Search Results (14)

Microgreens are increasingly promoted as sustainable, nutrient-dense foods, yet their content of potentially harmful specialized metabolites remains poorly explored. Here, we developed and applied a reversed-phase liquid chromatography–electrospray ionization high-resolution mass spectrometry (RPLC-ESI-HRMS) method for the detection of cyanogenic glycosides (CNGs) in edible microgreens. Method optimization, performed using dhurrin and lotaustralin as model standards, showed that positive ion detection of sodium adducts provided the most informative and selective HRMS/MS response, with diagnostic fragmentation patterns suitable for CNG recognition in complex matrices. Quantitative validation for lotaustralin showed excellent linearity (R² = 0.998), low detection/quantification limits (LOD 0.16 mg/L; LOQ 0.53 mg/L), good extraction recovery, and a negligible matrix effect. Application of the method revealed a clear species-dependent profile. No detectable CNGs were found in broccoli raab and kale microgreens, supporting their safety as ready-to-eat products in this respect. In contrast, flax microgreens contained four CNGs: linamarin, lotaustralin, linustatin, and neolinustatin. Monoglycosylated species predominated, with lotaustralin quantified at 5.5 ± 0.6 mg/g dry weight and linamarin estimated at even higher levels. Diglycosylated CNGs were present at much lower concentrations and displayed multiple chromatographic peaks, consistent with the occurrence of structurally related isomeric forms. These quantitative results are specific to the flax microgreen samples analyzed here, obtained by pooling the lyophilized material obtained from several plants; thus, they do not account for biological variability among individual plants. Based on the measured CNG levels, flax microgreens showed a non-negligible cyanogenic potential. Assuming 1, 10 and 25% conversion to hydrogen cyanide, the estimated release would be, respectively, about 3, 33 and 81 mg HCN/kg of fresh flax microgreens, values lower than the current EU limit (150 mg HCN/kg of edible product) for flaxseed intended for direct consumption but comparable to values reported for other foods. These findings highlight the need to complement the nutritional evaluation of novel microgreens with targeted toxicological screening. Full article

The sorghum aphid (Melanaphis sacchari (Zehntner, 1897)), a globally destructive pest, severely compromises sorghum yield and quality. This study compared aphid-resistant (HX133) and aphid-susceptible (HX37) sorghum (Sorghum bicolor (L.) Moench) cultivars, revealing that HX133 significantly suppressed aphid proliferation through repellent and antibiotic effects, while aphid populations increased continuously in HX37. Transcriptome analysis identified 2802 differentially expressed genes (DEGs, 45.9% upregulated) in HX133 at 24 h post-infestation, in contrast with only 732 DEGs (21% upregulated) in HX37. Pathway enrichment highlighted shikimate-mediated phenylpropanoid/flavonoid biosynthesis and glutathione metabolism as central to HX133’s defense response, alongside photosynthesis-related pathways common to both cultivars. qRT-PCR validation confirmed activation of the shikimate pathway in HX133, driving the synthesis of dhurrin—a cyanogenic glycoside critical for aphid resistance—and other tyrosine-derived metabolites (e.g., benzyl isoquinoline alkaloids, tocopherol). These findings demonstrate that HX133 employs multi-layered metabolic regulation, particularly dhurrin accumulation, to counteract aphid infestation, whereas susceptible cultivars exhibit limited defense induction. This work provides molecular targets for enhancing aphid resistance in sorghum breeding programs. Full article

The cyanogenic glucoside dhurrin is found in sorghum and has been reported for its role in defense against biotic and abiotic stresses, both involving hydrogen cyanide (HCN) release. The fungus Sporisorium reilianum f. sp. reilianum (SRS) causes sorghum head smut and the infection occurs at the seedling stage, later resulting in panicle loss. Here, the focus was to determine the role of dhurrin in sorghum’s reaction against SRS infection. We investigated the genomic basis of HCN potential (HCNp) variation and its relationship with seedlings’ response to SRS inoculation, along with other sorghum traits, and the expression of dhurrin biosynthetic genes in SRS-inoculated young sorghum. Genome-wide association studies (GWAS) using HCNp scores showed significant single nucleotide polymorphisms (SNPs) on chromosomes harboring the dhurrin biosynthetic and catabolic genes but not in proximity. Significant hits were also detected in or near genes encoding proteins involved in plant defense/resistance against biotic stresses. Correlation analyses showed a strong positive relationship between average HCNp scores and latent period in SRS-inoculated sorghum seedlings. RT-qPCR revealed that the dhurrin biosynthetic genes were upregulated in the leaves of the head smut resistant line BTx635 up to two days after SRS inoculation. Our results suggest the involvement of dhurrin in sorghum’s protection against SRS. Full article

Sorghum (Sorghum bicolor) holds a significant position as the fifth most vital cereal crop globally. Its drought resistance and robust biomass production, coupled with commendable nutritional value, make sorghum a promising choice for animal feed. Nevertheless, the utilization of sorghum in animal production faces hurdles of dhurrin (a cyanogenic glycoside) poisoning. While dhurrin serves as a protective secondary metabolite during sorghum growth, the resulting highly toxic hydrogen cyanide poses a significant threat to animal safety. This review extensively examines the biometabolic processes of dhurrin, the pivotal genes involved in the regulation of dhurrin biosynthesis, and the factors influencing dhurrin content in sorghum. It delves into the impact of dhurrin on animal production and explores measures to mitigate its content, aiming to provide insights for advancing research on dhurrin metabolism regulation in sorghum and its rational utilization in animal production. Full article

LC-MS/MS analyses have been reported as challenging for the reliable separation and quantification of cyanogenic glycosides (CNGs), especially (R)-prunasin and sambunigrin isomers found in American elderberry (Sambucus nigra L. subsp. canadensis (L.) Bolli). Hence, a novel multiple reaction monitoring (MRM)-based ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated in the present study for simultaneous separation and quantification of five CNGs, including amygdalin, dhurrin, linamarin, (R)-prunasin, and (S)-prunasin (commonly referred to as sambunigrin). Initially, the role of ammonium formate was investigated as an aqueous mobile-phase additive in developing MRM-based UHPLC-MS/MS. Later, chromatographic conditions for the resolved separation of (R)-prunasin and sambunigrin were identified. Validation studies confirmed that the developed method has good linearity and acceptable precision and accuracy. A noticeable matrix effect (mainly signal enhancement) was observed in leaf samples only. This method was used to detect and quantify CNGs, including (R)-prunasin and sambunigrin, in leaf and fruit samples of American elderberry. Among the studied CNGs, only (R)-prunasin was detected in the leaf samples. Interestingly, (S)-prunasin (sambunigrin) was not detected in the samples analyzed, even though it has been previously reported in elderberry species. Full article

Cyanogenic glucosides are specialized metabolites produced by over 3000 species of higher plants from more than 130 families. The deployment of cyanogenic glucosides is influenced by biotic and abiotic factors in addition to being developmentally regulated, consistent with their roles in plant defense and stress mitigation. Despite their ubiquity, very little is known regarding the molecular mechanisms that regulate their biosynthesis. The biosynthetic pathway of dhurrin, the cyanogenic glucoside found in the important cereal crop sorghum (Sorghum bicolor (L.) Moench), was described over 20 years ago, and yet no direct regulator of the biosynthetic genes has been identified. To isolate regulatory proteins that bind to the promoter region of the key dhurrin biosynthetic gene of sorghum, SbCYP79A1, yeast one-hybrid screens were performed. A bait fragment containing 1204 base pairs of the SbCYP79A1 5′ regulatory region was cloned upstream of a reporter gene and introduced into Saccharomyces cerevisiae. Subsequently, the yeast was transformed with library cDNA representing RNA from two different sorghum developmental stages. From these screens, we identified SbGATA22, an LLM domain B-GATA transcription factor that binds to the putative GATA transcription factor binding motifs in the SbCYP79A1 promoter region. Transient assays in Nicotiana benthamiana show that SbGATA22 localizes to the nucleus. The expression of SbGATA22, in comparison with SbCYP79A1 expression and dhurrin concentration, was analyzed over 14 days of sorghum development and in response to nitrogen application, as these conditions are known to affect dhurrin levels. Collectively, these findings suggest that SbGATA22 may act as a negative regulator of SbCYP79A1 expression and provide a preliminary insight into the molecular regulation of dhurrin biosynthesis in sorghum. Full article

Sorghum plants naturally produce dhurrin, a cyanogenic glycoside that may be hydrolysed to cyanide, resulting in often-lethal toxicoses. Ruminants are particularly sensitive to cyanogenic glycosides due to the active role of rumen microbiota in dhurrin hydrolysis. This work provides an overview of a poisoning outbreak that occurred in 5 farms in Northwest Italy in August 2022; a total of 66 cows died, and many others developed acute toxicosis after being fed on either cultivated (Sorghum bicolor) or wild Sorghum (Sorghum halepense). Clinical signs were recorded, and all cows received antidotal/supportive therapy. Dead animals were subjected to necropsy, and dhurrin content was determined in Sorghum specimens using an LC–MS/MS method. Rapid onset, severe respiratory distress, recumbency and convulsions were the main clinical features; bright red blood, a bitter almond smell and lung emphysema were consistently observed on necropsy. The combined i.v. and oral administration of sodium thiosulphate resulted in a rapid improvement of clinical signs. Dhurrin concentrations corresponding to cyanide levels higher than the tolerated threshold of 200 mg/kg were detected in sorghum specimens from 4 out of 5 involved farms; thereafter, such levels declined, reaching tolerable concentrations in September–October. Feeding cattle with wild or cultivated Sorghum as green fodder is a common practice in Northern Italy, especially in summer. However, care should be taken in case of adverse climatic conditions, such as severe drought and tropical temperatures (characterising summer 2022), which are reported to increase dhurrin synthesis and storage. Full article

Post-flowering drought tolerance (stay-green) in grain sorghum (Sorghum bicolor (L.) Moench) is an important agronomic trait in many arid and semiarid environments throughout the world. Stay-green has been associated with increased grain yields, as well as resistance to lodging and charcoal rot disease. Nonetheless, the relative effects of genotype, environment, and genotype × environment interactions are not well understood for this trait; similarly, the relationship between various leaf sugars and stay-green has not been sufficiently evaluated in diverse germplasm. Thus, the goals of this study were to determine the genotype, environment, and genotype by environment (GxE) effects for leaf dhurrin, sugars, and stay-green in ten diverse grain sorghum breeding lines, to evaluate the Pearson’s correlation coefficients (r) between these traits, and to determine entry-mean repeatability (R) for each of these traits. Of the compositional traits studied, we determined that leaf dhurrin had the highest correlation with the stay-green phenotypes (r = −0.62). We found that stay-green sorghum lines contained approximately 2–3 times as much dhurrin as non-stay-green lines, with B1778 containing the highest concentration of dhurrin (84.8 µg/cm²) and Tx7000 containing the least (20.9 µg/cm²). The differences between the environments for several of the traits were high, and all the traits examined had high repeatability (R = 0.89–0.92). These data demonstrate a relationship between leaf dhurrin and the stay-green phenotypes in sorghum, and further study will allow researchers to determine the causal effect that dhurrin has on post-flowering drought tolerance in sorghum. Full article

Sorghum bicolor L. is a vital cereal crop for global food security. Its adaptability to diverse climates make it economically, socially, and environmentally valuable. However, soil salinization caused by climate extremes poses a threat to sorghum. This study aimed to identify candidate salt-tolerant genes and single nucleotide polymorphisms (SNPs) by performing a comparative transcriptome analysis on a mutant sorghum line and its wild type. The mutant line was generated through gamma ray exposure and selection for salt tolerance. Phenotypic measurements were taken, followed by mRNA sequencing and variant calling. In this study, potential genes and non-synonymous SNPs associated with salt tolerance were inferred, including LOC8071970, LOC8067721, LOC110430887, LOC8070256, and LOC8056880. These genes demonstrated notable differences in nsSNPs in comparison to the wild type, suggesting their potential roles in salt tolerance. Additionally, LOC8060874 (cyanohydrin beta-glucosyltransferase) was suggested as a key gene involved in salt tolerance due to its possible role in dhurrin biosynthesis under salt stress. In upcoming research, additional reverse genetics studies will be necessary in order to verify the function of those candidate genes in relation to salt stress. In conclusion, this study underscores the significance of investigating salt tolerance mechanisms and the potential key genes associated with salt tolerance in sorghum. Our findings may provide insights for future breeding strategies aimed at enhancing salinity tolerance and crop productivity. Full article

Domestication has resulted in a loss of genetic diversity in our major food crops, leading to susceptibility to biotic and abiotic stresses linked with climate change. Crop wild relatives (CWR) may provide a source of novel genes potentially important for re-gaining climate resilience. Sorghum bicolor is an important cereal crop with wild relatives that are endemic to Australia. Sorghum bicolor is cyanogenic, but the cyanogenic status of wild Sorghum species is not well known. In this study, leaves of wild species endemic in Australia are screened for the presence of the cyanogenic glucoside dhurrin. The direct measurement of dhurrin content and the potential for dhurrin-derived HCN release (HCNp) showed that all the tested Australian wild species were essentially phenotypically acyanogenic. The unexpected low dhurrin content may reflect the variable and generally nutrient-poor environments in which they are growing in nature. Genome sequencing of six CWR and PCR amplification of the CYP79A1 gene from additional species showed that a high conservation of key amino acids is required for correct protein function and dhurrin synthesis, pointing to the transcriptional regulation of the cyanogenic phenotype in wild sorghum as previously shown in elite sorghum. Full article

Allelopathy is an ecological phenomenon that involves the production and release of biomolecules from different crops, cultivated plants, and bacteria or fungi into the soil rhizosphere and impacts other organisms in the vicinity. Sorghum possesses vital allelopathic characteristics due to which it produces and releases different biomolecules from its root hairs, stems, and grains. Several studies have reported that sorghum acts as an allelopathic crop, decreasing the growth and eco-physiological attributes of surrounding plants and weeds growing simultaneously or subsequently in the field. Sorghum allelopathy has been exploited in the context of green manure, crop rotations, cover crops, and intercropping or mulching, whereas plant aqueous extracts or powder might be an alternate method of weed control. A diverse group of allelochemicals, including benzoic acid, p-hydroxybenzoic acid, vanillic acid, ferulic acid, chlorogenic acid, m-coumaric acid, p-coumaric acid, gallic acid, caffeic acid, p-hydroxibenzaldehyde, dhurrin, sorgoleone, m-hydroxybenzoic acid and protocatechuic acid, have been isolated and identified from different plant tissues of sorghum and root exudates. These allelochemicals, especially sorgoleone, have been investigated in terms of their mode(s) of action, specific activity and selectivity, release in the rhizosphere and uptake and translocation in sensitive species. The present review describes the importance of sorghum allelopathy as an ecological tool in managing weeds, highlighting the most recent advances in the allelochemicals present in sorghum, their modes of action, and their fate in the ecosystem. Further research should focus on the evaluation and selection of sorghum cultivars with high allelopathic potential, so that sorghum allelopathy can be better utilized for weed control and yield enhancement. Full article

In plants, the production of secondary metabolites is considered to be at the expense of primary growth. Sorghum produces a cyanogenic glycoside (dhurrin) that is believed to act as its chemical defence. Studies have shown that acyanogenic plants are smaller in size compared to the wildtype. This study aimed to investigate whether the small plant size is due to delayed germination or due to the lack of dhurrin derived nitrogen. A novel plant system consisting of totally cyanide deficient class 1 (tcd1) and adult cyanide deficient 1 (acdc1) mutant lines was employed. The data for germination, plant height and developmental stage during seedling development and final plant reproductive fitness was recorded. The possible role of phytohormones in recovering the wildtype phenotype, especially in developmentally acyanogenic acdc1 line, was also investigated. The data on plant growth have shown that the lack of dhurrin is disadvantageous to growth, but only at the early developmental stage. The tcd1 plants also took longer to mature probably due to delayed flowering. None of the tested hormones were able to recover the wildtype phenotype. We conclude that the generation of dhurrin is advantageous for plant growth, especially at critical growth stages like germinating seed by providing a ready source of reduced nitrogen. Full article

Sorghum (Sorghum bicolor (L.)) Moench is an important food for humans and feed for livestock. Sorghum contains dhurrin which can be degraded into toxic hydrogen cyanide. Here, we report the expression patterns of 14 candidate genes related to dhurrin ((S)-4-Hydroxymandelnitrile-β-D-glucopyranoside) metabolism and the effects of the gene expression on specific metabolite content in selected sorghum accessions. Dhurrin-related metabolism is vigorous in the early stages of development of sorghum. The dhurrin contents of most accessions tested were in the range of approximately 6–22 μg mg⁻¹ fresh leaf tissue throughout growth. The p-hydroxybenzaldehyde (pHB) contents were high at seedling stages, but almost nonexistent at adult stages. The contents of p-hydroxyphenylacetic acid (pHPAAc) were relatively low throughout growth compared to those of dhurrin or pHB. Generally, the expression of the candidate genes was higher at seedling stage than at other stages and decreased gradually as plants grew. In addition, we identified significant SNPs, and six of them were potentially associated with non-synonymous changes in CAS1. Our results may provide the basis for choosing breeding materials to regulate cyanide contents in sorghum varieties to prevent HCN toxicity of livestock or to promote drought tolerance or pathogen resistance. Full article

Macadamia has increasing commercial importance in the food, cosmetics, and pharmaceutical industries. However, the toxic compound hydrogen cyanide (HCN) released from the hydrolysis of cyanogenic compounds in Macadamia causes a safety risk. In this study, optimum conditions for the maximum release of HCN from Macadamia were evaluated. Direct headspace analysis of HCN above Macadamia plant parts (flower, leaves, nuts, and husks) was carried out using selected ion flow tube–mass spectrometry (SIFT-MS). The cyanogenic glycoside dhurrin and total cyanide in the extracts were analyzed using HPLC-MS and UV–vis spectrophotometer, respectively. HCN released in the headspace was at a maximum when Macadamia samples were treated with pH 7 buffer solution and heated at 50 °C for 60 min. Correspondingly, treatment of Macadamia samples under these conditions resulted in 93–100% removal of dhurrin and 81–91% removal of total cyanide in the sample extracts. Hydrolysis of cyanogenic glucosides followed a first-order reaction with respect to HCN production where cyanogenesis is principally induced by pH changes initiating enzymatic hydrolysis rather than thermally induced reactions. The effective processing of different Macadamia plant parts is important and beneficial for the safe production and utilization of Macadamia-based products. Full article