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.

Ultrasound-assisted annealing treatment to improve physicochemical and digestive properties of banana flour.

BACKGROUND: Banana flour can provide a solution to people with gluten intolerance, as it is gluten-free. Native banana flour may have limited functionality in certain applications. In this study, banana flour was modified by ultrasonic (US) and annealing (ANN) treatments at four incubation time spans, namely 12, 24, 36 and 72 h, separately or combined sequentially (US-ANN) to enhance the physicochemical and digestive properties. RESULTS: US led to exposed granular surfaces and damaged non-starch components. Both treatments, at extended incubation time, increased crystallinity, resulting in a narrower starch gelatinization temperature range. The swelling power was significantly lower for ANN and US-ANN compared to US alone, providing a delay of gelatinization temperature. However, none of the treatments affected the gelatinization enthalpy. Furthermore, US increased peak viscosity, breakdown, final viscosity and setback whereas the opposite results were obtained for ANN and US-ANN. Additionally, US prior to ANN significantly increased the resistant starch (RS) content for annealing times over 24 h, especially for the US-ANN treatment for 72 h, which provided the highest RS content (49.3%) compared to ANN treatment for 72 h (44.0%) and native flour (36.3%). CONCLUSIONS: US prior to ANN treatment offers an alternative method to improve the functional and digestive properties of banana flour, extending the range of applications. © 2024 Society of Chemical Industry.

Starch phosphorylation regulates starch granule morphological homogeneity in Arabidopsis thaliana.

Starch granule morphological homogeneity presents a gap in starch research. Transitory starch granules in wild-type plants are discoid, regardless of species. Notably, while the shape of starch granules can differ among mutants, it typically remains homogeneous within a genotype. We found an Arabidopsis thaliana mutant, dpe2sex4, lacking both the cytosolic disproportionating enzyme 2 (DPE2) and glucan phosphatase SEX4, showing an unprecedented bimodal starch granule diameter distribution when grown under a light/dark rhythm. dpe2sex4 contained 2 types of starch granules: large granules and small granules. In contrast to the double starch initiation in wheat (Triticum aestivum) endosperm, where A-type granules are initiated first and B-type granules are initiated later, dpe2sex4 small and large granules developed simultaneously in the same chloroplast. Compared with the large granules, the small granules had more branched amylopectin and less surface starch-phosphate, thus having a more compact structure that may hinder starch synthesis. During plant aging, the small granules barely grew. In in vitro experiments, fewer glucosyl residues were incorporated in small granules. Under continuous light, dpe2sex4 starch granules were morphologically homogeneous. Omitting the dark phase after a 2-wk light/dark cycle by moving plants into continuous light also reduced morphological variance between these 2 types of granules. These data shed light on the impact of starch phosphorylation on starch granule morphology homogeneity.

High-pressure pasting performance and multilevel structures of short-term microwave-treated high-amylose maize starch.

Microwave treatment is an environmentally friendly method for modification of high-amylose maize starch (HAMS). Here, the effects of short-time (≤120 s) microwave treatment on the structure and pasting of two types of HAMSs, Gelose 50 (HAMSI) and Gelose 80 (HAMSII), with apparent amylose content (AAC) of 45 % and 58 %, respectively, was studied using a multiscale approach including X-ray scattering, surface structures, particle size distribution, molecular size distributions and high temperature/pressure Rapid Visco Analysis (RVA)-4800 pasting. As compared to starch with no amylose (waxy maize starch, WMS) and 25 % amylose content (normal maize starch, NMS), HAMSI underwent similar structural and pasting changes as WMS and NMS upon microwave treatment, and it might primarily be attributed to the amylopectin fraction that was affected by cleavage of the connector chains between double helices and backbone chains, which decreased the crystallinity and thickness of the crystalline lamellae. However, the multi-scale structure of HAMSII was almost unaffected by this treatment. The pasting properties of fully gelatinized HAMSI starch showed a decrease in RVA-4800 peak and final viscosities after microwave treatment. In contrast, for HAMSII starch, the microwave treatment led to an increase in these viscosities. The combined results highlight the influence of varying AAC on the effects of microwave-mediated modification, leading to diverse alterations in the structure and functionality of starches.

Strategies for starch customization: Agricultural modification.

Raw starch is commonly modified to enhance its functionality for industrial applications. There is increasing demand for 'green' modified starches from both end-consumers and producers. It is well known that environmental conditions are key factors that determine plant growth and yield. An increasing number of studies suggest growth conditions can expand affect starch structure and functionality. In this review, we summarized how water, heat, high nitrogen, salinity, shading, CO2 stress affect starch biosynthesis and physicochemical properties. We define these treatments as a fifth type of starch modification method - agricultural modification - in addition to chemical, physical, enzymatic and genetic methods. In general, water stress decreases peak viscosity and gelatinization enthalpy of starch, and high temperature stress increases starch gelatinization enthalpy and temperature. High nitrogen increases total starch content and regulates starch viscosity. Salinity stress mainly regulates starch and amylose content, both of which are genotype-dependent. Shading stress and CO2 stress can both increase starch granule size, but these have different effects on amylose content and amylopectin structure. Compared with other modification methods, agricultural modification has the advantage of operating at a large scale and a low cost and can help meet the ever-rising market of clean-label foods and ingredients.

Colorimetric pH indicators based on well-defined amylose and amylopectin matrices enriched with anthocyanins from red cabbage.

This study aimed to prepare a novel colorimetric indicator film from virtually pure (99 %) amylose (AM) and anthocyanins extracted from red cabbage (RCA). The AM used was a unique engineered bulk material extracted from transgenic barley grains. Films produced by solution casting were compared to normal barely starch (NB) and pure barley amylopectin (AP), with amylose contents of 30 % and 0 %, respectively. The pH-indicator films were produced by incorporation of RCA into the different starch support matrices with different amylose contents. Barrier, thermal, and mechanical properties, photo degradation stability, and release behavior data revealed that RCA interact differently through the glucan matrices. Microstructural observations showed that RCA were evenly dispersed in the glucan matrix, and AM+RCA indicator films showed high UV-barrier and mechanical performance over normal starch. FTIR revealed that RCA was properly affected by the AM matrix. Moreover, the AM+RCA films showed sensitive color changes in the pH range (2-11) and a predominant Fickian diffusion release mechanism for RCA. This study provides for the first time data regarding AM films with RCA and their promising potential for application as support matrices in responsive food and other industrial biodegradable packaging materials.

Climate-Nutrient-Crop Model: Novel Insights into Grain-Based Food Quality.

Mineral nutrients spatiotemporally participate in the biosynthesis and accumulation of storage biopolymers, which directly determines the harvested grain yield and quality. Optimizing fertilizer nutrient availability improves the grain yield, but quality aspects are often underestimated. We hypothesize that extensive mineral nutrients have significant effects on the biosynthesis, content, and composition of storage proteins, ultimately determining physicochemical properties and food quality, particularly in the context of climate change. To investigate this, we hierarchized 16 plant mineral nutrients and developed a novel climate-nutrient-crop model to address the fundamental question of the roles of protein and starch in grain-based food quality. Finally, we recommend increasing the added value of mineral nutrients as a socioeconomic strategy to enhance agro-food profitability, promote environmental sustainability, and improve climate resilience.

Periodic changes in chain lengths distribution parameters of wheat starch during endosperm development.

This study analyzed the molecular structure of developing wheat endosperm starch at different stages after anthesis (DAA) using chain length distribution analysis by size exclusion chromatography (SEC) and fluorophore-assisted carbohydrate electrophoresis. Our results revealed periodic changes in the content of both amylose and amylopectin fractions. Specifically, the content of amylose chains with a degree of polymerization (DP) > 100 significantly decreased from 5 to 10 DAA (28% to 21%) and from 15 to 20 DAA (29% to 26%), but increased between 10 and 15 DAA (21% to 29%) and 20 to 25 DAA (30.0% to 33%). Conversely, the content of short amylopectin chains with DP ≤ 32 showed the opposite trend. Interestingly, mRNA expression levels of key starch biosynthesis genes did not exhibit periodic changes. These findings contribute to our understanding of starch biosynthesis and provide important insights for the development of starch-based products.

Improved Hydrolysis of Granular Starches by a Psychrophilic α-Amylase Starch Binding Domain-Fusion.

Degradation of starch granules by a psychrophilic α-amylase, AHA, from the Antarctic bacterium Pseudoalteromonas haloplanktis TAB23 was facilitated by C-terminal fusion to a starch-binding domain (SBD) from either Aspergillus niger glucoamylase (SBDGA) or Arabidopsis thaliana glucan, water dikinase 3 (SBDGWD3) via a decapeptide linker. Depending on the waxy, normal or high-amylose starch type and the botanical source, the AHA-SBD fusion enzymes showed up to 3 times higher activity than AHA wild-type. The SBD-fusion thus increased the density of enzyme attack-sites and binding-sites on the starch granules by up to 5- and 7-fold, respectively, as measured using an interfacial catalysis approach that combined conventional Michaelis-Menten kinetics, with the substrate in excess, and inverse kinetics, having enzyme in excess, with enzyme-starch granule adsorption isotherms. Higher substrate affinity of the SBDGA compared to SBDGWD3 was accompanied by the superior activity of AHA-SBDGA in agreement with the Sabatier principle of adsorption limited heterogenous catalysis.

Interfacial Catalysis during Amylolytic Degradation of Starch Granules: Current Understanding and Kinetic Approaches.

Enzymatic hydrolysis of starch granules forms the fundamental basis of how nature degrades starch in plant cells, how starch is utilized as an energy resource in foods, and develops efficient, low-cost saccharification of starch, such as bioethanol and sweeteners. However, most investigations on starch hydrolysis have focused on its rates of degradation, either in its gelatinized or soluble state. These systems are inherently more well-defined, and kinetic parameters can be readily derived for different hydrolytic enzymes and starch molecular structures. Conversely, hydrolysis is notably slower for solid substrates, such as starch granules, and the kinetics are more complex. The main problems include that the surface of the substrate is multifaceted, its chemical and physical properties are ill-defined, and it also continuously changes as the hydrolysis proceeds. Hence, methods need to be developed for analyzing such heterogeneous catalytic systems. Most data on starch granule degradation are obtained on a long-term enzyme-action basis from which initial rates cannot be derived. In this review, we discuss these various aspects and future possibilities for developing experimental procedures to describe and understand interfacial enzyme hydrolysis of native starch granules more accurately.

Recent advances on bioactive compounds, biosynthesis mechanism, and physiological functions of Nelumbo nucifera.

Nelumbo nucifera Gaertn, commonly known as lotus, is a genus comprising perennial and rhizomatous aquatic plants, found throughout Asia and Australia. This review aimed to cover the biosynthesis of flavonoids, alkaloids, and lipids in plants and their types in different parts of lotus. This review also examined the physiological functions of bioactive compounds in lotus and the extracts from different organs of the lotus plant. The structures and identities of flavonoids, alkaloids, and lipids in different parts of lotus as well as their biosynthesis were illustrated and updated. In the traditional medicine systems and previous scientific studies, bioactive compounds and the extracts of lotus have been applied for treating inflammation, cancer, liver disease, Alzheimer's disease, etc. We suggest future studies to be focused on standardization of the extract of lotus, and their pharmacological mechanisms as drugs or functional foods. This review is important for the lotus-based food processing and application.

Starch granular size and multi-scale structure determine population patterns in bivariate flow cytometry sorting.

Bivariate flow cytometry (FC) sorting with forward scatter (FSC) and side scatter (SSC) is a recently established novel technique to separate starch granules. However, the forming mechanism of starch FC-dependent population patterns (i.e. the number of subgroups (NS) and FSC/SSC-dependent distribution patterns) remain partly elusive. For this, the correlation of granular size and multi-scale structure of native starches and FC-dependent population patterns was investigated through employing a wide range of native starches originating from different species involving cereal-, pulse-, and tuber crops. Results showed NS was pertinent with particle size, amylose content (AC), amylopectin chains length distribution, lamellar structure, short-range ordered structure. The distinct NS was determined by impacts of native starch FSC / SSC-dependent distribution patterns. Specifically, starch granular size significantly correlated with both FSC and SSC-dependent distribution patterns. The proportion of chains with DP 6-12 was the intra-molecular decisive factor to influence short-range ordered structure, finally leading to FSC-dependent distribution patterns. By contrast, AC was another intra-molecular index to determine SSC-dependent distribution patterns through affecting lamellar structure and short-range ordered structure.

High-amylose starch: Structure, functionality and applications.

Starch with a high amylose (AM) content (high AM starch, HAS) has attracted increasing research attention due to its industrial application potential, such as functional foods and biodegradable packaging. In the past two decades, HAS structure, functionality, and applications have been the research hotspots. However, a review that comprehensively summarizes these areas is lacking, making it difficult for interested readers to keep track of past and recent advances. In this review, we highlight studies that benefited from rapidly developing techniques, and systematically review the structure, functionality, and applications of HAS. We particularly emphasize the relationships between HAS molecular structure and physicochemical properties.

The effects of different types of high-amylose maize starches on viscosity and digestion of acidified milk gels.

High-amylose maize starch (HAMS) can provide dietary fiber to foods. In this study, we investigated the effects of three HAMSs (Gelose 50, Hylon VII, and NAFU50) on the functionality of casein (CA) and/or whey protein (WP) networks in acidified milk gels using normal maize starch (NMS) as a control thickener. When compared with NMS, HAMSs performed better in increasing the resistant starch content (RS), storage modulus, loss modulus, and complex viscosity of all the milk gels. The results are attributed to the retention of the starch granular integrity of HAMSs during the preparation of the milk gels, as observed by microscopy. HylonVII exhibited the highest RS and viscosity in all milk gel systems, especially in WP gels (71.8 % RS and >3000 Pa.s at 1 Hz viscosity). Our data provide support for the potential of using HAMS to develop healthier yogurt products using functional thickeners from natural sources.

Effects of growth temperature on multi-scale structure of root tuber starch in sweet potato.

Sweet potato was planted at three soil and air temperatures (21, 25 and 28 °C) with the same humidity and light time/intensity. Root tuber starches were isolated, and their multi-scale structures were investigated to reveal the effects of growth temperature on starch properties. Growth temperature did not change the morphology and amylose content of starch, but markedly increased the size of starch from volume-weighted mean diameter 12.2 µm to 17.0 µm. Starch grown at high growth temperature exhibited less A branch-chains and lower branching degree of amylopectin and more B2 and B3+ branch-chains of amylopectin than at low growth temperature. With increasing growth temperature, starch changed from CC-type to CA-type, its relative crystallinity and lamellar peak intensity increased, and the thickness of crystalline and amorphous lamellae did not significantly change. Starch grown at high growth temperature exhibited significantly higher gelatinization temperature than at low growth temperature, but had similar gelatinization enthalpy.

The effects of drought treatments on biosynthesis and structure of maize starches with different amylose content.

We investigated the effects of drought stress (DS) on maize varieties with different amylose content (AC). In starches with AC of 33 %, DS increased the contents of amylopectin (AP) chains with a degree of polymerization (DP) > 36 and decreased the AP chains with DP ≤ 36, while the AC was unchanged. DS decreased the crystallinity, the thickness of both amorphous and crystalline lamellae, and average granular size. In contrast, the digestibility increased. For starches with AC of 45 %, DS increased the content of AP chains with DP > 24 and AC, while the contents of AP chains with DP ≤ 24 decreased. DS produced starch with thinner crystalline lamellae, thicker amorphous lamellae, more elongated and larger granules. The digestibility of the starches decreased. In starches with AC of 53 %, moderate DS led to similar structural and functional changes as found for starches with AC of 45 %. Finally, severe DS resulted in the decrease of AC.

Corrigendum: Strategies for Efficient Gene Editing in Protoplasts of Solanum tuberosum Theme: Determining gRNA Efficiency Design by Utilizing Protoplast (Research).

[This corrects the article DOI: 10.3389/fgeed.2021.795644.].

Polysaccharide-based nanosystems: a review.

Polysaccharide-based nanosystem is an umbrella term for many areas within research and technology dealing with polysaccharides that have at least one of their dimensions in the realm of a few hundreds of nanometers. Nanoparticles, nanocrystals, nanofibers, nanofilms, and nanonetworks can be fabricated from many different polysaccharide resources. Abundance in nature, cellulose, starch, chitosan, and pectin of different molecular structures are widely used to fabricate nanosystems for versatile industrial applications. This review presents the dissolution and modification of polysaccharides, which are influenced by their different molecular structures and applications. The dissolution ways include conventional organic solvents, ionic liquids, inorganic strong alkali and acids, enzymes, and hydrothermal treatment. Rheological properties of polysaccharide-based nano slurries are tailored for the purpose functions of the final products, e.g., imparting electrostatic functions of nanofibers to reduce viscosity by using lithium chloride and octenyl succinic acid to increase the hydrophobicity. Nowadays, synergistic effects of polysaccharide blends are increasingly highlighted. In particular, the reinforcing effect of nanoparticles, nanocrystals, nanowhiskers, and nanofibers to hydrogels, aerogels, and scaffolds, and the double network hydrogels of a rigid skeleton and a ductile substance have been developed for many emerging issues.

High pressure/temperature pasting and gelling of starch related to multilevel structure-analyzed with RVA 4800.

This paper evaluated the relationship between multi-scale structure and high-pressure gelation properties of nine types of starches with different amylose (AM) content and crystalline polymorphic structure by RVA 4800. Higher average chain lengths of long amylopectin (AP) chains (DP > 36) and AM content, and lower relative content of short AP chains (DP ≤ 36) contributed to the higher peak temperature and peak time at 95-140 °C, and lower peak viscosity, through viscosity, and final viscosity at 95-110 °C. Rheological and texture parameters including storage modulus, loss modulus, hardness, and gumminess, had no significant correlation with starch structural parameters at 95-120 °C, but were instead controlled by AM molecular structure at 130-140 °C. AM content was mainly responsible for the rheological behaviors of starch gels at 130-140 °C, and short and intermediate AM chains were mainly associated with the texture of starch gels at the temperature ranges.