To aid valorisation of beer brewing by-products, more insight into their composition is essential. We have analysed the phenolic compound composition of four brewing by-products, namely barley rootlets, spent grain, hot trub, and cold trub. The main phenolics detected were hydroxycinnamoylagmatines and dimers thereof. Barley rootlets contained the highest hydroxycinnamoylagmatine content and cold trub the highest dimer content. Additionally, variations in (dimeric) hydroxycinnamoylagmatine composition and content were observed in fourteen barley rootlet samples. The most abundant compound in all rootlets was the glycosylated 4-O-7\'/3-8\'-linked heterodimer of coumaroylagmatine and feruloylagmatine, i.e. CouAgm-4-O-7\'/3-8\'-(4\'Hex)-DFerAgm. Structures of glycosylated and hydroxylated derivatives of coumaroylagmatine were elucidated by NMR spectroscopy after their purification from a rootlet extract. An MS-based decision tree was developed, which aids in identifying hydroxycinnamoylagmatine dimers in complex mixtures. In conclusion, this study shows that the diversity of phenolamides and (neo)lignanamides in barley-derived by-products is larger than previously reported.

Phenolamides are important secondary metabolites in plant species. They play important roles in plant defense responses against pathogens and insect herbivores, protection against UV irradiation and floral induction and development. However, the accumulation and variation in phenolamides content in diverse maize lines and the genes responsible for their biosynthesis remain largely unknown. Here, we combined genetic mapping, protein regulatory network and bioinformatics analysis to further enhance the understanding of maize phenolamides biosynthesis. Sixteen phenolamides were identified in multiple populations, and they were all significantly correlated with one or several of 19 phenotypic traits. By linkage mapping, 58, 58, 39 and 67 QTLs, with an average of 3.9, 3.6, 3.6 and 4.2 QTLs for each trait were mapped in BBE1, BBE2, ZYE1 and ZYE2, explaining 9.47%, 10.78%, 9.51% and 11.40% phenotypic variation for each QTL on average, respectively. By GWAS, 39 and 36 significant loci were detected in two different environments, 3.3 and 2.8 loci for each trait, explaining 10.00% and 9.97% phenotypic variation for each locus on average, respectively. Totally, 58 unique candidate genes were identified, 31% of them encoding enzymes involved in amine and derivative metabolic processes. Gene Ontology term analysis of the 358 protein-protein interrelated genes revealed significant enrichment in terms relating to cellular nitrogen metabolism, amine metabolism. GRMZM2G066142, GRMZM2G066049, GRMZM2G165390 and GRMZM2G159587 were further validated involvement in phenolamides biosynthesis. Our results provide insights into the genetic basis of phenolamides biosynthesis in maize kernels, understanding phenolamides biosynthesis and its nutritional content and ability to withstand biotic and abiotic stress.

BACKGROUND: Goji berry is a general term for various plant species in the genus Lycium. Goji has long been historically used in traditional Chinese medicines. Goji is a representative tonic medicine that has the effects of nourishing the liver and kidney and benefiting the essence and eyesight. It has been widely used in the treatment of various diseases, including tinnitus, impotence, spermatorrhea and blood deficiency, since ancient times.
OBJECTIVE: This study aims to comprehensively summarize the quality evaluation methods of the main compounds in goji, as well as the current research status of the phenolamides in goji and their pharmacological effects, to explore the feasibility of using phenolamides as quality control markers and thus improve the quality and efficacy in goji.
METHODS: Relevant literature from PubMed, Web of Science, Science Direct, CNKI and other databases was comprehensively collected, screened and summarized.
RESULTS: According to the collected literature, the quality evaluation markers of goji in the Pharmacopoeia of the People\'s Republic of China are Lycium barbarum polysaccharide (LBP) and betaine. As a result of its structure complexity, only the total level of LBP can be determined, while betaine is not prominent in the pharmacological action of goji and lacks species distinctiveness. Neither of them can well explain the quality of goji. KuA and KuB are commonly used as quality evaluation markers of the Lycii cortex because of their high levels and suitable pharmacological activity. Goji is rich in polyphenols, carotenoids and alkaloids. Many studies have used the above compounds to establish quality evaluation methods but the results have not been satisfactory. Phenolamides have often been neglected in previous studies because of their low single compound levels and high separation difficulty. However, in recent years, the favorable pharmacological activities of phenolamides have been gradually recognized, and studies on goji phenolamides are greatly increasing. In addition, phenolamides have higher species distinctiveness than other compounds and can be combined with other compounds to better evaluate the quality of goji to improve its average quality.
CONCLUSIONS: The phenolamides in the goji are rich and play a key role in antioxidation, anti-inflammation, neuroprotection and immunomodulation. As a result of their characteristics, it is suitable to evaluate the quality by quantitative analysis of multi-components by single-marker and fingerprint. This method can be combined with other techniques to improve the quality evaluation system of goji, which lays a foundation for their effectiveness and provides a reference for new quality evaluation methods of similar herbal medicines.

Among plant metabolites, phenolamides, which are conjugates of hydroxycinnamic acid derivatives and polyamines, play important roles in plant adaptation to abiotic and biotic stresses. However, the molecular mechanisms underlying phenolamide metabolism and regulation as well as the effects of domestication and breeding on phenolamide diversity in tomato remain largely unclear. In this study, we performed a metabolite-based genome-wide association study and identified two biosynthetic gene clusters (BGC7 and BGC11) containing 12 genes involved in phenolamide metabolism, including four biosynthesis genes (two 4CL genes, one C3H gene, and one CPA gene), seven decoration genes (five AT genes and two UGT genes), and one transport protein gene (DTX29). Using gene co-expression network analysis we further discovered that SlMYB13 positively regulates the expression of two gene clusters, thereby promoting phenolamide accumulation. Genetic and physiological analyses showed that BGC7, BGC11 and SlMYB13 enhance drought tolerance by enhancing scavenging of reactive oxygen species and increasing abscisic acid content in tomato. Natural variation analysis suggested that BGC7, BGC11 and SlMYB13 were negatively selected during tomato domestication and improvement, leading to reduced phenolamide content and drought tolerance of cultivated tomato. Collectively, our study discovers a key mechanism of phenolamide biosynthesis and regulation in tomato and reveals that crop domestication and improvement shapes metabolic diversity to affect plant environmental adaptation.

Bee pollen is hailed as a treasure trove of human nutrition and has progressively emerged as the source of functional food and medicine. This review conducts a compilation of nutrients and phytochemicals in bee pollen, with particular emphasis on some ubiquitous and unique phenolamides and flavonoid glycosides. Additionally, it provides a concise overview of the diverse health benefits and therapeutic properties of bee pollen, particularly anti-prostatitis and anti-tyrosinase effects. Furthermore, based on the distinctive structural characteristics of pollen walls, a substantial debate has persisted in the past concerning the necessity of wall-disruption. This review provides a comprehensive survey on the necessity of wall-disruption, the impact of wall-disruption on the release and digestion of nutrients, and wall-disruption techniques in industrial production. Wall-disruption appears effective in releasing and digesting nutrients and exploiting bee pollen\'s bioactivities. Finally, the review underscores the need for future studies to elucidate the mechanisms of beneficial effects. This paper will likely help us gain better insight into bee pollen to develop further functional foods, personalized nutraceuticals, cosmetics products, and medicine.

OBJECTIVE: Diets and parasites influence the gut bacterial symbionts of bumble bees, but potential interactive effects remain overlooked. The main objective of this study was to assess the isolated and interactive effects of sunflower pollen, its phenolamides, and the widespread trypanosomatid Crithidia sp. on the gut bacterial symbionts of Bombus terrestris males.
RESULTS: Bumble bee males emerged in microcolonies fed on either (i) willow pollen (control), (ii) sunflower pollen, or (iii) willow pollen spiked with phenolamide extracts from sunflower pollen. These microcolonies were infected by Crithidia sp. or were pathogen-free. Using 16S rRNA amplicon sequencing (V3-V4 region), we observed a significant alteration of the beta diversity but not of the alpha diversity in the gut microbial communities of males fed on sunflower pollen compared to males fed on control pollen. Similarly, infection by the gut parasite Crithidia sp. altered the beta diversity but not the alpha diversity in the gut microbial communities of males, irrespective of the diet. By contrast, we did not observe any significant alteration of the beta or alpha diversity in the gut microbial communities of males fed on phenolamide-enriched pollen compared to males fed on control pollen. Changes in the beta diversity indicate significant dissimilarities of the bacterial taxa between the treatment groups, while the lack of difference in alpha diversity demonstrates no significant changes within each treatment group.
CONCLUSIONS: Bumble bees harbour consistent gut microbiota worldwide, but our results suggest that the gut bacterial communities of bumble bees are somewhat shaped by their diets and gut parasites as well as by the interaction of these two factors. This study confirms that bumble bees are suitable biological surrogates to assess the effect of diet and parasite infections on gut microbial communities.

Bee pollen (BP) and bee bread (BB) are natural food sources containing a wide variety of bioactive compounds, complementing their rich nutritional composition. These bee products are being explored to empower functional foods, with the term functionality being dependent on the bioactive compounds added to the food matrix. However, there is not enough evidence of the effect of heat on these compounds during food processing and production and how it impacts their biological activity. Here, we enriched traditional bread by adding BP and BB at different proportions of 1 to 5% and tested the thermal stability of their bioactive compounds through several spectroscopic and chromatographic analyses. Adding bee pollen and bee bread to bread resulted in a 4 and 5-fold increase in total phenolic content, respectively. While not all the 38 phenolic and phenolamide compounds identified in the raw BP and BB were detected in the processed bread, phenolamides were found to be more resilient to baking and heat treatment than flavonoids. Still, the enriched bread\'s antioxidant activity improved with the addition of BP and BB. Therefore, incorporating bee products into heat-treated products could enhance the functionality of staple foods and increase the accessibility to these natural products.

This study aimed at investigating phenolamides and flavonoid glycosides in 20 types of monofloral bee pollen. The plant origins of pollen samples were determined by DNA barcoding, with the purities to over 70 %. The 31 phenolamides and their 33 cis/trans isomers, and 25 flavonoid glycosides were identified; moreover, 19 phenolamides and 14 flavonoid glycosides as new-found compounds in bee pollen. All phenolics and flavonoids are present in the amidation or glycosylation form. The MS/MS cleavage modes of phenolamides and flavonoid glycosides were summarized. Isorhamnetin-3-O-gentiobioside presented the highest levels 23.61 mg/g in apricot pollen. Phenolamides in 11 types of pollen constituted over 1 % of the total weight, especially 3.9 % in rose and 2.8 % in pear pollen. Tri-p-coumaroyl spermidine and di-p-coumaroyl-caffeoyl spermidine respectively accounted for over 2.6 % of the total weight in pear and rose pollen. The richness in phenolamides and flavonoid glycosides can offer bee pollen more bioactivities as functional foods.

Phenolamide (PA) metabolites play important roles in the interaction between plants and pathogens. The putrescine hydroxycinnamoyl transferase genes OsPHT3 and OsPHT4 positively regulate rice cell death and resistance to Magnaporthe oryzae. The bZIP transcription factor APIP5, a negative regulator of cell death and rice immunity, directly binds to the OsPHT4 promoter to regulate putrescine-derived PAs. Whether other hydroxycinnamoyl transferase (HT) genes also participate in APIP5-mediated immunity remains unclear. Surprisingly, we find that genes encoding agmatine hydroxycinnamoyl transferases OsAHT1 and OsAHT2, tryptamine hydroxycinnamoyl transferases OsTBT1 and OsTBT2, and tyramine hydroxycinnamoyl transferases OsTHT1 and OsTHT2, responsible for the biosynthesis of polyamine-derived PAs are all up-regulated in APIP5-RNAi transgenic plants compared with segregated wild-type rice. Furthermore, both OsAHT1/2 and OsTBT1/2 are induced during M. oryzae infection, showing expression patterns similar to those previously reported for OsTHT1/2 and OsPHT3/4. Transgenic plants overexpressing either OsAHT2-GFP or OsTBT1-GFP show enhanced resistance against M. oryzae and accumulated more PA metabolites and lignin compared with wild-type plants. Interestingly, as demonstrated for OsPHT4, APIP5 directly binds to the promoters of OsAHT1/2, OsTBT1/2, and OsTHT1/2, repressing their transcription. Together, these results indicate that the HT genes are common targets of APIP5 and that PAs play critical roles in rice immunity.

Microbial fermentation can break the bee pollen wall. However, the global profiling of bee pollen metabolites under fermentation remains unclear. This study aims to comprehensively elucidate the changes in the composition of bee pollen after microbial fermentation. Ultra-performance liquid chromatography-electron spray ionization-mass spectrometry (UPLC-ESI-MS) based on widely targeted metabolomics analysis was used to compare the chemical composition of unfermented bee pollen (UBP) and fermented bee pollen (FBP). Among the 890 metabolites detected, a total of 668 differential metabolites (classified into 17 categories) were identified between UBP and FBP. Fermentation significantly increased the contents of primary metabolites such as 74 amino acids and derivatives, 42 polyunsaturated fatty acids and 66 organic acids, as well as some secondary metabolites such as 38 phenolic acids, 80 flavone aglycones and 22 phenolamides. The results indicate that fermentation is a promising strategy to improve the nutritional value of bee pollen.