Cyanogenic Glucosides Research Articles

CRISPR-Cas9-mediated knockout of CYP79D1 and CYP79D2 in cassava attenuates toxic cyanogen production.

Cassava (Manihot esculenta) is a starchy root crop that supports over a billion people in tropical and subtropical regions of the world. This staple, however, produces the neurotoxin cyanide and requires processing for safe consumption. Excessive consumption of insufficiently processed cassava, in combination with protein-poor diets, can have neurodegenerative impacts. This problem is further exacerbated by drought conditions which increase this toxin in the plant. To reduce cyanide levels in cassava, we used CRISPR-mediated mutagenesis to disrupt the cytochrome P450 genes CYP79D1 and CYP79D2 whose protein products catalyze the first step in cyanogenic glucoside biosynthesis. Knockout of both genes eliminated cyanide in leaves and storage roots of cassava accession 60444; the West African, farmer-preferred cultivar TME 419; and the improved variety TMS 91/02324. Although knockout of CYP79D2 alone resulted in significant reduction of cyanide, mutagenesis of CYP79D1 did not, indicating these paralogs have diverged in their function. The congruence of results across accessions indicates that our approach could readily be extended to other preferred or improved cultivars. This work demonstrates cassava genome editing for enhanced food safety and reduced processing burden, against the backdrop of a changing climate.

The saponin bomb: a nucleolar-localized β-glucosidase hydrolyzes triterpene saponins in Medicago truncatula.

Plants often protect themselves from their own bioactive defense metabolites by storing them in less active forms. Consequently, plants also need systems allowing correct spatiotemporal reactivation of such metabolites, for instance under pathogen or herbivore attack. Via co-expression analysis with public transcriptomes, we determined that the model legume Medicago truncatula has evolved a two-component system composed of a β-glucosidase, denominated G1, and triterpene saponins, which are physically separated from each other in intact cells. G1 expression is root-specific, stress-inducible, and coregulated with that of the genes encoding the triterpene saponin biosynthetic enzymes. However, the G1 protein is stored in the nucleolus and is released and united with its typically vacuolar-stored substrates only upon tissue damage, partly mediated by the surfactant action of the saponins themselves. Subsequently, enzymatic removal of carbohydrate groups from the saponins creates a pool of metabolites with an increased broad-spectrum antimicrobial activity. The evolution of this defense system benefited from both the intrinsic condensation abilities of the enzyme and the bioactivity properties of its substrates. We dub this two-component system the saponin bomb, in analogy with the mustard oil and cyanide bombs, commonly used to describe the renowned β-glucosidase-dependent defense systems for glucosinolates and cyanogenic glucosides.

Species-specific dynamics of specialized metabolism in germinating sorghum grain revealed by temporal and tissue-resolved transcriptomics and metabolomics

Seed germination is crucial for plant productivity, and the biochemical changes during germination affect seedling survival, plant health and yield. While the general metabolism of germination is extensively studied, the role of specialized metabolism is less investigated. We therefore analyzed the metabolism of the defense compound dhurrin during sorghum (Sorghum bicolor) grain germination and early seedling development. Dhurrin is a cyanogenic glucoside, which is catabolized into different bioactive compounds at other stages of plant development, but its fate and role during germination is unknown. We dissected sorghum grain into three different tissues and investigated dhurrin biosynthesis and catabolism at the transcriptomic, metabolomic and biochemical level. We further analyzed transcriptional signature differences of cyanogenic glucoside metabolism between sorghum and barley (Hordeum vulgare), which produces similar specialized metabolites. We found that dhurrin is de novo biosynthesized and catabolized in the growing embryonic axis as well as the scutellum and aleurone layer, two tissues otherwise mainly acknowledged for their involvement in release and transport of general metabolites from the endosperm to the embryonic axis. In contrast, genes encoding cyanogenic glucoside biosynthesis in barley are exclusively expressed in the embryonic axis. Glutathione transferase enzymes (GSTs) are involved in dhurrin catabolism and the tissue-resolved analysis of GST expression identified new pathway candidate genes and conserved GSTs as potentially important in cereal germination. Our study demonstrates a highly dynamic tissue- and species-specific specialized metabolism during cereal grain germination, highlighting the importance of tissue-resolved analyses and identification of specific roles of specialized metabolites in fundamental plant processes.

The role of host plant usage and the accumulation of toxic secondary chemical compounds across the life cycle of a passion vine specialist butterfly

Abstract Host plant specialisation by herbivorous insects is ubiquitous, especially among the Lepidoptera. Many taxa have the ability to accumulate toxic compounds from their host plants that serve as chemical defences against natural enemies. Despite common knowledge of this pattern among insect ecologists, we still have much to learn about how dietary variation affects an insect's ability to acquire toxic plant chemicals. Longwing butterfly larvae (Heliconius) accumulate toxic cyanogenic glucosides from their host plants, passion vines (Passiflora), that make them toxic to most predators. Here, we present on zebra longwing (H. charithonia) caterpillar cohorts that were reared on native P. affinis, P. biflora, P. lutea, or P. suberosa to determine how host usage affected cyanide accumulation over larval developmental stages, and ultimately the toxicity of adult butterflies. Samples were collected at third, fourth and fifth instars, and teneral adults. Cyanogenic glucosides were extracted, the cyanide molecules were captured as sodium cyanide (CN), and CN content was quantified colorimetrically. The quantity of CN acquired increased linearly over larval ontogeny and was variable among host plants. CN concentration increased over larval development for two hosts but declined on the third. There was no clear relationship between host plant and adult CN concentrations, but adults that consumed certain hosts as larvae ended up accumulating significantly more CN, and thus expressed higher CN concentrations, than others. We demonstrated that some host plants provide specialist insects with more toxins than others and that larval secondary chemical accumulation may predict which life stages are more vulnerable to natural enemies. These conclusions support the hypothesis that host choice affects the survival of specialist insect herbivores.

Interactions between environmental factors drive selection on cyanogenesis in Trifolium repens

Phenotypic and genetic clines frequently evolve due to varying selection along environmental gradients. However, the specific environmental factors that impose differential selection are often multivariate and difficult to tease apart. We addressed this complexity using a factorial manipulation of watering, soil nutrients and (simulated) herbivory in controlled conditions to better understand the agents of selection driving the evolution of clines in a polymorphic chemical antiherbivore defence, hydrogen cyanide (HCN), of the plant white clover Trifolium repens. We found the presence or absence of the two metabolic components required for HCN production (cyanogenic glucosides and linamarase) to be more prominent determinants of vegetative growth and sexual reproduction in T. repens than HCN itself. We also found that the ability to produce one or both of cyanogenic glucosides or linamarase resulted in a growth advantage under drought and simulated herbivory that outweighs the metabolic cost of their production. These results support the view that the metabolic components underlying HCN play important roles beyond defence by increasing plant tolerance to stress. The growth advantage under drought, however, was diminished in the absence of nutrient addition, consistent with multivariate interactions as drivers of selection. This study provides novel insight into how a cosmopolitan plant has adapted to environmental gradients, and more generally, highlights the importance of considering interactions between multiple environmental factors when studying the evolution of phenotypic and genetic clines.

Probiotic and Cyanide Degrading Potentials of Pediococcus Pentosaceus and Pichia Exigua Isolated from Cassava Products Effluent

ABSTRACT Cassava tubers contain high levels of cyanogenic glucosides that are toxic when consumed. Lactic acid bacteria and yeast from the effluent generated during the processing of cassava products were assessed for cyanide degradation and probiotic potentials. Pediococcus sp. OG08 and Pichia sp. OG4 had higher bile salt tolerance (p < 0.05) in 0.3 % bile salts (82.5 %,77.7 %) and 0.5 % bile salts (69.3 %, 71.2 %) respectively. They displayed high potassium cyanide tolerance (Pediococcus sp. OG08, 45.7 % and Pichia sp. OG4, 44.7 %). Pediococcus pentosaceus OG08 and Pichia exigua OG4 resulted in the lowest residual cyanide content of 2.75 and 2.80 mg/kg respectively. The unfermented cassava flour had a residual cyanide content of 19.78 mg/kg. Pediococcus pentosaceus OG08 and Pichia exigua OG4 passed safety assessments, were valuable as probiotics due to their beneficial attributes. Cassava effluent possess valuable resource for potential probiotics.

Recruitment of distinct UDP-glycosyltransferase families demonstrates dynamic evolution of chemical defense within Eucalyptus L'Hér.

The economic and ecologically important genus Eucalyptus is rich in structurally diverse specialized metabolites. While some specialized metabolite classes are highly prevalent across the genus, the cyanogenic glucoside prunasin is only produced by c. 3% of species. To investigate the evolutionary mechanisms behind prunasin biosynthesis in Eucalyptus, we compared de novo assembled transcriptomes, together with online resources between cyanogenic and acyanogenic species. Identified genes were characterized invivo and invitro. Pathway characterization of cyanogenic Eucalyptus camphora and Eucalyptus yarraensis showed for the first time that the final glucosylation step from mandelonitrile to prunasin is catalyzed by a novel UDP-glucosyltransferase UGT87. This step is typically catalyzed by a member of the UGT85 family, including in Eucalyptus cladocalyx. The upstream conversion of phenylalanine to mandelonitrile is catalyzed by three cytochrome P450 (CYP) enzymes from the CYP79, CYP706, and CYP71 families, as previously shown. Analysis of acyanogenic Eucalyptus species revealed the loss of different ortholog prunasin biosynthetic genes. The recruitment of UGTs from different families for prunasin biosynthesis in Eucalyptus demonstrates important pathway heterogeneities and unprecedented dynamic pathway evolution of chemical defense within a single genus. Overall, this study provides relevant insights into the tremendous adaptability of these long-lived trees.

Development and Fecundity of Oriental Fruit Moth (Lepidoptera: Tortricidae) Reared on Various Concentrations of Amygdalin

Grapholita molesta (Busck) (Lepidoptera: Tortricidae), Oriental fruit moth (OFM), attacks fruits and shoots of the economically important trees in Rosaceae. Amygdalin is a cyanogenic glucoside of rosaceous plants that may be related to the seasonal patterns of infestation in many pests. The amygdalin concentration of fruits and shoots of peach, pear, and apple varies over the growing season. However, the relationship between the amygdalin concentration and G. molesta performance has not been reported. Here, we measured the performance (feeding, growth, development, and fecundity) of G. molesta larvae (as subsequent adults) reared on artificial diets with six amygdalin concentrations (0, 3, 6, 12, 24, and 48 mg/g), and we then calculated the population parameters. We found that these different concentrations of amygdalin affected the developmental time and fecundity, except for the proportion of larvae feeding on the diet and the survival rates of larvae and pupae. When compared with the control diet without amygdalin, diets with 3 or 6 mg/g (low and moderate concentrations) of amygdalin shortened developmental times and increased the number of eggs laid by females; however, a diet with 12 mg/g (moderate concentration) of amygdalin only increased the number of eggs laid by females and did not affect the larval and pupal developmental rate. A diet with 48 mg/g (high concentration) of amygdalin prolonged developmental times and reduced the number of eggs laid by females when compared with the control diet without amygdalin. Furthermore, the intrinsic rate of increase (rm) for insects reared on diets with 3 or 6 mg/g (low and moderate concentrations) of amygdalin versus the control diet without amygdalin showed a slightly improved population growth. However, this increase in the rm value did not persist over ten successive generations of rearing on the same diet. We concluded that the diet with 6 mg of amygdalin per g of diet can enhance the performance and population growth of G. molesta, but the effects of amygdalin are concentration-dependent.

Transcriptome analysis of the pulp of citrus fruitlets suggests that domestication enhanced growth processes and reduced chemical defenses increasing palatability

To identify key traits brought about by citrus domestication, we have analyzed the transcriptomes of the pulp of developing fruitlets of inedible wild Ichang papeda (Citrus ichangensis), acidic Sun Chu Sha Kat mandarin (C. reticulata) and three palatable segregants of a cross between commercial Clementine (C. x clementina) and W. Murcott (C. x reticulata) mandarins, two pummelo/mandarin admixtures of worldwide distribution. RNA-seq comparison between the wild citrus and the ancestral sour mandarin identified 7267 differentially expressed genes, out of which 2342 were mapped to 117 KEGG pathways. From the remaining genes, a set of 2832 genes was functionally annotated and grouped into 45 user-defined categories. The data suggest that domestication promoted fundamental growth processes to the detriment of the production of chemical defenses, namely, alkaloids, terpenoids, phenylpropanoids, flavonoids, glucosinolates and cyanogenic glucosides. In the papeda, the generation of energy to support a more active secondary metabolism appears to be dependent upon upregulation of glycolysis, fatty acid degradation, Calvin cycle, oxidative phosphorylation, and ATP-citrate lyase and GABA pathways. In the acidic mandarin, downregulation of cytosolic citrate degradation was concomitant with vacuolar citrate accumulation. These changes affected nitrogen and carbon allocation in both species leading to major differences in organoleptic properties since the reduction of unpleasant secondary metabolites increases palatability while acidity reduces acceptability. The comparison between the segregants and the acidic mandarin identified 357 transcripts characterized by the occurrence in the three segregants of additional downregulation of secondary metabolites and basic structural cell wall components. The segregants also showed upregulation of genes involved in the synthesis of methyl anthranilate and furaneol, key substances of pleasant fruity aroma and flavor, and of sugar transporters relevant for sugar accumulation. Transcriptome and qPCR analysis in developing and ripe fruit of a set of genes previously associated with citric acid accumulation, demonstrated that lower acidity is linked to downregulation of these regulatory genes in the segregants. The results suggest that the transition of inedible papeda to sour mandarin implicated drastic gene expression reprograming of pivotal pathways of the primary and secondary metabolism, while palatable mandarins evolved through progressive refining of palatability properties, especially acidity.

Whole-genome analysis guided molecular mechanism of cyanogenic glucoside degradation by yeast isolated from Prunus mume fruit syrup

Consumption of fermented Prunus mume fruit (maesil) sugar syrup raise a health concern due to the presence of the cyanogenic glucoside amygdalin. The goal of the present study was to investigate the amygdalin degradation potential and genome profile of the native microbes found in maesil syrup. The microbial profile analysis revealed that yeast is the predominant microorganism native to maesil syrup and that the isolated yeast cells showed a remarkable potential for amygdalin reduction (99.7%). Moreover, the reduction in amygdalin was inversely proportional to the growth of the isolated yeast. The whole-genome analysis revealed that the isolated yeast is Zygosaccharomyces rouxii (genome size 10 Mb, 39.25% of GC content). Of the 5250 genes (64.88%) predicted in the Z. rouxii genome, 5245 (99.90%) were annotated using NCBI Non-Redundant, UniProt, and InterProScan databases. The genome of the isolated Z. ruoxii harbored 2.03% of repeats and 0.68% of non-coding RNAs. Protein prediction indicated that β-glycosidases and hydroxynitrile lyase may play a key role in amygdalin degradation. The predicted degradation initiated by β-glycosidases that hydrolyze α-glucosidic bonds of amygdalin results in α-hydroxy nitriles (cyanohydrins) that are subsequently converted into carbonyl compounds (benzaldehyde) and hydrogen cyanide catalyzed by hydroxynitrile lyases. Present findings provide valuable data for constructing engineered microorganisms that can degrade amygdalin. Further analysis of Z. rouxii may elucidate the exact mechanism of amygdalin reduction in the production of maesil syrup.

Structure-guided engineering of key amino acids in UGT85B1 controlling substrate and stereo-specificity in aromatic cyanogenic glucoside biosynthesis.

SUMMARYCyanogenic glucosides are important defense molecules in plants with useful biological activities in animals. Their last biosynthetic step consists of a glycosylation reaction that confers stability and increases structural diversity and is catalyzed by the UDP‐dependent glycosyltransferases (UGTs) of glycosyltransferase family 1. These versatile enzymes have large and varied substrate scopes, and the structure–function relationships controlling scope and specificity remain poorly understood. Here, we report substrate‐bound crystal structures and rational engineering of substrate and stereo‐specificities of UGT85B1 from Sorghum bicolor involved in biosynthesis of the cyanogenic glucoside dhurrin. Substrate specificity was shifted from the natural substrate (S)‐p‐hydroxymandelonitrile to (S)‐mandelonitrile by combining a mutation to abolish hydrogen bonding to the p‐hydroxyl group with a mutation to provide steric hindrance at the p‐hydroxyl group binding site (V132A/Q225W). Further, stereo‐specificity was shifted from (S) to (R) by substituting four rationally chosen residues within 6 Å of the nitrile group (M312T/A313T/H408F/G409A). These activities were compared to two other UGTs involved in the biosynthesis of aromatic cyanogenic glucosides in Prunus dulcis (almond) and Eucalyptus cladocalyx. Together, these studies enabled us to pinpoint factors that drive substrate and stereo‐specificities in the cyanogenic glucoside biosynthetic UGTs. The structure‐guided engineering of the functional properties of UGT85B1 enhances our understanding of the evolution of UGTs involved in the biosynthesis of cyanogenic glucosides and will enable future engineering efforts towards new biotechnological applications.

Cocktails of Mycotoxins, Phytoestrogens, and Other Secondary Metabolites in Diets of Dairy Cows in Austria: Inferences from Diet Composition and Geo-Climatic Factors.

Dairy production is a pivotal economic sector of Austrian and European agriculture. Dietary toxins and endocrine disruptors of natural origin such as mycotoxins and phytoestrogens can affect animal health, reproduction, and productivity. This study characterized the profile of a wide spectrum of fungal, plant, and unspecific secondary metabolites, including regulated, emerging, and modified mycotoxins, phytoestrogens, and cyanogenic glucosides, in complete diets of lactating cows from 100 Austrian dairy farms. To achieve this, a validated multi-metabolite liquid chromatography/electrospray ionization–tandem mass spectrometric (LC/ESI–MS/MS) method was employed, detecting 155 of >800 tested metabolites. Additionally, the most influential dietary and geo-climatic factors related to the dietary mycotoxin contamination of Austrian dairy cattle were recognized. We evidenced that the diets of Austrian dairy cows presented ubiquitous contamination with mixtures of mycotoxins and phytoestrogens. Metabolites derived from Fusarium spp. presented the highest concentrations, were the most recurrent, and had the highest diversity among the detected fungal compounds. Zearalenone, deoxynivalenol, and fumonisin B1 were the most frequently occurring mycotoxins considered in the EU legislation, with detection frequencies >70%. Among the investigated dietary factors, inclusion of maize silage (MS) and straw in the diets was the most influential factor in contamination with Fusarium-derived and other fungal toxins and metabolites, and temperature was the most influential among the geo-climatic factors.

Regulation of cyanogenic glucosides in wild and domesticated Eusorghum taxa.

Domesticated sorghum (Sorghum bicolor [L.] Moench subsp. bicolor) diverts significant amounts of nitrogen away from primary metabolism to the synthesis of cyanogenic glucosides (CNglc) - specialized metabolites that release toxic hydrogen cyanide (HCN). Our aim was to identify the point in the genus Sorghum Moench at which plants gained the ability to maintain hazardous concentrations of cyanogenic glucosides in their leaves into maturity (HCN potential >0.4 mg g-1 ). This ability occurs in domesticated sorghum (in the subgenus Eusorghum), but not in wild taxa in other Sorghum subgenera. Eight accessions from the subgenus Eusorghum were grown in a common garden: an improved sorghum line, five sorghum landraces, the crop's wild progenitor (S. bicolor subsp. verticilliflorum [Steud.] de Wet ex Wiersema & J. Dahlb.) and wild Sorghum propinquum (Kunth) Hitchc. HCN potential was measured in plants (n = 80) at the three-leaf stage and at 6 weeks old. All study accessions, including the wild taxa, had hazardous CNglc concentrations in the leaves at both the three-leaf stage (mean HCN potentials > = 2.5 mg g-1 ) and at 6 weeks old (mean HCN potentials > = 0.68 mg g-1 ), greatly contrasting the much lower mature leaf HCN potentials previously found in wild Sorghum taxa outside subgenus Eusorghum (generally <= 0.01 mg g-1 ). Our results suggest that the ability to maintain hazardous leaf HCN potentials into maturity might have arisen during the divergence of Eusorghum from other Sorghum subgenera, rather than during the speciation or domestication of S. bicolor, and highlights the value of utilizing Sorghum taxa outside Eusorghum in efforts to improve the crop safety of sorghum.

Systematic Analysis of bHLH Transcription Factors in Cassava Uncovers Their Roles in Postharvest Physiological Deterioration and Cyanogenic Glycosides Biosynthesis.

The basic helix-loop-helix (bHLH) proteins are a large superfamily of transcription factors, and play a central role in a wide range of metabolic, physiological, and developmental processes in higher organisms. However, systematic investigation of bHLH gene family in cassava (Manihot esculenta Crantz) has not been reported. In the present study, we performed a genome-wide survey and identified 148 MebHLHs genes were unevenly harbored in 18 chromosomes. Through phylogenetic analyses along with Arabidopsis counterparts, these MebHLHs genes were divided into 19 groups, and each gene contains a similar structure and conserved motifs. Moreover, many cis-acting regulatory elements related to various defense and stress responses showed in MebHLH genes. Interestingly, transcriptome data analyses unveiled 117 MebHLH genes during postharvest physiological deterioration (PPD) process of cassava tuberous roots, while 65 MebHLH genes showed significantly change. Meanwhile, the relative quantitative analysis of 15 MebHLH genes demonstrated that they were sensitive to PPD, suggesting they may involve in PPD process regulation. Cyanogenic glucosides (CGs) biosynthesis during PPD process was increased, silencing of MebHLH72 and MebHLH114 showed that linamarin content was significantly decreased in the leaves. To summarize, the genome-wide identification and expression profiling of MebHLH candidates pave a new avenue for uderstanding their function in PPD and CGs biosynthesis, which will accelerate the improvement of PPD tolerance and decrease CGs content in cassava tuberous roots.

Condition dependence in biosynthesized chemical defenses of an aposematic and mimetic Heliconius butterfly.

Aposematic animals advertise their toxicity or unpalatability with bright warning coloration. However, acquiring and maintaining chemical defenses can be energetically costly, and consequent associations with other important traits could shape chemical defense evolution. Here, we have tested whether chemical defenses are involved in energetic trade‐offs with other traits, or whether the levels of chemical defenses are condition dependent, by studying associations between biosynthesized cyanogenic toxicity and a suite of key life‐history and fitness traits in a Heliconius butterfly under a controlled laboratory setting. Heliconius butterflies are well known for the diversity of their warning color patterns and widespread mimicry and can both sequester the cyanogenic glucosides of their Passiflora host plants and biosynthesize these toxins de novo. We find energetically costly life‐history traits to be either unassociated or to show a general positive association with biosynthesized cyanogenic toxicity. More toxic individuals developed faster and had higher mass as adults and a tendency for increased lifespan and fecundity. These results thus indicate that toxicity level of adult butterflies may be dependent on individual condition, influenced by genetic background or earlier conditions, with maternal effects as one strong candidate mechanism. Additionally, toxicity was higher in older individuals, consistent with previous studies indicating accumulation of toxins with age. As toxicity level at death was independent of lifespan, cyanogenic glucoside compounds may have been recycled to release resources relevant for longevity in these long‐living butterflies. Understanding the origins and maintenance of variation in defenses is necessary in building a more complete picture of factors shaping the evolution of aposematic and mimetic systems.

Cyanide Content of Cassava Food Products Available in Australia

In 2009, Food Standards Australia New Zealand set a total cyanide content limit of 10 ppm for ready-to-eat cassava products to address food safety concerns about cyanogenic glucosides in cassava. This study surveys a range of cassava food products available in Melbourne, Australia, ten years after the implementation of these regulations. Of all the products tested, the mean cyanide content was greatest in ready-to-eat cassava chips (48.4 ppm), although imported ready-to-eat products had a higher mean cyanide content (95.9 ppm) than those manufactured in Australia (1.0 ppm). Cyanide was detected in frozen cassava products (grated mean = 12.9 ppm; whole root mean = 19.8 ppm), but was significantly reduced through processing according to packet instructions in both product types. Three methods were used to quantify total cyanide content: the evolved cyanide method, the picrate absorbance method and the picrate chart method, with satisfactory agreement between methods. The picrate absorbance and chart methods reported mean cyanide contents 13.7 ppm and 23.1 ppm higher, respectively, than the evolved cyanide method. Our results reaffirm the need for the ongoing testing of cassava food products, especially ready-to-eat products whose cyanide content will not be reduced before consumption.

What’s your poison? Cyanide production regulated by a bHLH transcription factor inLotus japonicus

Humans hold a love–hate relationship with cyanogenic plants. Preferentially selected for domestication due to their heightened pest resistance, they can nevertheless threaten both human and livestock health if not consumed in moderation or adequately processed. Over 20% of crops are cyanogenic due to the stable accumulation of cyanogenic glucosides (CNglcs) and, while the biochemical pathways of these specialized metabolites are known, the same cannot be said regarding their underlying molecular pathways. In a comprehensive study, Chen et al. (2022) characterized a basic helix–loop–helix (bHLH) transcription factor that is responsive to methyl jasmonate and directly induces expression of the key CNglc biosynthetic gene in the model species Lotus japonicus.

Metabolism of Aldoximes and Nitriles in Plant-Associated Bacteria and Its Potential in Plant-Bacteria Interactions.

In plants, aldoximes per se act as defense compounds and are precursors of complex defense compounds such as cyanogenic glucosides and glucosinolates. Bacteria rarely produce aldoximes, but some are able to transform them by aldoxime dehydratase (Oxd), followed by nitrilase (NLase) or nitrile hydratase (NHase) catalyzed transformations. Oxds are often encoded together with NLases or NHases in a single operon, forming the aldoxime–nitrile pathway. Previous reviews have largely focused on the use of Oxds and NLases or NHases in organic synthesis. In contrast, the focus of this review is on the contribution of these enzymes to plant-bacteria interactions. Therefore, we summarize the substrate specificities of the enzymes for plant compounds. We also analyze the taxonomic and ecological distribution of the enzymes. In addition, we discuss their importance in selected plant symbionts. The data show that Oxds, NLases, and NHases are abundant in Actinobacteria and Proteobacteria. The enzymes seem to be important for breaking through plant defenses and utilizing oximes or nitriles as nutrients. They may also contribute, e.g., to the synthesis of the phytohormone indole-3-acetic acid. We conclude that the bacterial and plant metabolism of aldoximes and nitriles may interfere in several ways. However, further in vitro and in vivo studies are needed to better understand this underexplored aspect of plant-bacteria interactions.

Dhurrin increases but does not mitigate oxidative stress in droughted Sorghum bicolor

Main conclusionDroughted sorghum had higher concentrations of ROS in both wildtype and dhurrin-lacking mutants. Dhurrin increased in wildtype genotypes with drought. Dhurrin does not appear to mitigate oxidative stress in sorghum.Sorghum bicolor is tolerant of high temperatures and prolonged droughts. During droughts, concentrations of dhurrin, a cyanogenic glucoside, increase posing a risk to livestock of hydrogen cyanide poisoning. Dhurrin can also be recycled without the release of hydrogen cyanide presenting the possibility that it may have functions other than defence. It has been hypothesised that dhurrin may be able to mitigate oxidative stress by scavenging reactive oxygen species (ROS) during biosynthesis and recycling. To test this, we compared the growth and chemical composition of S. bicolor in total cyanide deficient sorghum mutants (tcd1) with wild-type plants that were either well-watered or left unwatered for 2 weeks. Plants from the adult cyanide deficient class of mutant (acdc1) were also included. Foliar dhurrin increased in response to drought in all lines except tcd1 and acdc1, but not in the roots or leaf sheaths. Foliar ROS concentration increased in drought-stressed plants in all genotypes. Phenolic concentrations were also measured but no differences were detected. The total amounts of dhurrin, ROS and phenolics on a whole plant basis were lower in droughted plants due to their smaller biomass, but there were no significant genotypic differences. Up until treatments began at the 3-leaf stage, tcd1 mutants grew more slowly than the other genotypes but after that they had higher relative growth rates, even when droughted. The findings presented here do not support the hypothesis that the increase in dhurrin commonly seen in drought-stressed sorghum plays a role in reducing oxidative stress by scavenging ROS.

The Investigation of Phenylalanine, Glucosinolate, Benzylisothiocyanate (BITC) and Cyanogenic Glucoside of Papaya Fruits (Carica papaya L. cv. ‘Tainung No. 2’) under Different Development Stages between Seasons and Their Correlation with Bitter Taste

Papaya fruit is one of economic crops in Taiwan, mostly eaten as table fruits. In some Asian countries, unripe papaya fruit is eaten as salad and this led to trends in Taiwan as well. However, unripe papaya fruit may taste bitter during cool seasons. Glucosinolate and cyanogenic glucoside are among the substances that cause bitter taste in many plants, which can also be found in papaya. However, there is still no report about the relationship between seasons and bitter taste in papaya fruits. Thus, the purpose of this study is to investigate the glucosinolate biosynthesis and its correlation between bitterness intensity during cool and warm seasons. The bitterness intensity was highest at the young fruit stage and decreased as it developed. In addition, the bitterness intensity in cool season fruits is higher than in warm season fruits. Cyanogenic glucoside and BITC content showed negative correlation with bitterness intensity (r = −0.54 ***; −0.46 ***). Phenylalanine showed positive correlation with bitterness intensity (r = 0.35 ***), but its content did not reach the bitterness threshold concentration, which suggested that phenylalanine only acts as cyanogenic glucoside and glucosinolate precusors. Glucosinolate content showed positive correlation with bitterness intensity at different developmental stages (r = 0.805 ***). However, the correlation value in different lines/cultivars decreased (0.44 ***), suggesting that glucosinolate was not the only substance that caused bitter taste in immature papaya fruits.