited to within 20%). The T3, T4, and T5 treatments (from days 60 to 120) decreased their pest survival rates to <2%. Additionally, heat treatments facilitated the accumulation of total soluble sugar and increased the expression of sugar biosynthesis-related genes. Meanwhile, T3 and T4 delayed starch reduction (they maintained relatively higher contents, from 288 to ~320 mg g−1 DW) and altered some starch biosynthesis genes. Furthermore, T2, T3, and T4 exhibited higher antioxidant activity and lower hydrogen peroxide (H2O2) and superoxide anions (O2−) contents than T1. At the end of storage, the scores of T3 and T4 treatments were 55.1 and 52.3, and they ranked first and second among the five treatments, respectively. Therefore, these findings provide valuable insights for controlling postharvest losses in chestnuts.

OsNF-YA8 promotes starch accumulation and influences seed traits by positively regulating starch biosynthesis in rice

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.

Multi-omics analysis reveals the pivotal role of phytohormone homeostasis in regulating maize grain water content

Grain water content (GWC) is a key determinant for mechanical harvesting of maize (Zea mays). In our previous research, we identified a quantitative trait locus, qGWC1, associated with GWC in maize. Here, we examined near-isogenic lines (NILs) NILL and NILH that differed at the qGWC1 locus. Lower GWC in NILL was primarily attributed to reduced grain water weight (GWW) and smaller fresh grain size, rather than the accumulation of dry matter. The difference in GWC between the NILs became more pronounced approximately 35 d after pollination (DAP), arising from a faster dehydration rate in NILL. Through an integrated analysis of the transcriptome, proteome, and metabolome, coupled with an examination of hormones and their derivatives, we detected a marked decrease in JA, along with an increase in cytokinin, storage forms of IAA (IAA-Glu, IAA-ASP), and IAA precursor IPA in immature NILL kernels. During kernel development, genes associated with sucrose synthases, starch biosynthesis, and zein production in NILL, exhibited an initial up-regulation followed by a gradual down-regulation, compared to those in NILH. This discovery highlights the crucial role of phytohormone homeostasis and genes related to kernel development in balancing GWC and dry matter accumulation in maize kernels.

The activation of Chlamydomonas reinhardtii alpha amylase 2 by glutamine requires its N-terminal aspartate kinase-chorismate mutase-tyrA (ACT) domain.

The coordination of assimilation pathways for all the elements that make up cellular components is a vital task for every organism. Integrating the assimilation and use of carbon (C) and nitrogen (N) is of particular importance because of the high cellular abundance of these elements. Starch is one of the most important storage polymers of photosynthetic organisms, and a complex regulatory network ensures that biosynthesis and degradation of starch are coordinated with photosynthetic activity and growth. Here, we analyzed three starch metabolism enzymes of Chlamydomonas reinhardtii that we captured by a cyclic guanosine monophosphate (cGMP) affinity chromatography approach, namely, soluble starch synthase STA3, starch-branching enzyme SBE1, and α-amylase AMA2. While none of the recombinant enzymes was directly affected by the presence of cGMP or other nucleotides, suggesting an indirect binding to cGMP, AMA2 activity was stimulated in the presence of L-glutamine (Gln). This activating effect required the enzyme's N-terminal aspartate kinase-chorismate mutase-tyrA domain. Gln is the first N assimilation product and not only a central compound for the biosynthesis of N-containing molecules but also a recognized signaling molecule for the N status. Our observation suggests that AMA2 might be a means to coordinate N and C metabolism at the enzymatic level, increasing the liberation of C skeletons from starch when high Gln levels signal an abundance of assimilated N.

LAZY4 acts additively with the starch–statolith-dependent gravity-sensing pathway to regulate shoot gravitropism and tiller angle in rice

Rice tiller angle is a key agronomic trait that has significant effects on the establishment of a high-yield rice population. However, the molecular mechanism underlying the control of rice tiller angle remains to be clarified. Here, we characterized the novel tiller-angle gene LAZY4 (LA4) in rice through map-based cloning. LA4 encodes a C3H2C3-type RING zinc-finger E3 ligase localized in the nucleus, and an in vitro ubiquitination assay revealed that the conserved RING finger domain is essential for its E3 ligase activity. We found that expression of LA4 can be induced by gravistimulation and that loss of LA4 function leads to defective shoot gravitropism caused by impaired asymmetric auxin redistribution upon gravistimulation. Genetic analysis demonstrated that LA4 acts in a distinct pathway from the starch biosynthesis regulators LA2 and LA3, which function in the starch–statolith-dependent pathway. Further genetic analysis showed that LA4 regulates shoot gravitropism and tiller angle by acting upstream of LA1 to mediate lateral auxin transport upon gravistimulation. Our studies reveal that LA4 regulates shoot gravitropism and tiller angle upstream of LA1 through a novel pathway independent of the LA2–LA3-mediated gravity-sensing mechanism, providing new insights into the rice tiller-angle regulatory network.

Identification and expression of gene loci underlying the quantitative trait loci of starch content in cassava

ABSTRACT In this study, a genetic linkage map was developed based on simple sequence repeat (SSR) markers to aid in the identification and expression analysis of gene loci underlying the quantitative trait loci (QTL) of starch content (SC) in cassava (Manihot esculenta). A total of 912 polymorphic loci from 4,515 tested SSRs were mapped into 18 linkage groups. QTL analysis revealed four loci with highly significant associations with starch. Gene annotation was performed, and potential genes related to starch synthesis were identified. Of these, expression of four genes related to starch synthesis (Raffinose synthase [RS], Phosphoenolpyruvate kinase [PK], Glucose-6-phosphate [G6P], and Glycogen synthase kinase 3 [GSK-3]) was investigated by real-time PCR assays. The results found a statistically significant difference (p < 0.05) at 3 months after planting (MAP) in all the tested genes, whereas a higher expression was detected in the low-SC group with specific genotypes of linked markers. A higher level of expression was observed in the RS, PK, and G6P genes at six and nine MAP, indicating that starch accumulation mainly occurs after 3 MAP, during which the root system begins to form. The potential genes identified in this study could provide a better understanding of the starch biosynthesis mechanism, and could be useful for applications in marker-assisted selection to facilitate cassava breeding programs in the future.

Genetic elements controlling starch biosynthesis and secondary growth in root and tuber crops

Genetic elements controlling starch biosynthesis and secondary growth in root and tuber crops

Transcriptomics and starch biosynthesis analysis in leaves and developing seeds of mung bean provide a basis for genetic engineering of starch composition and seed quality.

Mung bean starch is distinguished by its exceptional high amylose content and regulation of starch biosynthesis in leaves and storage tissues, such as seeds, share considerable similarities. Genetic engineering of starch composition and content, requires detailed knowledge of starch biosynthetic gene expression and enzymatic regulation. In this study we applied detailed transcriptomic analyses to unravel the global differential gene expression patterns in mung bean leaves and in seeds during various stages of development. The objective was to identify candidate genes and regulatory mechanisms that may enable generation of desirable seed qualities through the use of genetic engineering. Notable differences in gene expression, in particular low expression of the Protein Targeting to Starch (PTST), starch synthase (SS) 3, and starch branching enzyme1 (SBE1) encoding genes in developing seeds as compared to leaves were evident. These differences were related to starch molecular structures and granule morphologies. Specifically, the starch molecular size distribution at different stages of seed development correlated with the starch biosynthesis gene expression of the SBE1, SS1, granule-bound starch synthases (GBSS) and isoamylase 1 (ISA1) encoding genes. Furthermore, putative hormonal and redox controlled regulation were observed, which may be explained by abscisic acid (ABA) and indole-3-acetic acid (IAA) induced signal transduction, and redox regulation of ferredoxins and thioredoxins, respectively. The morphology of starch granules in leaves and developing seeds were clearly distinguishable and could be correlated to differential expression of SS1. Here, we present a first comprehensive transcriptomic dataset of developing mung bean seeds, and combined these findings may enable generation of genetic engineering strategies of for example starch biosynthetic genes for increasing starch levels in seeds and constitute a valuable toolkit for improving mung bean seed quality.