Caffeoylquinic acid profiling: comparative analysis in yerba mate, Indian camphorweed, and stevia extracts with emphasis on the influence of brewing conditions and cold storage in yerba mate infusion.

Herbal infusions exhibit diverse pharmacological effects, such as antioxidant, anti-inflammatory, anticancer, antihypertensive, and antineurodegenerative activities, which can be attributed to the high content of phenolic compounds (e.g., caffeoylquinic acids (CQAs)). In this study, we used ultraperformance liquid chromatography to determine the content of CQAs in the methanolic extracts of model herbs, namely, yerba mate (Ilex paraguariensis), stevia (Stevia rebaudiana), and Indian camphorweed (Pluchea indica (L.) Less.). The results revealed that yerba mate had the highest total CQA content (108.05 ± 1.12 mg/g of dry weight). Furthermore, we evaluated the effect of brewing conditions and storage at 4 °C under dark and light conditions on the antioxidant property and total phenolic and CQA contents of a yerba mate infusion. The analysis of the yerba mate infusions prepared with different steeping times, dried leaf weights, and water temperatures revealed that the amount of extracted CQAs was maximized (∼175 mg/150 mL) when 6 g of dried leaves were steeped in hot water for 10 min. A total of 10-day refrigerated storage resulted in no significant changes in the antioxidant activity and total phenolic and CQA contents of an infusion kept in a brown container (dark). However, the antioxidant properties and total phenolic and CQA contents were negatively affected when kept in a clear container, suggesting the detrimental effect of light exposure. Our study provides practical recommendations for improving the preparation and storage of herbal infusions, thus catering to the needs of consumers, food scientists, and commercial producers. Moreover, it is the first study of the influence of light exposure on the content of crucial quality attributes within plant-based beverages.

Metabolomics for Agricultural Waste Valorization: Shifting Toward a Sustainable Bioeconomy.

Agriculture has been considered as a fundamental industry for human survival since ancient times. Local and traditional agriculture are based on circular sustainability models, which produce practically no waste. However, owing to population growth and current market demands, modern agriculture is based on linear and large-scale production systems, generating tons of organic agricultural waste (OAW), such as rejected or inedible plant tissues (shells, peels, stalks, etc.). Generally, this waste accumulates in landfills and creates negative environmental impacts. The plant kingdom is rich in metabolic diversity, harboring over 200,000 structurally distinct metabolites that are naturally present in plants. Hence, OAW is considered to be a rich source of bioactive compounds, including phenolic compounds and secondary metabolites that exert a wide range of health benefits. Accordingly, OAW can be used as extraction material for the discovery and recovery of novel functional compounds that can be reinserted into the production system. This approach would alleviate the undesired environmental impacts of OAW accumulation in landfills, while providing added value to food, pharmaceutical, cosmetic, and nutraceutical products and introducing a circular economic model in the modern agricultural industry. In this regard, metabolomics-based approaches have gained increasing interest in the agri-food sector for a variety of applications, including the rediscovery of bioactive compounds, owing to advances in analytical instrumentation and data analytics platforms. This mini review summarizes the major aspects regarding the identification of novel bioactive compounds from agricultural waste, focusing on metabolomics as the main tool.

Sprout Caffeoylquinic Acid Profiles as Affected by Variety, Cooking, and Storage.

Various health-promoting properties inherent to plant-based foods have been attributed to their rich bioactive compounds, including caffeoylquinic acids (CQAs). The potential health benefits of CQAs have been well-documented. While sprouts are widely recognized as health-promoting foods owing to their high phytonutrient content, our knowledge regarding the effect of cooking and storage, commonly practiced by consumers, on the CQA content remains limited. First, sunflower sprouts were found to have the highest total CQA content (~ 22 mg/g dry weight) out of 11 commonly available sprouts. Then, the effect of variety, cooking, and low-temperature storage on the CQA profile of sunflower sprouts was investigated. Among the four different varieties of sunflower sprouts, variety 1 harbored the highest total CQA content. Notably, cooking adversely affected the CQA content of sunflower sprouts relative to the uncooked samples in a time-dependent manner, possibly due to the heat sensitivity of CQAs. Under simulated home-refrigeration storage conditions, we observed a significant decline in the content of major CQA compounds (5-monoCQA and 3,5-diCQA) at days 10 and 13 of storage. The results obtained herein provide consumers and food industrialists with increased insight into the effect of cooking and refrigeration on the CQA content of sunflower sprouts.

Comprehensive genome-wide analysis of calmodulin-binding transcription activator (CAMTA) in Durio zibethinus and identification of fruit ripening-associated DzCAMTAs.

BACKGROUND: Fruit ripening is an intricate developmental process driven by a highly coordinated action of complex hormonal networks. Ethylene is considered as the main phytohormone that regulates the ripening of climacteric fruits. Concomitantly, several ethylene-responsive transcription factors (TFs) are pivotal components of the regulatory network underlying fruit ripening. Calmodulin-binding transcription activator (CAMTA) is one such ethylene-induced TF implicated in various stress and plant developmental processes. RESULTS: Our comprehensive analysis of the CAMTA gene family in Durio zibethinus (durian, Dz) identified 10 CAMTAs with conserved domains. Phylogenetic analysis of DzCAMTAs, positioned DzCAMTA3 with its tomato ortholog that has already been validated for its role in the fruit ripening process through ethylene-mediated signaling. Furthermore, the transcriptome-wide analysis revealed DzCAMTA3 and DzCAMTA8 as the highest expressing durian CAMTA genes. These two DzCAMTAs possessed a distinct ripening-associated expression pattern during post-harvest ripening in Monthong, a durian cultivar native to Thailand. The expression profiling of DzCAMTA3 and DzCAMTA8 under natural ripening conditions and ethylene-induced/delayed ripening conditions substantiated their roles as ethylene-induced transcriptional activators of ripening. Similarly, auxin-suppressed expression of DzCAMTA3 and DzCAMTA8 confirmed their responsiveness to exogenous auxin treatment in a time-dependent manner. Accordingly, we propose that DzCAMTA3 and DzCAMTA8 synergistically crosstalk with ethylene during durian fruit ripening. In contrast, DzCAMTA3 and DzCAMTA8 antagonistically with auxin could affect the post-harvest ripening process in durian. Furthermore, DzCAMTA3 and DzCAMTA8 interacting genes contain significant CAMTA recognition motifs and regulated several pivotal fruit-ripening-associated pathways. CONCLUSION: Taken together, the present study contributes to an in-depth understanding of the structure and probable function of CAMTA genes in the post-harvest ripening of durian.

Transcriptome-wide identification and expression profiling of the ERF gene family suggest roles as transcriptional activators and repressors of fruit ripening in durian.

The involvement of the phytohormone ethylene as the main trigger of climacteric fruit ripening is well documented. However, our knowledge regarding the role of ethylene response factor (ERF) transcription factor in the transcriptional regulation of ethylene biosynthesis during fruit ripening remains limited. Here, comprehensive transcriptome analysis and expression profiling revealed 63 ERFs in durian pulps, termed DzERF1-DzERF63, of which 34 exhibited ripening-associated expression patterns at three stages (unripe, midripe, and ripe) during fruit ripening. Hierarchical clustering analysis classified 34 ripening-associated DzERFs into three distinct clades, among which, clade I consisted of downregulated DzERFs and clade III included those upregulated during ripening. Phylogenetic analysis predicted the functions of some DzERFs based on orthologs of previously characterized ERFs. Among downregulated DzERFs, DzERF6 functional prediction revealed its role as a negative regulator of ripening via ethylene biosynthetic gene repression, whereas among upregulated genes, DzERF9 was predicted to positively regulate ethylene biosynthesis. Correlation network analysis of 34 ripening-associated DzERFs with potential target genes revealed a strong negative correlation between DzERF6 and ethylene biosynthetic genes and a strong positive correlation between DzERF9 and ethylene biosynthesis. DzERF6 and DzERF9 showed differential expression patterns in association with different ripening treatments (natural, ethylene-induced, and 1-methylcyclopropene-delayed ripening). DzERF6 was downregulated, whereas DzERF9 was upregulated, during ripening and after ethylene treatment. The auxin-repressed and auxin-induced expression of DzERF6 and DzERF9, respectively, confirmed its dose-dependent responsiveness to exogenous auxin. We suggest ethylene- and auxin-mediated roles of DzERF6 and DzERF9 during fruit ripening, possibly through transcriptional regulation of ethylene biosynthetic genes.

Assessing Dynamic Changes of Taste-Related Primary Metabolism During Ripening of Durian Pulp Using Metabolomic and Transcriptomic Analyses.

Durian is an economically important fruit of Southeast Asia. There is, however, a lack of in-depth information on the alteration of its metabolic networks during ripening. Here, we annotated 94 ripening-associated metabolites from the pulp of durian cv. Monthong fruit at unripe and ripe stages, using capillary electrophoresis- and gas chromatography- time-of-flight mass spectrometry, specifically focusing on taste-related metabolites. During ripening, sucrose content increased. Change in raffinose-family oligosaccharides are reported herein for the first time. The malate and succinate contents increased, while those of citrate, an abundant organic acid, were unchanged. Notably, most amino acids increased, including isoleucine, leucine, and valine, whereas aspartate decreased, and glutamate was unchanged. Furthermore, transcriptomic analysis was performed to analyze the dynamic changes in sugar metabolism, glycolysis, TCA cycle, and amino acid pathways to identify key candidate genes. Taken together, our results elucidate the fundamental taste-related metabolism of durian, which can be exploited to develop durian metabolic and genetic markers in the future.

Auxin Response Factor 2A Is Part of the Regulatory Network Mediating Fruit Ripening Through Auxin-Ethylene Crosstalk in Durian.

Fruit ripening is a highly coordinated developmental process driven by a complex hormonal network. Ethylene is the main regulator of climacteric fruit ripening. However, a putative role of other key phytohormones in this process cannot be excluded. We previously observed an increasing level of auxin during the post-harvest ripening of the durian fruit, which occurred concomitantly with the rise in the climacteric ethylene biosynthesis. Herein, we connect the key auxin signaling component, auxin response factors (ARFs), with the regulatory network that controls fruit ripening in durian through the identification and functional characterization of a candidate ripening-associated ARF. Our transcriptome-wide analysis identified 15 ARF members in durian (DzARFs), out of which 12 were expressed in the fruit pulp. Most of these DzARFs showed a differential expression, but DzARF2A had a marked ripening-associated expression pattern during post-harvest ripening in Monthong, a commercial durian cultivar from Thailand. Phylogenetic analysis of DzARF2A based on its tomato orthologue predicted a role in ripening through the regulation of ethylene biosynthesis. Transient expression of DzARF2A in Nicotiana benthamiana leaves significantly upregulated the expression levels of ethylene biosynthetic genes, pointing to a ripening-associated role of DzARF2A through the transcriptional regulation of ethylene biosynthesis. Dual-luciferase reporter assay determined that DzARF2A trans-activates durian ethylene biosynthetic genes. We previously reported significantly higher auxin level during post-harvest ripening in a fast-ripening cultivar (Chanee) compared to a slow-ripening one (Monthong). DzARF2A expression was significantly higher during post-harvest ripening in the fast-ripening cultivars (Chanee and Phuangmanee) compared to that of the slow-ripening ones (Monthong and Kanyao). Thus, higher auxin level could upregulate the expression of DzARF2A during ripening of a fast-ripening cultivar. The auxin-induced expression of DzARF2A confirmed its responsiveness to exogenous auxin treatment in a dose-dependent manner, suggesting an auxin-mediated role of DzARF2A in fruit ripening. We suggest that high DzARF2A expression would activate ARF2A-mediated transcription of ethylene biosynthetic genes, leading to increased climacteric ethylene biosynthesis (auxin-ethylene crosstalk) and faster ripening. Hence, we demonstrated DzARF2A as a new component of the regulatory network possibly mediating durian fruit ripening through transcriptional regulation of ethylene biosynthetic genes.

Genome-wide analysis of the Dof gene family in durian reveals fruit ripening-associated and cultivar-dependent Dof transcription factors.

DNA binding with one finger (Dof) proteins constitute a ubiquitous plant-specific transcription factor (TF) family associated with diverse biological processes, including ripening. We conducted a genome-wide analysis of durian (Durio zibethinus Murr.) and identified 24 durian Dofs (DzDofs), 15 of which were expressed in fruit pulp. Gene expression analysis revealed differential expression of DzDofs during ripening in two commercial durian cultivars from Thailand, Monthong and Chanee. Comparing the expression levels of fruit pulp-expressed DzDofs between cultivars revealed ten potential cultivar-dependent Dofs, among which DzDof2.2 showed a significantly greater fold increase at every ripening stage in Chanee than in Monthong. The prediction of DzDof2.2's function based on its orthologue in Arabidopsis revealed its possible role in regulating auxin biosynthesis. We observed significantly higher auxin levels during ripening of Chanee than Monthong which concurred with the greater expression of auxin biosynthetic genes. Transient expression of DzDof2.2 in Nicotiana benthamiana significantly upregulated the expression levels of auxin biosynthetic genes. Higher expression levels of DzDof2.2 in Chanee would enhance auxin levels through transcriptional regulation of auxin biosynthetic genes. Higher auxin levels in Chanee could activate auxin-mediated transcription, contributing to its faster ripening compared to Monthong through earlier initiation of the ethylene response (auxin-ethylene crosstalk).

Identification and Functional Characterization of Genes Involved in the Biosynthesis of Caffeoylquinic Acids in Sunflower (Helianthus annuus L.).

Sunflower (Helianthus annuus L.) sprouts accumulate high amounts of caffeoylquinic acids (CQAs) including chlorogenic acid (5-CQA) and 1,5-diCQA. These compounds, which can be found in many plants, including tomato, globe artichoke, and chicory, have many health benefits, including antioxidant, antihepatotoxic, and antiglycative activities. However, CQA profiles and biosynthesis have not previously been studied in sunflower sprouts. In the present study, we found that 5-CQA and 1,5-diCQA were the major CQAs found in sunflower sprouts. We also identified minor accumulation of other CQAs, namely 3-CQA, 4-CQA, 3,4-diCQA, and 4,5-diCQA. According to genome-wide identification and phylogenetic analysis of genes involved in CQA biosynthesis in sunflower, three genes (HaHQT1, HaHQT2, and HaHQT3) encoding hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (HQT) and two genes (HaHCT1 and HaHCT2) encoding hydroxycinnamoyl CoA:shikimate/quinate hydroxycinnamoyl transferase (HCT) were identified. Expression analysis of these five genes in hypocotyls and cotyledons strongly suggested that HaHQT2 could be the main enzyme responsible for CQA biosynthesis, as HaHQT2 had the highest expression levels. In addition, when transiently expressed in the leaves of Nicotiana benthamiana, all three HaHQTs, which were soluble and not membrane-bound enzymes, could increase the content of 5-CQA by up to 94% compared to that in a control. Overall, our results increase understanding of CQA biosynthesis in sunflower sprouts and could be exploited by plant breeders to enhance accumulation of health-promoting CQAs in these plants.

Effect of cold-pressed and normal centrifugal juicing on quality attributes of fresh juices: do cold-pressed juices harbor a superior nutritional quality and antioxidant capacity?

Cold-pressed juices are claimed to contain higher levels of antioxidants and bioactive compounds compared to normally centrifuged ones. Herein, we evaluated the antioxidant capacity and the bioactive compound contents of some freshly prepared fruit juices, extracted by a cold-pressed juicer and compared them to those prepared by a normal centrifugal juicer. We observed no significant differences between cold-pressed and normal centrifugal juices in terms of the contents of bioactive compounds (ascorbic acid, total phenolic, and total carotenoid) and antioxidant capacity (ferric ion reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity). Storage at room temperature (∼28 °C) adversely affected the ascorbic acid, total phenolics, total carotenoids, FRAP and DPPH values of the cold-pressed juices within 48 h. However, under simulated home-refrigerated storage conditions, the antioxidant capacity, contents of bioactive compounds and physicochemical properties of the cold-pressed juices remained unchanged till day 5 post-storage. However, at day 6, most of the parameters exhibited a decreasing trend and reached their lowest values at day 7. Principal component analysis confirmed significant changes in the quality of juices at day 7 of storage related to the first two principal components (ascorbic acid and FRAP). Our results strongly question the claim regarding the superior quality of cold-pressed juices. Moreover, our findings provided compelling evidence regarding the possible adverse effects of long storage under home-refrigerated conditions on the quality of cold-pressed juices.

Effect of microorganisms on reducing cadmium uptake and toxicity in rice (Oryza sativa L.).

This study analyzed the application of three microorganism inoculums, including Bacillus subtilis, Bacillus cereus, and commercial effective microorganism (EM) solution in order to determine cadmium (Cd) reduction in rice (Oryza sativa L.) and rice growth promotion. Rice was grown in Cd-contaminated soil (120 mg/kg) and selected microorganisms were inoculated. Cd concentration and rice weight were measured at 45 and 120 days of the experiment. The result showed that B. subtilis inoculation into rice can highly reduce Cd accumulation in every part of rice roots and shoots (45 days), and grains (120 days). This species can effectively absorb Cd compared to other inoculums, which might be the main mechanism to reduce Cd transportation in rice plants. Interestingly, plants that were inoculated with bacterial species individually harbored higher calcium (Ca) and magnesium (Mg) accumulation; B. subtilis-inoculated plants had the highest levels of Ca and Mg compared to other inoculated ones. Moreover, inoculating rice plants with these microorganisms could increase the dry weight of the plant and protect them from Cd stress because all the inoculums presented the ability to solubilize phosphate, produce indole-3-acetic acid (IAA), and control ethylene levels by 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. After 120 days, quantification of each inoculum by quantitative polymerase chain reaction (qPCR) confirmed the root colonization of bacterial species, with B. subtilis showing higher 16S rRNA gene copy numbers than the other species. B. subtilis was classified as a non-human pathogenic strain, reassuring the safe application of this plant growth-promoting bacterium as a crop inoculum.

Impact of endophytic colonization patterns on Zamioculcas zamiifolia stress response and in regulating ROS, tryptophan and IAA levels under airborne formaldehyde and formaldehyde-contaminated soil conditions.

Deeper understanding of plant-endophyte interactions under abiotic stress would provide new insights into phytoprotection and phytoremediation enhancement. Many studies have investigated the positive role of plant-endophyte interactions in providing protection to the plant against pollutant stress through auxin (indole-3-acetic acid (IAA)) production. However, little is known about the impact of endophytic colonization patterns on plant stress response in relation to reactive oxygen species (ROS) and IAA levels. Moreover, the possible effect of pollutant phase on plant stress response is poorly understood. Here, we elucidated the impact of endophytic colonization patterns on plant stress response under airborne formaldehyde compared to formaldehyde-contaminated soil. ROS, tryptophan and IAA levels in the roots and shoots of endophyte-inoculated and non-inoculated plants in the presence and absence of formaldehyde were measured. Strain-specific quantitative polymerase chain reaction (qPCR) was used to investigate dynamics of endophyte colonization. Under the initial exposure to airborne formaldehyde, non-inoculated plants accumulated more tryptophan in the shoots (compared to the roots) to synthesize IAA. However, endophyte-inoculated plants behaved differently as they synthesized and accumulated more tryptophan in the roots and, hence, higher levels of IAA accumulation and exudation within roots which might act as a signaling molecule to selectively recruit B. cereus ERBP. Under continuous airborne formaldehyde stress, higher levels of ROS accumulation in the shoots pushed the plant to synthesize more tryptophan and IAA in the shoots (compared to the roots). Higher levels of IAA in the shoots might act as the potent driving force to relocalize B. cereus ERBP from roots to the shoots. In contrast, under formaldehyde-contaminated soil, B. cereus ERBP colonized root tissues without moving to the shoots since there was a sharp increase in ROS, tryptophan and IAA levels of the roots without any significant increase in the shoots. Pollutant phase affected endophytic colonization patterns and plant stress responses differently.

Euphorbia milii-native bacteria interactions under airborne formaldehyde stress: Effect of epiphyte and endophyte inoculation in relation to IAA, ethylene and ROS levels.

Better understanding of plant-bacteria interactions under stress is of the prime importance for enhancing airborne pollutant phytoremediation. No studies have investigated plant-epiphyte interactions compared to plant-endophyte interactions under airborne formaldehyde stress in terms of plant Indole-3-acetic acid (IAA), ethylene, reactive oxygen species (ROS) levels and pollutant removal efficiency. Euphorbia milii was inoculated with native plant growth-promoting (PGP) endophytic and epiphytic isolates individually to investigate plant-endophyte compared to plant-epiphyte interactions under continuous formaldehyde fumigation. Under airborne formaldehyde stress, endophyte interacts with its host plant closely and provides higher levels of IAA which protected the plant against formaldehyde phytotoxicity by lowering intracellular ROS, ethylene levels and maintaining shoot epiphytic community; hence, higher pollutant removal. However, plant-epiphyte interactions could not provide enough IAA to confer protection against formaldehyde stress; thus, increased ROS and ethylene levels, large decrease in shoot epiphytic population and lower pollutant removal although epiphyte contacts with airborne pollutant directly (has greater access to gaseous formaldehyde). Endophyte-inoculated plant synthesized more tryptophan as a signaling molecule for its associated bacteria to produce IAA compared to the epiphyte-inoculated one. Under stress, PGP endophyte interacts with its host closely; thus, better protection against stress and higher pollutant removal compared to epiphyte which has limited interactions with the host plant; hence, lower pollutant removal.

Euphorbia milii-Endophytic Bacteria Interactions Affect Hormonal Levels of the Native Host Differently Under Various Airborne Pollutants.

This study was conducted to assess the effect of plant-native endophytic bacteria interactions on indole-3-acetic acid (IAA), ethylene levels, and hormonal balance of Euphorbia milii under different airborne pollutants. IAA levels and airborne formaldehyde removal by E. milii enhanced when inoculated with endophytic isolates. However, one isolate, designated as root endophyte 4, with the highest levels of IAA production individually, declined gaseous formaldehyde removal of plant, since it disturbed hormonal balance of E. milii, leading to IAA levels higher than physiological concentrations, which stimulated ethylene biosynthesis and stomatal closure under light conditions. However, plant-root endophyte 4 interactions favored airborne benzene removal, since benzene was more phytotoxic and the plant needed more IAA to protect against benzene phytotoxicity. As trimethylamine (TMA) was not toxic, it did not affect plant-endophyte interactions. Therefore, IAA levels of root endophyte 4-inoculated E. milii was not significantly different from a noninoculated one. Under mixed-pollutant stress (formaldehyde, benzene, TMA), root endophyte 4-inoculated E. milii removed benzene at the lowest rate, since benzene was the most phytotoxic pollutant with the greatest molecular mass. However, TMA (with greater molecular mass) was removed faster than formaldehyde due to higher phytotoxicity of formaldehyde. Plant-endophyte interactions were affected differently under various airborne pollutants.

Effect of endophytic Bacillus cereus ERBP inoculation into non-native host: Potentials and challenges for airborne formaldehyde removal.

Phytoremediation could be a cost-effective, environmentally friendly approach for the treatment of indoor air. However, some drawbacks still dispute the expediency of phytotechnology. Our objectives were to investigate the competency of plant growth-promoting (PGP) endophytic Bacillus cereus ERBP (endophyte root blue pea), isolated from the root of Clitoria ternatea, to colonize and stabilize within Zamioculcas zamiifolia and Euphorbia milii as non-native hosts without causing any disease or stress symptoms. Moreover, the impact of B. cereus ERBP on the natural shoot endophytic community and for the airborne formaldehyde removal capability of non-native hosts was assessed. Non-native Z. zamiifolia was effectively inoculated with B. cereus ERBP through soil as the most efficient method of endophyte inoculation. Denaturing gradient gel electrophoresis profiling of the shoot endophytic community verified the colonization and stability of B. cereus ERBP within its non-native host during a 20-d fumigation period without interfering with the natural shoot endophytic diversity of Z. zamiifolia. B. cereus ERBP conferred full protection to its non-native host against formaldehyde phytotoxicity and enhanced airborne formaldehyde removal of Z. zamiifolia whereas non-inoculated plants suffered from formaldehyde phytotoxicity because their natural shoot endophytic community was detrimentally affected by formaldehyde. In contrast, B. cereus ERBP inoculation into non-native E. milii deteriorated airborne formaldehyde removal of the non-native host (compared to a non-inoculated one) as B. cereus ERBP interfered with natural shoot endophytic community of E. milii, which caused stress symptoms and stimulated ethylene biosynthesis. Non-native host inoculation with PGP B. cereus ERBP could bear potentials and challenges for airborne formaldehyde removal.