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Abstract Chinese wild rice (Zizania latifolia) is rich in phenolic compounds, particularly flavonoids. This study identified 203 basic helix‐loop‐helix (bHLHs) in Z. latifolia and showed that ZlbHLH196 (Zla16G011250) corresponds to the ZlRc gene in Z. latifolia, with its protein product localizing to the nucleus. Notably, the pericarps of ZlRc‐overexpressing (OE) rice are brown, whereas those of wild‐type (WT) rice are nonpigmented. The total phenolic, flavonoid, and proanthocyanidin contents, antioxidant activity, as well as enzyme inhibitory effects of ZlRc‐OE rice were significantly higher than those of WT rice. Overall, 221 differential phenolic metabolites were identified between ZlRc‐OE and WT rice, among which 198 were upregulated in the former. Additionally, a total of 227 differentially expressed genes were identified between ZlRc‐OE and WT rice, with 173 upregulated. Kyoto Encyclopedia of Genes and Genomes annotation and enrichment analysis of phenolic metabolites revealed enhanced isoflavonoid, flavone, flavonol, and flavonoid biosynthesis pathways in ZlRc‐OE rice, which, furthermore, showed a markedly upregulated expression and significantly higher activities of four key flavonoid biosynthesis–related enzymes (phenylalanine ammonia‐lyase, chalcone synthase, chalcone‐flavanone isomerase, and dihydroflavonol 4‐reductase). These findings show that ZlRc‐overexpression promotes phenolic compound accumulation in rice seeds and can be used to bioaugment rice phenolic content.

The rapid growth of the global population has led to an unprecedented demand for dietary protein. Canola seeds, being a widely utilized oil resource, generate substantial meal by-products following oil extraction. Fortunately, canola meals are rich in protein. In this present review, foremost attention is directed towards summarizing the characteristics of canola seed and canola seed protein. Afterwards, points of discussion related to pretreatment include an introduction to pulsed electric field treatment (PEF), microwave treatment (MC), and ultrasound treatment (UL). Then, the extraction method is illustrated, including alkaline extraction, isoelectric precipitation, acid precipitation, micellization (salt extraction), and dry fractionation and tribo-electrostatic separation. Finally, the structural complexity, physicochemical properties, and functional capabilities of rapeseed seeds, as well as the profound impact of various applications of rapeseed proteins, are elaborated. Through a narrative review of recent research findings, this paper aims to enhance a comprehensive understanding of the potential of canola seed protein as a valuable nutritional supplement, highlighting the pivotal role played by various extraction methods. Additionally, it sheds light on the broad spectrum of applications where canola protein demonstrates its versatility and indispensability as a resource.

Quinoa (Chenopodium quinoa Wild.) is a pseudo-grain that belongs to the amaranth family and has gained attention due to its exceptional nutritional properties. Compared to other grains, quinoa has a higher protein content, a more balanced amino acid profile, unique starch features, higher levels of dietary fiber, and a variety of phytochemicals. In this review, the physicochemical and functional properties of the major nutritional components in quinoa are summarized and compared to those of other grains. Our review also highlights the technological approaches used to improve the quality of quinoa-based products. The challenges of formulating quinoa into food products are addressed, and strategies for overcoming these challenges through technological innovation are discussed. This review also provides examples of common applications of quinoa seeds. Overall, the review underscores the potential benefits of incorporating quinoa into the diet and the importance of developing innovative approaches to enhance the nutritional quality and functionality of quinoa-based products.

Natural by-products, especially flavonoids, are in great demand in the nutra-pharmaceutical and biomedical industries. In this study, Ontario-grown onion varieties, namely Stanley, Safrane, Fortress, Lasalle and Ruby Ring, were screened for their antioxidant properties. Pressurized low polarity water technology, an environmentally friendly technique, was employed to extract the flavonoids from the onion varieties, followed by quantification and analysis using High Performance Liquid Chromatography. The antioxidant activities in the extracted samples were determined using various antioxidant assays, such as 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), ferric reducing ability of plasma, lipid peroxidation, total antioxidant capacity and oxygen radical absorbance capacity assays. The total phenolic content extracted from the Ruby Ring variety was the highest when compared to all the other yellow onion varieties tested. Our results indicate that Ruby Ring may be chosen as a preferred variety over other onion varieties to develop functional and health food products.

Wild soybean (Glycine soja Sieb.et Zucc; WS) has been used as a traditional food in China for many years and contains significantly higher levels of isoflavones than cultivated soybean (Glycine max; CS), but the secondary metabolites, including flavonoids and the phenolic composition differences between them, remain unclear. The results showed that WS possessed significantly higher total phenolic and flavonoid content and exhibited better antioxidant and α-glucosidase inhibition activities as well as excellent protective effects against H2O2-induced oxidative injury in a human endothelial cell line. Through metabolomic analysis, 642 metabolites were identified, and 238 showed differential expression, with 151 upregulated and 87 downregulated. A total of 79 flavonoid compounds were identified, 42 of which were upregulated in WS. 2′-Hydroxygenistein, garbanzol, protocatechuic aldehyde, ligustilide, and resveratrol were the most discriminated compounds in WS. The metabolic pathway analysis of differential metabolites related to the biosynthesis of flavonoids and phenolic acids were the biosynthesis of phenylpropanoids, flavonoids, isoflavonoids, flavones, and flavonols. This study substantially elucidated differences in the content of flavonoids and biological activities between WS and CS, which is useful information for the effective utilization of these two black soybean species in food processing.

Nicotiana tabacum solanesyl diphosphate synthase 1 (NtSPS1) is the key enzyme in solanesol biosynthesis. However, changes in the solanesol content, plant growth, photosynthesis, and metabolome of tobacco plants after NtSPS1 overexpression (OE) have not been previously reported. In the present study, these parameters, as well as photosynthetic gas exchange, chlorophyll content, and chlorophyll fluorescence parameters, were compared between NtSPS1 OE and wild type (WT) lines of tobacco. As expected, NtSPS1 OE significantly increased solanesol content in tobacco leaves. Although NtSPS1 OE significantly increased leaf growth, photosynthesis, and chlorophyll content, the chlorophyll fluorescence parameters in the leaves of the NtSPS1 OE lines were only slightly higher than those in the WT leaves. Furthermore, NtSPS1 OE resulted in 64 differential metabolites, including 30 up-regulated and 34 down-regulated metabolites, between the OE and WT leaves. Pathway enrichment analysis of these differential metabolites identified differentially enriched pathways between the OE and WT leaves, e.g., carbon fixation in photosynthetic organisms. The maximum carboxylation rate of RuBisCO and the maximum rate of RuBP regeneration were also elevated in the NtSPS1 OE line. To our knowledge, this is the first study to confirm the role of NtSPS1 in solanesol biosynthesis and its possible functional mechanisms in tobacco.

Longan fruit is a popular subtropical fruit with a relatively short shelf life at room temperature mainly due to pericarp browning and fungal infection. This study aimed to investigate the infection of Phomopsis longanae Chi in longan fruit and its effects on the storability and shelf life of longan fruit. The relationship between the energy metabolism of harvested longan fruit and disease development and pericarp browning was elucidated. Results show that P. longanae-inoculation accelerated the deterioration of longan fruit and caused pericarp browning. It also led to the energy deficit in pericarp of longan fruit, which was reflected as lower contents of ATP and ADP, higher AMP content, and lower energy charge as compared to the control samples. Additionally, P. longanae-infection reduced the activities of H+-ATPase, Ca2+-ATPase, and Mg2+-ATPase in plasma, vacuolar, and mitochondrial membranes during the storage period. The results demonstrate that P. longanae-infection led to disease development and pericarp browning in harvested longan fruit, which were due to the infection-induced energy deficit and low ATPase activity that caused disorders of ion transport and distribution, and damaged the structure and function of vacuole, mitochondria, and eventually the whole cells of fruit tissues.

The main goal of this study was to investigate the influences of Phomopsis longanae Chi infection on activities of cell wall-degrading enzymes (CWDEs), and contents of cell wall components in pericarp of harvested “Fuyan” longan (Dimocarpus longan Lour. cv. Fuyan) fruit and its relation to disease development. The results showed that, compared with the control samples, P. longanae-inoculated longans showed higher fruit disease index, lower content of pericarp cell wall materials (CWMs), as well as lower contents of pericarp cell wall components (chelate-soluble pectin (CSP), sodium carbonate-soluble pectin, hemicelluloses, and cellulose), but higher content of pericarp water-soluble pectin (WSP). In addition, the inoculation treatment with P. longanae significantly promoted the activities of CWDEs including pectinesterase, polygalacturonase, β-galactosidase, and cellulase. The results suggested that the P. longanae stimulated-disease development of harvested longans was due to increase in activities of pericarp CWDEs, which might accelerate the disassembly of pericarp cell wall components. In turn, resulting in the degradation of pericarp cell wall, reduction of pericarp mechanical strength, and subsequently leading to the breakdown of longan pericarp tissues. Eventually resulting in development of disease development and fruit decay in harvested longans during storage at 28°C.

The effects of fermentation conditions (temperature, time, and pH) and plucking standards (one leaf and a bud to four leaves and a bud) on the chemical components and sensory quality of the fermented juices processed from crushed fresh tea leaves were investigated. The results showed that optimum fermentation conditions that resulted in fermented juices of the best sensory quality and the highest content of TFs were a temperature of 35°C, time duration of 75 min, and pH 5.1. The fermented juices processed from new shoots with three leaves and a bud or four leaves and a bud afforded high overall acceptability and TF concentration. These differences arise because tea leaves with different plucking standards have different catechin content and enzyme activities. Fermented tea juice possessed higher concentrations of chemical components such as soluble solids, amino acids, and TFs and exhibited better sensory quality as compared to black tea infusion. The TF concentrations decreased as the pH of the fermenting juice increased, and the fermented juice showed the best overall acceptability. These results provide essential information for the improvement of the processing of black tea beverage by suggesting fermentation of fresh tea leaves as a better alternative to their infusion.

To explore Auricularia auricula-judae polysaccharides (AAP) as natural anticoagulants for application in the functional food industry, ultrasound assisted extraction (UAE) was optimized for the extraction of AAP by using a response surface methodology (RSM). The maximum extraction yield of crude AAP (14.74 mg/g) was obtained at the optimized extraction parameters as follows: Extraction temperature (74 °C), extraction time (27 min), the ratio of liquid to raw material (103 mL/g), and ultrasound power (198 W). Furthermore, the acidic AAP (aAAP) was precipitated with cetyltrimethylammonium bromide (CTAB) from crude AAP (cAAP). aAAP was further purified using ion exchange chromatography with a DEAE Purose 6 Fast Flow column to obtain aAAP-1. Additionally, according to the HPLC analysis, the aAAP-1 was mainly composed of mannose, glucuronic acid, glucose, galactose, and xylose, with a molar ratio of 80.63:9.88:2.25:1:31.13. Moreover, the results of the activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) indicated aAAP-1 had anticoagulant activity, which was a synergic anticoagulant activity by the endogenous and exogenous pathway.

The total polyphenol (TP), flavonoid, proanthocyanidin (PAC) content, and antioxidant capacity of both onion varieties (red and yellow) were compared. The content of TP, flavonoids, and PAC was determined by Folin-Ciocalteu colorimetric method, AlCl3, and by DMAC colorimetric method, respectively. The results showed that the contents of TP and flavonoids decreased from the outer to the inner layers in both onions, but there was no significant difference in PAC content. The outer layers had the highest antioxidant activity of extracts followed by a continuous decrease towards the inner layers in both varieties. The contents of phenolic acids and flavonoids were quantified by HPLC. Gallic acid, ferulic acid, and quercetin, as the main compounds in polyphenols, were detected in each layer of both onions. The red variety showed better antioxidant activity than yellow onion according to the linoleic acid system and DPPH assay. The higher contents of TP and flavonoids were associated with higher antioxidant activity.

Three polysaccharide-protein complexes of longan pulp (LP1-3) were isolated in this work. Their physicochemical characteristics and immunomodulatory effects on splenocytes, natural killer (NK) cells and macrophages in vitro were investigated. The carbohydrate portions of LP1-3 were principally composed of glucose, arabinose and mannose. LP3 displayed the maximal moisture absorption, and the thermal stability of LP2 was obviously higher than that of LP1 and LP3. All of them showed the characteristic polysaccharide and protein bands in the Fourier Transform Infrared (FTIR) spectrum. For a certain dose, all the fractions could significantly stimulate splenocyte proliferation, macrophage phagocytosis against neutral red, and NK cell cytotoxicity against YAC-1 lymphoma cell (P < 0.05). The results demonstrated that the polysaccharide-protein complexes of longan pulp have medical potential as immunotherapeutic adjuvants due to their immunomodulatory activities.

Castanopsis lamontii is traditionally used to prevent inflammatory diseases such as periodontitis and pharyngitis by residents in southwest China. However, little scientific evidence has been found to support this. In this research, the antibacterial activities of Castanopsis lamontii water extract (CLE) were assessed using the micro-dilution method. The anti-inflammatory and antioxidant activities of CLE were investigated in RAW264.7 cells. Key bioactive compounds in CLE were also explored. Results showed that CLE was capable of inhibiting the periodontitis pathogen Porphyromonas gingivalis and the pharyngitis pathogen β-hemolytic Streptococcus. It suppressed lipopolysaccharide-induced inflammation in RAW 264.7 cells via inactivating the TLR4/NF-κB pathway. Besides, it reduced oxidative stress-induced cell injury via scavenging reactive oxygen species. Chemical composition analysis revealed that CLE was rich in epicatechin and procyanidin B2. Further studies confirmed that epicatechin predominantly contributed to the antibacterial activities of CLE, while procyanidin B2 was mainly responsible for the anti-inflammatory activities of CLE. Both compounds contributed to the antioxidant activities of CLE. Acute oral toxicity tests proved that CLE was practically non-toxic. These results provide experimental evidences of the health-beneficial effects of CLE and may help promote the application of CLE in the food and health industries.

Solanesol is a terpene alcohol composed of nine isoprene units that mainly accumulates in solanaceous plants, especially tobacco (Nicotiana tabacum). The present study aimed to investigate the regulation of solanesol accumulation in tobacco leaves induced by moderately high temperature (MHT). Exposure to MHT resulted in a significant increase in solanesol content, dry weight, and net photosynthetic rate in tobacco leaves. In MHT-exposed tobacco leaves, 492 and 1440 genes were significantly up- and downregulated, respectively, as revealed by RNA-sequencing. Functional enrichment analysis revealed that most of the differentially expressed genes (DEGs) were mainly related to secondary metabolite biosynthesis, metabolic pathway, carbohydrate metabolism, lipid metabolism, hydrolase activity, catalytic activity, and oxidation-reduction process. Moreover, 122 transcription factors of DEGs were divided into 22 families. Significant upregulation of N. tabacum 3-hydroxy-3-methylglutaryl-CoA reductase (NtHMGR), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (NtDXR), geranylgeranyl diphosphate synthase (NtGGPS), and solanesyl diphosphate synthase (NtSPS) and significant downregulation of N. tabacum 1-deoxy-d-xylulose 5-phosphate synthase (NtDXS) and farnesyl diphosphate synthase (NtFPS) transcription upon MHT exposure were monitored by quantitative real-time PCR (qRT-PCR). This study indicated that solanesol accumulation in tobacco leaves can be manipulated through regulation of the environmental temperature and established a basis for further elucidation of the molecular mechanism of temperature regulation of solanesol accumulation.

The effects of in vitro batch digestion on water-in-oil-in-water (W/O/W) double emulsions encapsulated with anthocyanins (ACNs) from grape skin were investigated. The double emulsions exhibited the monomodal distribution (d = 686 ± 25 nm) showing relatively high encapsulation efficiency (87.74 ± 3.12%). After in vitro mouth digestion, the droplet size (d = 771 ± 26 nm) was significantly increased (p < 0.05). The double W1/O/W2 emulsions became a single W1/O emulsion due to proteolysis, which were coalesced together to form big particles with significant increases (p < 0.01) of average droplet sizes (d > 5 µm) after gastric digestion. During intestinal digestion, W1/O droplets were broken to give empty oil droplets and released ACNs in inner water phase, and the average droplet sizes (d < 260 nm) decreased significantly (p < 0.05). Our results indicated that ACNs were effectively protected by W/O/W double emulsions against in vitro mouth digestion and gastric, and were delivered in the simulated small intestine phase.

Blueberry pomace is a rich source of high-value bioactive polyphenols with presumed health benefits. Their incorporation into functional foods and health-related products benefits from coencapsulation and protection of polyphenol-rich extracts in suitable carriers. This study aimed to create a water-in-oil-in-water (W1/O/W2) double emulsion system suitable for the coencapsulation of total phenolics (TP) and anthocyanins (TA) from a polyphenol-rich extract of blueberry pomace (W1). The effect of critical physical parameters for preparing stable double emulsions, namely homogenization pressure, stirring speed and time, was investigated by measuring the hydrodynamic diameter, size dispersity and zeta potential of the oil droplets, and the encapsulation efficiency of TP and TA. The oil droplets were negatively charged (negative zeta potential values), which was related to the pH and composition of W2 (whey protein isolate solution) and suggests stabilization by the charged whey proteins. Increasing W1/O/W2 microfluidization pressure from 50 to 200 MPa or homogenization speed from 6000 to 12,000 rpm significantly increased droplet diameter and zeta potential and decreased TA and TP encapsulation efficiency. Increasing W1/O/W2 homogenization time from 15 to 20 min also increased droplet diameter and zeta potential and lowered TA encapsulation efficiency, while TP encapsulation did not vary significantly. In contrast, increasing W1/O homogenization time from 5 to 10 min at 10,000 rpm markedly increased TA encapsulation efficiency and reduced droplet diameter and zeta potential. High coencapsulation rates of blueberry polyphenols and anthocyanins around 80% or greater were achieved when the oil droplets were relatively small (mean diameter < 400 nm), with low dispersity (

The increase in diet-related chronic diseases has prompted the search for health-promoting compounds and methods to ensure their quality. Blueberry pomace is a rich yet underutilized source of bioactive polyphenols. For these high-value bioactive molecules, ultrasound-assisted extraction (USAE) is an attractive and green alternative to conventional extraction techniques for improving purity and yields. This study aimed to assess the impact of USAE parameters (sonication time, solvent composition, solid/liquid ratio, pH and temperature) on the recovery of phenolic compounds from blueberry pomace and antioxidant activity of the extracts. Total phenolic, flavonoid and anthocyanin contents (TPC, TFC and TAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity were analysed. USAE in 50% ethanol/water was the most efficient, yielding the highest TPC (22.33 mg/g dry matter (DM)), TFC (19.41 mg/g DM), TAC (31.32 mg/g DM) and DPPH radical scavenging activity (41.79 mg Trolox/g DM). USAE in water showed the lowest values even at low (1/40) solid/liquid ratio (7.85 mg/g DM, 3.49 mg/g DM, and 18.96 mg/g DM for TPC, TFC and TAC, respectively). Decreasing the solid/liquid ratio in water or 50% ethanol significantly increased TPC, TFC, TAC and DPPH radical scavenging. With ethanol, increasing the temperature in the range 20–40 °C decreased TPC but increased TFC and DPPH radical scavenging activity. Anthocyanin profiles of water and ethanolic extracts were qualitatively similar, consisting of malvidin, delphinidin, petunidin and cyanidin. These findings indicate that USAE is a method of choice for extracting high-value bioactive phenolics from blueberry pomace. Selective enrichment of different phenolic fractions is possible under select extraction conditions.

Zizania latifolia (tribe Oryzeae Dum., subfamily Oryzoideae Care, family Gramineae) is native to East Asian countries. The seeds of Z. latifolia (Chinese wild rice) have been consumed as a cereal in China for >3000 years. Z. latifolia forms swollen culms when infected with Ustilago esculenta, which is the second most-cultivated aquatic vegetable in China. The current review summarizes the nutrients and bioactive compounds of Z. latifolia, and health benefits of its seeds. The seeds of Z. latifolia contain proteins, minerals, vitamins, and bioactive compounds, the activities of which—for example, antioxidant activity—have been characterized. Various health benefits are associated with their consumption, such as alleviation of insulin resistance and lipotoxicity, and protection against cardiovascular disease. Chinese wild rice may be used to prevent and treat metabolic disease, such as diabetes, obesity, and cardiovascular diseases. Various compounds were isolated from the swollen culm, and aerial parts of Z. latifolia. The former suppresses osteoclast formation, inhibits growth of rat glioma cells, and may act as antioxidants and immunomodulators in drugs or foods. The latter exerts anti-fatigue, anti-inflammatory, and anti-allergic effects. Thus, Z. latifolia may be used to produce nutraceuticals and functional foods.

Sorghum grains with different applications had different phenolic profiles, which were corresponded to various antioxidant capacities. In this study, total phenolic, proanthocyanidins and flavonoids contents, as well as contents of individual phenolic compounds from sorghum grains with various applications were determined, and their antioxidant capacities were evaluated. Total phenolic contents (TPC) and total proanthocyanidins contents (TPAC) showed strong correlation with antioxidant activities (r > 0.95, p < 0.01). Hongyingzi (S-1), one of the brewing sorghums, showed the highest level of TPC and TPAC, while white grain sorghum (S-8) had the lowest. Except for black grain sorghum (S-7), that contained the highest contents of ferulic acid, brewing sorghum grains contained the higher contents of the most individual phenolic compounds, especially the variety S-1. The correlation among individual phenolic compounds and antioxidant activities indicated that the free forms of protocatechuic acid (r = 0.982 of FRAPassay, p < 0.01) and taxifolin (r = 0.826 of FRAP assay, p < 0.01) may be the main functional compounds. These results indicate that brewing sorghum grains can also be utilized as effective materials for functional foods.

Pectin had been recovered from canning wastewater produced by chemical treatment of segment membrane during preparation of canned citrus in our previous research. The purpose of this study was to characterize the extracted pectin from canning wastewater, and to evaluate its application as a fat alternative to replace fat in ice cream. The monosaccharide composition and rheological properties of the pectin were determined. The influences of fat reduction and pectin addition on the physicochemical, rheological and sensory properties of low-fat ice cream were determined. The rheological results showed that pectin solutions were typical pseudoplastic fluids. The addition of pectin in ice cream can cause an increase in viscosity, overrun, and hardness, and a decrease in meltdown of the ice cream. When 0.72% pectin (w/w) is incorporated into ice cream, a prototype product of ice cream with 45% lower fat content compared to the control was made. Results indicated that their qualities such as appearance, flavor, and taste were not significantly different. The low-fat ice cream had higher smoothness scores and lower mouth-coating scores. Hence, pectin extracted from citrus canning wastewater can be potentially used as fat replacer in ice cream, which benefits both the environment and the food industry.

Solanesol is a noncyclic terpene alcohol that is composed of nine isoprene units and mainly accumulates in solanaceous plants, especially tobacco (Nicotiana tabacum L.). In the present study, RNA-seq analyses of tobacco leaves, stems, and roots were used to identify putative solanesol biosynthesis genes. Six 1-deoxy-d-xylulose 5-phosphate synthase (DXS), two 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), two 2-C-methyl-d-erythritol 4-phosphate cytidylyltransferase (IspD), four 4-diphosphocytidyl-2-C-methyl-d-erythritol kinase (IspE), two 2-C-methyl-d-erythritol 2,4-cyclo-diphosphate synthase (IspF), four 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase (IspG), two 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (IspH), six isopentenyl diphosphate isomerase (IPI), and two solanesyl diphosphate synthase (SPS) candidate genes were identified in the solanesol biosynthetic pathway. Furthermore, the two N. tabacum SPS proteins (NtSPS1 and NtSPS2), which possessed two conserved aspartate-rich DDxxD domains, were highly homologous with SPS enzymes from other solanaceous plant species. In addition, the solanesol contents of three organs and of leaves from four growing stages of tobacco plants corresponded with the distribution of chlorophyll. Our findings provide a comprehensive evaluation of the correlation between the expression of different biosynthesis genes and the accumulation of solanesol, thus providing valuable insight into the regulation of solanesol biosynthesis in tobacco.

Abstract Autosomal Dominant Polycystic Kidney Disease (ADPKD) results in progressive cysts that lead to kidney failure, and is caused by heterozygous germline variants in PKD1 or PKD2. Cyst pathogenesis is not definitively understood. Somatic second-hit mutations have been implicated in cyst pathogenesis, though technical sequencing challenges have limited investigation. We used unique molecular identifiers, high-depth massively parallel sequencing and custom analysis techniques to identify somatic second-hit mutations in 24 whole cysts from disparate regions of six human ADPKD kidneys, utilising replicate samples and orthogonal confirmation. Average depth of coverage of 1166 error-corrected reads for PKD1 and 539 reads for PKD2 was obtained. 58% (14/24) of cysts had a detectable PKD1 somatic variant, with 5/6 participants having at least one cyst with a somatic variant. We demonstrate that low-frequency somatic mutations are detectable in a proportion of cysts from end-stage ADPKD human kidneys. Further studies are required to understand the drivers of this somatic mutation.