Knowledge Maps and Emerging Trends in Cell-Cultured Meat since the 21st Century Research: Based on Different National Perspectives of Spatial-Temporal Analysis.

Cell-cultured meat holds significant environmental value as an alternative protein source. Throughout the 21st century, cell-cultured meat has progressively penetrated commercial markets. However, a systematic review encompassing the entire field needs improvement. Employing Citespace, Vosviewer, and R-Bibliometrix software, a bibliometric analysis was used to present the research progress and general development trends of 484 articles on cell-cultured meat from 2000 to 2022 based on countries, authors, institutions, and keywords. This analysis provides ideas for the future development of cell-cultured meat in different countries or regions worldwide. Research on cell-cultured meat from 2000 to 2022 has undergone two phases: fluctuating growth (2000-2013) and rapid growth (2013-2022). Noteworthy contributions to cell-cultured meat studies emerge from author groups in the United States of America, the United Kingdom, and China, with influential institutions like the University of Bath significantly impacting pertinent research. Furthermore, over the past two decades, research has leaned towards exploring topics such as "biomaterials", "cultured", "land use", "public opinion", "animal welfare", and "food safety". Furthermore, this study reveals differences in nomenclature between regions and institutions. "Cultured meat" is more popular in some countries than in other forms. Institutions in Asia use "cultured meat" more frequently; however, institutions in the Americas adopt "cultivated meat" and rarely adopt "in vitro meat", and institutions in the European region have no particularly prominent tendency towards a specific nomenclature. Future research should emphasize aligning the labeling of cell-cultured meat with effective management strategies and referencing regulatory policies across various countries. For the first time, we use three different bibliometric methods to analyze temporal and spatial variation in research on cellular meat. The results of this study have a multiplier effect. We provide a theoretical basis and a practical reference for the identification of alternatives in the dual context of "food crisis and food security" and "climate crisis". At the same time, we also provide a reference for the sustainable development of the food system.

How Sugar Labeling Affects Consumer Sugar Reduction: A Case of Sucrose Grade Labels in China.

The effectiveness of sugar labeling depends not only on direct sugar reduction but also on the extent to which compensatory eating occurs. This study focuses on the use of sucrose grade labels in the Chinese market to investigate not only consumers' willingness to pay (WTP) for different sucrose labels but also the consistency of their sugar control behavior when confronted with unlabeled processed foods. The findings reveal that consumers are willing to pay approximately 4%, 7%, and 7% more for yogurt labeled as "low sucrose", "no sucrose", and "no sucrose with sugar substitutes", respectively, compared to yogurt labeled as "regular sucrose." Furthermore, when subsequently presented with unlabeled toast, a significant proportion of consumers who initially chose "no sucrose" yogurt continued to select wholewheat toast, which contains less sugar than white and coconut toast. This indicates their commitment to maintaining their sugar control behavior. The study provides valuable experimental evidence for researchers, food manufacturers, and policymakers regarding the efficacy of sucrose grade labels. In particular, it offers policymakers insights into guiding consumers to promote sustainable healthy diets.

Tomato SlMAPK3 Modulates Cold Resistance by Regulating the Synthesis of Raffinose and the Expression of SlWRKY46.

Mitogen-activated protein kinase (MAPK) cascades and raffinose have been observed to increase in plants exposed to cold. However, it remains elusive whether and how MAPK regulates raffinose synthesis under cold stress. Here, overexpression of SlMAPK3 promoted the accumulation of galactinol and raffinose under cold stress, while CRISPR/Cas9-mediated mutants showed the opposite results. Moreover, SlMAPK3 promoted the expression of SlWRKY46 at low temperatures and interacted with SlWRKY46 protein. Overexpression of SlWRKY46 enhanced cold resistance. Furthermore, SlWRKY46 directly bound to the promoter of SlGols1 to enhance its expression and promoted the accumulation of raffinose. Virus-induced gene-silencing (VIGS)-mediated knockdown of SlGols1 remarkably elevated cold sensitivity and reduced raffinose content. Meanwhile, exogenous supplementation of raffinose could improve the cold tolerance of tomato plants. Thus, our data indicates that SlMAPK3 modulates cold resistance by regulating raffinose content and SlWRKY46 expression. SlWRKY46 also promotes the accumulation of raffinose by inducing the expression of SlGols1.

Recent Advances in Dissecting the Function of Ethylene in Interaction between Host and Pathogen.

Pathogens influence the growth and development of plants, resulting in detrimental damage to their yields and quality. Ethylene, a gaseous phytohormone, serves a pivotal function in modulating diverse physiological processes in plants, including defense mechanisms against pathogen invasion. Ethylene biosynthesis is involved in both plants and pathogens. Recent empirical research elucidates the intricate interactions and regulatory mechanisms between ethylene and pathogens across various plant species. In this review, we provide a comprehensive overview of the latest findings concerning ethylene's role and its regulatory networks in host-pathogen interactions. Additionally, we explore the crosstalk between ethylene and other phytohormones. Points regarding ethylene emission and its modulation by pathogens are also emphasized. Moreover, we also discuss potential unresolved issues in the field that warrant further investigation.

CRISPR/Cas9-Mediated SlATG5 Mutagenesis Reduces the Resistance of Tomato Fruit to Botrytis cinerea.

Tomato fruit is highly susceptible to infection by Botrytis cinerea (B. cinerea), a dominant pathogen, during storage. Recent studies have shown that autophagy is essential for plant defense against biotic and abiotic stresses. Autophagy-related gene 5 (ATG5) plays a key role in autophagosome completion and maturation, and is rapidly induced by B. cinerea, but the potential mechanisms of ATG5 in Solanum lycopersicum (SlATG5) in postharvest tomato fruit resistance to B. cinerea remain unclear. To elucidate the role of SlATG5 in tomato fruit resistant to B. cinerea, CRISPR/Cas9-mediated knockout of SlATG5 was used in this study. The results showed that slatg5 mutants were more vulnerable to B. cinerea and exhibited more severe disease symptoms and lower activities of disease-resistant enzymes, such as chitinase (CHI), ß-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO), than the wild type (WT). Furthermore, the study observed that after inoculation with B. cinerea, the relative expression levels of genes related to salicylic acid (SA) signaling, such as SlPR1, SlEDS1, SlPAD4, and SlNPR1, were higher in slatg5 mutants than in WT. Conversely, the relative expression levels of jasmonic acid (JA) signaling-related genes SlLoxD and SlMYC2 were lower in slatg5 mutants than in WT. These findings suggested that SlATG5 positively regulated the resistance response of tomato fruit to B. cinerea by inhibiting the SA signaling pathway and activating the JA signaling pathway.

Postharvest fruit quality of tomatoes influenced by an ethylene signaling component during long-term cold storage.

Ethylene production is essential for improving cold resistance of postharvest tomatoes. However, the role of ethylene signaling pathway in maintaining fruit quality during long-term cold storage remains poorly understood. Here, we demonstrated that a partial loss of function in ethylene signaling by mutation of Ethylene Response Factor 2 (SlERF2), worsened fruit quality during cold storage, as determined by visual characterization, and physiological analyses of membrane damage and reactive oxygen species metabolism. In addition, the transcriptions of genes related to abscisic acid (ABA) biosynthesis and signaling were also altered by SlERF2 gene in response to cold storage. Furthermore, mutation of SlERF2 gene compromised cold-induced expression of genes in the C-repeat/dehydration-responsive binding factor (CBF) signaling pathway. Therefore, it's concluded that an ethylene signaling component, SlERF2 contributed to the regulations of ABA biosynthesis and signaling, as well as CBF cold signaling pathway, ultimately affecting the fruit quality during long-term cold storage of tomatoes.

SlMAPK3 Positively Regulates the Ethylene Production of Postharvest Tomato Fruits and Is Involved in Ethylene-Mediated Cold Tolerance.

Mitogen-activated protein kinase (MAPK) cascades and ethylene are crucial for plant growth, development, and stress responses, but their potential mechanisms in cold resistance remain unclear. We revealed that SlMAPK3 transcript levels were dramatically induced by cold treatment in an ethylene-dependent manner. Under cold stress, the proline content of SlMAPK3-overexpression fruit was 96.5 and 115.9% higher than that of wild-type fruit (WT), respectively, while the ion leakage was 37.3 and 32.5% lower than that of WT. RNA sequencing revealed that overexpression of SlMAPK3 caused upregulation of genes that are enriched in the ethylene-activated signaling pathway (GO:0009873), cold signaling pathway (GO:0009409), and heat signaling pathway (GO:0009408). RT-qPCR demonstrated that the expression levels of SlACS2, SlACS4, SlSAHH, SlCBF1, SlDREB, SlGolS1, and SlHSP17.7 in the OE.MAPK3 fruits were consistent with the RNA sequencing results. Meanwhile, the knockout of SlMAPK3 reduced the ethylene content, ACC content, and ACS activity. Moreover, the knockout of SlMAPK3 reduced the positive effect of ethylene in cold stress, while suppressing the expression of SlICE1 and SlCBF1. In conclusion, our study demonstrated a novel mechanism by which SlMAPK3 positively regulates the ethylene production of postharvest tomato fruits and is involved in ethylene-mediated cold tolerance.

The role of environmental-related message on consumer acceptance of novel food production technology: an experimental investigation on artificial meat products.

This study investigates the effects of environmental-related message on consumers' acceptance of artificial meat products in China. By focusing on the introduction of plant-based and cultured meat, a novel dataset involving 3441 research participants was collected in China through the online platform with randomized controlled experiments. The results show that for promotion-focused participants, gain-oriented message is effective while avoidance-oriented message is not in improving their acceptance of artificial meat. For prevention-focused participants, both gain- and avoidance-oriented messages are found to increase their acceptance of artificial meat effectively. To quantify the average impacts of environmental-related message and consumers' regulatory focus fit on the acceptance of plant-based meat and cultured meat, a seemingly unrelated regression model is applied. The results reveal that for a randomly selected individual, exposing with both gain- and avoidance-oriented messages would increase his or her acceptance of artificial meat comparing to exposing with neutral message. In addition, we find participants who use promotion focus more tend to have higher acceptance scores for both kinds of artificial meat on average.

Identification of bHLH family genes in Agaricus bisporus and transcriptional regulation of arginine catabolism-related genes by AbbHLH1 after harvest.

Basic helix-loop-helix (bHLH) transcription factors (TFs) are widely distributed in eukaryotes and play an important role in biological growth and development. The identification and functional analyses of bHLH genes/proteins in edible mushrooms (Agaricus bisporus) have yet to be reported. In the present study, we identified 10 putative bHLH members carrying the conserved bHLH domains. Phylogenetic analyses revealed that the 10 AbbHLHs were the closest to sequences of species belonging to 7 different fungal subgroups, which was supported by loop length, intron patterns, and key amino acid residues. The substantial increase after harvest and continuously elevated expression of AbbHLH1 during the development until the disruption of mushroom velum, and the preferential expression in cap and gill tissues suggest the important function of AbbHLH1 in postharvest development of A. bisporus. The relationship of arginine catabolism-related genes with the early stage of postharvest continuing development also was revealed by expression determination. Subcellular localization showed that AbbHLH1 could be localized in nucleus. Importantly, the electrophoretic mobility shift and dual-luciferase reporter assays showed that AbbHLH1 activated the promoters of AbOAT, AbSPDS, and AbSAMDC and suppressed the expression of AbARG, AbUREA, and AbODC, probably for the modulation of arginine catabolism and thus control of postharvest mushroom development. Taken together, the available data provide valuable functional insight into the role of AbbHLH proteins in postharvest mushrooms.

Consumers' Preferences and Attitudes towards Plant-Based Milk.

Plant-based milk (PBM) has become increasingly popular due to its environmental sustainability, health benefits, ingredient abundance, and unique taste. This study aims to identify the main factors that affect consumer preferences and attitudes towards PBM, and to examine the effect of consumer attitudes including environmental awareness, health consciousness, and food neophobia on WTP. We use the double-bounded dichotomy choice (DBDC) method to calculate consumers' willingness to pay (WTP) for PBM. We find that the appearance, taste, nutritional value, and environmental benefits of PBM significantly increase consumers' WTP for it. Consumers with high environmental awareness are more likely to perceive PBM as environmentally friendly and are willing to pay a higher price for it. Consumers with high health consciousness tend to value the environmental benefits of PBM and prioritize purchase convenience, as it aligns with their health-conscious lifestyle, leading to a higher WTP for PBM. The results of our study can help design effective strategies to market plant-based milk and develop sustainable and healthy food systems.

Understanding the Interaction between Regulatory Focus and Message Framing in Determining Chinese Consumers' Attitudes toward Artificial Meat.

While production and consumption of meat cast a shadow over the prospects for sustainable development, artificial meat may be the solution. However, consumer acceptability of artificial meat is a major impediment to its use as a suitable alternative. This study analyzed the relationship between regulatory focus and consumer acceptance of artificial meat using randomized controlled trial data. Results showed that promotion focus results in a higher acceptance of artificial meat products due to a higher perceived benefit and lower perceived risk, whereas prevention focus results in a lower acceptance of artificial meat products due to perceived benefit being lower and perceived risk being higher. The moderating effect of the message framing was investigated employing structural equation modeling (SEM). It was discovered that a gain-oriented message framing could greatly strengthen the association between promotion focus and perceived benefit, whereas an avoidance-oriented message framing could significantly diminish the relationship between prevention focus and perceived risk. This study has crucial implications for how policymakers and industries communicate with consumers about artificial meat.

Deep learning accurately predicts food categories and nutrients based on ingredient statements.

Determining attributes such as classification, creating taxonomies and nutrients for foods can be a challenging and resource-intensive task, albeit important for a better understanding of foods. In this study, a novel dataset, 134 k BFPD, was collected from USDA Branded Food Products Database with modification and labeled with three food taxonomy and nutrient values and became an artificial intelligence (AI) dataset that covered the largest food types to date. Overall, the Multi-Layer Perceptron (MLP)-TF-SE method obtained the highest learning efficiency for food natural language processing tasks using AI, which achieved up to 99% accuracy for food classification and 0.98 R2 for calcium estimation (0.93 âˆ¼ 0.97 for calories, protein, sodium, total carbohydrate, total lipids, etc.). The deep learning approach has great potential to be embedded in other food classification and regression tasks and as an extension to other applications in the food and nutrient scope.

Application of deep learning for image-based Chinese market food nutrients estimation.

With commercialization of deep learning (DL) models, daily precision dietary record based on images from smartphones becomes possible. This study took advantage of DL techniques on visual recognition tasks and proposed a suite of big-data-driven DL models regressing from food images to their nutrient estimation. We established and publicized the first food image database from the Chinese market, named ChinaMartFood-109. It contained 10,921 images with 23 nutrient contents, covering 18 main food groups. Inception V3 was optimized using other state-of-the-art deep convolutional neural networks, achieving up to 78 % and 94 % for top-1 and top-5 accuracy, respectively. Besides, this research compared three nutrient estimation algorithms and achieved the best regression coefficient (R2) by normalization + AM compared with arithmetic mean and harmonic mean, validating applicability in practice as well as theory. These encouraging results provide further evidence supporting artificial intelligence in the field of food analysis.

Image-based nutrient estimation for Chinese dishes using deep learning.

Food image recognition systems facilitate dietary assessment and in turn track users' dietary behaviors. However, due to the diversity of Chinese food, a quick and accurate food image recognizing is a particularly challenging task. The success of deep learning in computer vision inspired us to investigate its potential in this task. To satisfy its requirement on large-scale data, we established the first open-access image database for Chinese dishes, named ChinaFood-100, with quantitative nutrient annotations. We collected 10,074 images covering 100 food categories, including staple, meat, seafood, and vegetables. Based on this dataset, we trained four state-of-art deep learning neural network architectures for image recognition and showed that deep learning model Inception V3 resulted in the most advantageous recognition performance 78.26% in top-1 accuracy and 96.62% in top-5 accuracy. Based on this image recognition posterior, we further compared three nutrition estimation algorithms for food nutrient estimation. The results showed that the top-5 Arithmetic Mean (AM) algorithm achieved the highest regression coefficient (R2) up to 0.73 for protein estimation, which validated its applicability in practice. In addition, we analyzed our algorithm in terms of precision-recall and Grad-CAM. The results achieved by deep learning for food nutrient estimation may encourage artificial intelligence to be applied to the field of food, which shed the light on improvement in the future.

Over-expression of SlWRKY46 in tomato plants increases susceptibility to Botrytis cinerea by modulating ROS homeostasis and SA and JA signaling pathways.

WRKY, as one of the largest families of transcription factors (TFs), binds to cis-acting elements of downstream genes to regulate biotic and abiotic stress. However, the role of SlWRKY46 in fungal disease response induced by Botrytis cinerea (B.cinerea) and potential mechanism remains obscure. To ascertain the role of SlWRKY46 in response to B.cinerea, we constructed SlWRKY46-overexpression plants, which were then inoculated with B.cinerea. SlWRKY46-overexpression plants were more susceptible to B.cinerea and accompanied by the inhibited activities of phenylalanine ammonialyase (PAL), polyphenol oxidase (PPO), chitinase (CHI), and ß-1,3-glucanase (GLU). Additionally, SlWRKY46-overexpression plants showed the decreased activities of ascorbate peroxidase (APX), superoxide dismutase (SOD) and the content of H2O2, and the increased content of O2•-. Moreover, over-expression of SlWRKY46 suppressed the salicylic acid (SA) and jasmonic acid (JA) marker genes, pathogenesis related protein (PR1), and proteinase inhibitors (PI Ⅰ and PI Ⅱ) and consequently aggravated the disease symptoms. Therefore, we speculated that SlWRKY46 played negative regulatory roles in B. cinerea infection probably by inhibiting the activities of antioxidants and disease resistance enzymes, regulating SA and JA signaling pathways and modulating reactive oxygen (ROS) homeostasis.

Knockout of SlNPR1 enhances tomato plants resistance against Botrytis cinerea by modulating ROS homeostasis and JA/ET signaling pathways.

Tomato is one of the most popular horticultural crops, and many commercial tomato cultivars are particularly susceptible to Botrytis cinerea. Non-expressor of pathogenesis-related gene 1 (NPR1) is a critical component of the plant defense mechanisms. However, our understanding of how SlNPR1 influences disease resistance in tomato is still limited. In this study, two independent slnpr1 mutants were used to study the role of SlNPR1 in tomato resistance against B. cinerea. Compared to (WT), slnpr1 leaves exhibited enhanced resistance against B. cinerea with smaller lesion sizes, higher activities of chitinase (CHI), ß-1, 3-glucanases (GLU) and phenylalanine ammonia-lyase (PAL), and significantly increased expressions of pathogenesis-related genes (PRs). The increased activities of peroxidase (POD), ascorbate peroxidase (APX) and decreased catalase (CAT) activities collectively regulated reactive oxygen species (ROS) homeostasis in slnpr1 mutants. The integrity of the cell wall in slnpr1 mutants was maintained. Moreover, the enhanced resistance was further reflected by induction of defense genes involved in jasmonic acid (JA) and ethylene (ET) signaling pathways. Taken together, these findings revealed that knocking out SlNPR1 resulted in increased activities of defense enzymes, changes in ROS homeostasis and integrity of cell walls, and activation of JA and ET pathways, which confers resistance against B. cinerea in tomato plants.

Nitric Oxide Plays an Important Role in ß-Aminobutyric Acid-Induced Resistance to Botrytis cinerea in Tomato Plants.

ß-Aminobutyric acid (BABA) has consistently been reported to enhance plant immunity. However, the specific mechanisms and downstream components that mediate this resistance are not yet agreed upon. Nitric oxide (NO) is an important signal molecule involved in a diverse range of physiological processes, and whether NO is involved in BABA-induced resistance is interesting. In this study, treatment with BABA significantly increased NO accumulation and reduced the sensitivity to Botrytis cinerea in tomato plants. BABA treatment reduced physical signs of infection and increased both the transcription of key defense marker genes and the activity of defensive enzymes. Interestingly, compared to treatment with BABA alone, treatment with BABA plus cPTIO (NO specific scavenger) not only significantly reduced NO accumulation, but also increased disease incidence and lesion area. These results suggest that NO accumulation plays an important role in BABA-induced resistance against B. cinerea in tomato plants.

Knockout of SlMAPK3 enhances tolerance to heat stress involving ROS homeostasis in tomato plants.

BACKGROUND: High temperature is a major environmental stress that limits plant growth and agriculture productivity. Mitogen-activated protein kinases (MAPKs) are highly conserved serine and threonine protein kinases that participate in response to diverse environmental stresses in plants. A total of 16 putative SlMAPK genes are identified in tomato, and SlMAPK3 is one of the most extensively studied SlMAPKs. However, the role of SlMAPK3 in response to heat stress is not clearly understood in tomato plants. In this study, we performed functional analysis of SlMAPK3 for its possible role in response to heat stress. RESULTS: qRT-PCR analyses revealed that SlMAPK3 relative expression was depressed by heat stress. Here, wild-type (WT) tomato plants and CRISPR/Cas9-mediated slmapk3 mutant lines (L8 and L13) were used to investigate the function of SlMAPK3 in response to heat stress. Compared with WT plants, slmapk3 mutants exhibited less severe wilting and less membrane damage, showed lower reactive oxygen species (ROS) contents, and presented higher both activities and transcript levels of antioxidant enzymes, as well as elevated expressions of genes encoding heat stress transcription factors (HSFs) and heat shock proteins (HSPs). CONCLUSIONS: CRISPR/Cas9-mediated slmapk3 mutants exhibited more tolerance to heat stress than WT plants, suggesting that SlMAPK3 was a negative regulator of thermotolerance. Moreover, antioxidant enzymes and HSPs/HSFs genes expression were involved in SlMAPK3-mediated heat stress response in tomato plants.

Melatonin Induces Disease Resistance to Botrytis cinerea in Tomato Fruit by Activating Jasmonic Acid Signaling Pathway.

Melatonin acts as a crucial signaling molecule with multiple physiological functions in plant response to abiotic and biotic stresses. However, the impact and regulatory mechanism of melatonin on attenuating tomato fruit fungal decay are unclear. In this study, we investigated the potential roles of melatonin in modulating fruit resistance to Botrytis cinerea and explored related physiological and molecular mechanisms. The results revealed that disease resistance was strongly enhanced by melatonin treatment, and 50 µM was confirmed as the best concentration. Melatonin treatment increased the activities of defense-related enzymes and decreased hydrogen peroxide (H2O2) content with enhanced antioxidant enzyme activities. Moreover, we found that melatonin treatment increased methyl jasmonate (MeJA) content; up-regulated the expressions of SlLoxD, SlAOC, and SlPI II; and reduced the expressions of SlMYC2 and SlJAZ1. We postulated that melatonin played a positive role in tomato fruit resistance to Botrytis cinerea through regulating H2O2 level and JA signaling pathway.

Ethylene Perception Is Associated with Methyl-Jasmonate-Mediated Immune Response against Botrytis cinerea in Tomato Fruit.

Jasmonic acid (JA)- and ethylene-mediated signaling pathways are reported to have synergistic effects on inhibiting gray mold. The present study aimed to explain the role of ethylene perception in methyl jasmonate (MeJA)-mediated immune responses. Results showed that exogenous MeJA enhanced disease resistance, accompanied by the induction of endogenous JA biosynthesis and ethylene production, which led to the activation of the phenolic metabolism pathway. Blocking ethylene perception using 1-methylcyclopropene (1-MCP) either before or after MeJA treatment could differently weaken the disease responses induced by MeJA, including suppressing the induction of ethylene production and JA contents and reducing activities of lipoxygenase and allene oxide synthase compared to MeJA treatment alone. Consequently, MeJA-induced elevations in the total phenolic content and the activities of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, 4-coumarate:coenzyme A ligase, and peroxidase were impaired by 1-MCP. These results suggested that ethylene perception participated in MeJA-mediated immune responses in tomato fruit.