Plant-based proteins: advances in their sources, digestive profiles in vitro and potential health benefits.

Plant-based proteins (PBPs), which are environmentally friendly and sustainable sources of nutrition, can address the emerging challenges facing the global food supply due to the rapidly increasing population. PBPs have received much attention in recent decades as a result of high nutritional values, good functional properties, and potential health effects. This review aims to summarize the nutritional, functional and digestive profiles of PBPs, the health effects of their hydrolysates, as well as processing methods to improve the digestibility of PBPs. The diversity of plant protein sources plays an important role in improving the PBPs quality. Several types of models such as in vitro (the static and semi-dynamic INFOGEST) and in silico models have been proposed and used in simulating the digestion of PBPs. Processing methods including germination, fermentation, thermal and non-thermal treatment can be applied to improve the digestibility of PBPs. PBPs and their hydrolysates show potential health effects including antioxidant, anti-inflammatory, anti-diabetic, anti-hypertensive and anti-cancer activities. Based on the literature, diverse PBPs are ideal protein sources, and exhibit favorable digestive properties and health benefits that could be further improved by different processing technologies. Future research should explore the molecular mechanisms underlying the bioactivity of PBPs and their hydrolysates.

Starch chain-length distributions determine cooked foxtail millet texture and starch physicochemical properties.

Starch chain-length distributions play an important role in controlling cereal product texture and starch physicochemical properties. Cooked foxtail millet texture and starch physicochemical properties were investigated and correlated with starch chain-length distributions in eight foxtail millet varieties. The average chain lengths of amylopectin and amylose were in the range of DP 24-25 and DP 878-1128, respectively. The percentage of short amylopectin chains (Ap1) was negatively correlated with hardness but positively correlated with adhesiveness and cohesion. Conversely, the amount of amylose intermediate chains was positively correlated with hardness but negatively correlated with adhesiveness and cohesion. Additionally, the amount of amylose long chains was negatively correlated with adhesiveness and chewiness. The relative crystallinity (RC) of starch decreased with reductions in the length of amylopectin short chains in foxtail millet. Pasting properties were mainly influenced by the relative length of amylopectin side chains and the percentage of long amylopectin branches (Ap2). Longer amylopectin long chains resulted in lower gelatinization temperature and enthalpy (ΔH). The amount of starch branched chains had important effects on the gelatinization temperature range (ΔT). These results can provide guidance for breeders and food scientists in the selection of foxtail millet with improved quality properties.

Individual or mixing extrusion of Tartary buckwheat and adzuki bean: Effect on quality properties and starch digestibility of instant powder.

Introduction: Tartary buckwheat and adzuki bean, which are classified as coarse grain, has attracted increasing attention as potential functional ingredient or food source because of their high levels of bioactive components and various health benefits. Methods: This work investigated the effect of two different extrusion modes including individual extrusion and mixing extrusion on the phytochemical compositions, physicochemical properties and in vitro starch digestibility of instant powder which consists mainly of Tartary buckwheat and adzuki bean flour. Results: Compared to mixing extrusion, instant powder obtained with individual extrusion retained higher levels of protein, resistant starch, polyphenols, flavonoids and lower gelatinization degree and estimated glycemic index. The α-glucosidase inhibitory activity (35.45%) of the instant powder obtained with individual extrusion was stronger than that obtained with mixing extrusion (26.58%). Lower levels of digestibility (39.65%) and slower digestion rate coefficient (0.25 min-1) were observed in the instant powder obtained with individual extrusion than in mixing extrusion (50.40%, 0.40 min-1) by logarithm-of-slope analysis. Moreover, two extrusion modes had no significant impact on the sensory quality of instant powder. Correlation analysis showed that the flavonoids were significantly correlated with physicochemical properties and starch digestibility of the instant powder. Discussion: These findings suggest that the instant powder obtained with individual extrusion could be used as an ideal functional food resource with anti-diabetic potential.

Transcriptomic analysis of ncRNA and mRNA interactions during leaf senescence in tomato.

An increasing number of non-coding RNAs (ncRNAs) have been discovered recently with the advance of RNA-seq. Nevertheless, the function of ncRNAs in leaf senescence was not fully elucidated. In this study, the whole transcriptome sequencing was employed to characterize the expression profiles of mRNA, lncRNA and miRNA during leaf senescence. A total of 2774 mRNAs, 160 lncRNAs and 117 miRNAs were identified to be significantly differentially expressed between the senescent and the young leaves. Co-expression analysis showed that 160 differential expressing (DE) lncRNAs potentially regulated 946 protein-coding genes in trans, but only 32 targeted protein-coding genes were predicated to be regulated by 30 lncRNAs in cis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of these trans- and cis-target genes revealed that the DE lncRNAs participated in pathways, such as photosynthesis, transporters and circadian rhythm. Furthermore, virus-induced gene silencing (VIGS) was employed to illuminate the role of lncRNAs. The silence of MSTRG.16920 and MSTRG.7613, two intergenic lncRNAs, significantly inhibited leaf senescence induced by darkness, presumably attributed to the downregulated expression of their corresponding target genes Solyc02g069960 and Solyc06g050440, respectively. Notably, Solyc02g069960 and Solyc06g050440 encoding senescence-related NAC transcription factor and reactive oxygen species (ROS)-related peroxidase, respectively, served as positive regulators of leaf senescence. Collectively, our study provides a comprehensive expression profile of lncRNAs in senescent leaf with the concurrent integrated expression of mRNAs and miRNAs.

Mitogen-activated protein kinase 4 is obligatory for late pollen and early fruit development in tomato.

Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules regulating vegetative and reproductive development of plants. However, the molecular mechanisms of the SlMPK4 gene in tomato pollen and fruit development remain elusive. SlMPK4 is preferentially and highly expressed in tomato stamens and its mRNA levels increase during early flower development, peaking at the mature pollen stage. Either up- or downregulation of SlMPK4 expression had no significant effect on tomato vegetative growth. However, RNAi-mediated suppression of SlMPK4 caused defects in pollen development, resulting in pollen abortion. The aborted pollen grains were either malformed or collapsed and completely lacked viability, resulting in a predominantly reduced fruit set rate in RNAi lines compared with control and overexpressing transgenic plants. Interestingly, seed development was inhibited in RNAi lines. Moreover, >12% of emasculated RNAi flowers developed seedless fruits without pollination. Anthers can produce typical microspore mother cells as well as uninucleate microspores, according to cytological investigations, while binucleate pollen ceased to produce typical mature pollen. Pollen abortion was further confirmed by transmission electron microscopy analysis at the binucleate stage in RNAi plants. The exine layer in aberrant pollen had a normal structure, while the intine layer appeared thicker. Suppression of SlMPK4 affects the transcript level of genes related to cell wall formation and modification, cell signal transduction, and metabolic and biosynthetic processes. A subset of genes that may be putative substrates of plant MAPKs were also differentially changed in RNAi transgenic flowers. Taken together, these results suggest that SlMPK4 plays a critical role in regulating pollen development and fruit development in tomato plants.

Phytochrome interacting factor 3 regulates pollen mitotic division through auxin signalling and sugar metabolism pathways in tomato.

The development of viable pollen determines male fertility, and is crucial for reproduction in flowering plants. Phytochrome interacting factor 3 (PIF3) acts as a central regulator of plant growth and development, but its relationship with pollen development has not been determined. Through genetic, histological and transcriptomic analyses, we identified an essential role for SlPIF3 in regulating tomato (Solanum lycopersicum) pollen development. Knocking out SlPIF3 using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 resulted in pollen mitosis I arrest, and a failure to form viable pollen. We further demonstrated that both glutamate synthase 1 (SlGLT1) and cell wall invertase 9 (SlCWIN9), involved in auxin and sugar homeostasis, respectively, colocalised with SlPIF3 in the anthers and were directly regulated by SlPIF3. Knockout of either SlGLT1 or SlCWIN9 phenocopied the pollen phenotype of SlPIF3 knockout (Slpif3) lines. Slpif3 fertility was partially restored by exogenous auxin indole-3-acetic acid in a dose-dependent manner. This study reveals a mechanism by which SlPIF3 regulates pollen development and highlights a new strategy for creating hormone-regulated genic male sterile lines for tomato hybrid seed production.

RNA N6-Methyladenosine Responds to Low-Temperature Stress in Tomato Anthers.

Cold stress is a serious threat to subtropical crop pollen development and induces yield decline. N6-methyladenosine (m6A) is the most frequent mRNA modification and plays multiple physiological functions in plant development. However, whether m6A regulates pollen development is unclear, and its putative role in cold stress response remains unknown. Here, we observed that moderate low-temperature (MLT) stress induced pollen abortion in tomato. This phenotype was caused by disruption of tapetum development and pollen exine formation, accompanied by reduced m6A levels in tomato anther. Analysis of m6A-seq data revealed 1,805 transcripts displayed reduced m6A levels and 978 transcripts showed elevated m6A levels in MLT-stressed anthers compared with those in anthers under normal temperature. These differentially m6A enriched transcripts under MLT stress were mainly related to lipid metabolism, adenosine triphosphatase (ATPase) activity, and ATP-binding pathways. An ATP-binding transcript, SlABCG31, had significantly upregulated m6A modification levels, which was inversely correlated to the dramatically downregulated expression level. These changes correlated with higher abscisic acid (ABA) levels in anthers and disrupted pollen wall formation under low-temperature stress. Our findings characterized m6A as a novel layer of complexity in gene expression regulation and established a molecular link between m6A methylation and tomato anther development under low-temperature conditions.

PIF4 negatively modulates cold tolerance in tomato anthers via temperature-dependent regulation of tapetal cell death.

Extreme temperature conditions seriously impair male reproductive development in plants; however, the molecular mechanisms underlying the response of anthers to extreme temperatures remain poorly described. The transcription factor phytochrome-interacting factor4 (PIF4) acts as a hub that integrates multiple signaling pathways to regulate thermosensory growth and architectural adaptation in plants. Here, we report that SlPIF4 in tomato (Solanum lycopersicum) plays a pivotal role in regulating cold tolerance in anthers. CRISPR (clustered regularly interspaced short palindromic repeats)-associated nuclease Cas9-generated SlPIF4 knockout mutants showed enhanced cold tolerance in pollen due to reduced temperature sensitivity of the tapetum, while overexpressing SlPIF4 conferred pollen abortion by delaying tapetal programmed cell death (PCD). SlPIF4 directly interacts with SlDYT1, a direct upstream regulator of SlTDF1, both of which (SlDYT1 and SlTDF1) play important roles in regulating tapetum development and tapetal PCD. Moderately low temperature (MLT) promotes the transcriptional activation of SlTDF1 by the SlPIF4-SlDYT1 complex, resulting in pollen abortion, while knocking out SlPIF4 blocked the MLT-induced activation of SlTDF1. Furthermore, SlPIF4 directly binds to the canonical E-box sequence in the SlDYT1 promoter. Collectively, these findings suggest that SlPIF4 negatively regulates cold tolerance in anthers by directly interacting with the tapetal regulatory module in a temperature-dependent manner. Our results shed light on the molecular mechanisms underlying the adaptation of anthers to low temperatures.

The Arabidopsis SMALL AUXIN UP RNA32 Protein Regulates ABA-Mediated Responses to Drought Stress.

SMALL AUXIN UP-REGULATED RNAs (SAURs) are recognized as auxin-responsive genes involved in the regulation of abiotic stress adaptive growth. Among the growth-limiting factors, water-deficit condition significantly affects plant growth and development. The putative function of SAUR family member AtSAUR32 has the potential to diminish the negative impact of drought stress, but the exact function and mode of action remain unclear in Arabidopsis. In the current study, AtSAUR32 gene was cloned and functionally analyzed. AtSAUR32 localized to the plasma membrane and nucleus was dominantly expressed in roots and highly induced by abscisic acid and drought treatment at certain time points. The stomatal closure and seed germination of saur32 were less sensitive to ABA relative to AtSAUR32-overexpressed line (OE32-5) and wild type (WT). Moreover, the saur32 mutant under drought stress showed increased ion leakage while quantum yield of photosystem II (ΦPSII) and endogenous ABA accumulation were reduced, along with the expression pattern of ABA/stress-responsive genes compared with WT and the OE32-5 transgenic line. Additionally, yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays showed that AtSAUR32 interacted with clade-A PP2C proteins (AtHAI1 and AtAIP1) to regulate ABA sensitivity in Arabidopsis. Taken together, these results indicate that AtSAUR32 plays an important role in drought stress adaptation via mediating ABA signal transduction.

Effects of pre-oxidation on residual dissolved aluminum in coagulated water: A pilot-scale study.

Residual dissolved aluminum (Al) in drinking water is becoming a serious concern due to its high potential risks to human health. However, the mechanism by which residual dissolved Al forms is yet to be elucidated in detail. In this study, the effects of pre-oxidation by ozonation and chlorination on the properties of dissolved organic matter (DOM) and residual Al concentrations remaining in solution after coagulation were explored in a pilot-scale test. Changes in the DOM properties caused by the water treatment process were characterized by ultraviolet-visible absorbance spectroscopy. Theprotonation-active sites, carboxylic- and phenolic-type groupsof DOM were quantified by spectral parameter DlnA400 (differential log-transformed spectra at wavelength 400 nm) in combination with the revised non-ideal competitive adsorption model. The results show that ozonation and chlorination significantly affect the properties of DOM and the amount of residual dissolved Al in coagulated drinking water. This effect was associated with the changes in the carboxylic- and phenolic-type groups in DOM. Results of the study show that residual dissolved Al in coagulated water can be controlled by affecting theAl binding sites in DOM by pre-oxidation before coagulation. The nature of pre-oxidation agent and its dosage should be selected depending on the quality of the raw water to be treated. Ozonation was concluded to be preferable pre-oxidation agent for the water in examined this study.