Regulation of 2-Acetyl-1-pyrroline Content in Fragrant Rice under Different Temperatures at the Grain-Filling Stage.

2-Acetyl-1-pyrroline (2-AP) is a key volatile organic compound in fragrant rice aroma. However, the effects of temperature on 2-AP biosynthesis in fragrant rice and its regulation mechanism have been rarely reported. In the present study, three fragrant rice varieties were used as plant materials, and four temperature treatments during the grain-filling stage, i.e., (T1) 22/17 °C, (T2) 27/22 °C, (T3) 32/27 °C, and (T4) 37/32 °C, were adopted. The results showed that grain contents of 2-AP, proline, and γ-aminobutyric acid (GABA) significantly (P < 0.05) increased with decreased temperature, while the lowest and highest 2-AP contents were recorded in the T4 and T1 treatments, respectively. Higher pyrroline-5-carboxylic acid (P5C) content was recorded in low-temperature treatments (T1 and T2) than in high-temperature treatments (T3 and T4). The transcript levels of genes BADH2, PRODH, and OAT significantly (P < 0.05) decreased with decreased temperature. Lower transcript levels of genes P5CR, P5CS2, DAO2, DAO4, and DAO5 were recorded in low-temperature treatments (T1 and T2) than in high-temperature treatments (T3 and T4). In conclusion, low temperature increased 2-AP content and high temperature decreased 2-AP content in fragrant rice. We deduced that temperature regulated 2-AP biosynthesis through the metabolism of proline and GABA.

Development and validation of a QuEChERS-HPLC-DAD method using polymer-functionalized melamine sponges for the analysis of antipsychotic drugs in milk.

The prohibition of antipsychotic drugs in animal foodstuffs has raised significant concerns. In this study, a novel matrix purification adsorbent comprising a polymer (polyaniline and polypyrrole)-functionalized melamine sponge (Ms) was employed for the high performance liquid chromatography-diode array detector (HPLC-DAD) detection of three phenothiazines (chlorpromazine, thioridazine, and promethazine), and a tricyclic imipramine in milk. The as-prepared functionalized Ms was characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, and water contact angle measurements. Excellent linearity with a coefficient of determination (R2) of 0.999 was achieved for all drugs within the concentration range of 0.01-47.00 µg mL-1. The recoveries of the four analytes ranged from 92.1 % to 106.9 % at the three spiked levels. These results demonstrate the successful application of the proposed method for the determination of the four drugs. Cost-effective polymer-functionalized Ms is a viable alternative for matrix purification, enabling rapid determination of drug residues in diverse food samples.

Removal of an agricultural herbicide (2,4-Dichlorophenoxyacetic acid) using magnetic nanocomposite: A combined experimental and modeling studies.

This study focused on modeling the removal of one of the widely used agricultural herbicides known as 2,4-Dichlorophenoxyacetic acid (2,4-D) using polypyrrole-coated Fe2O3 nanoparticles (Fe2O3@PPy). The Fe2O3@PPy nanocomposite was synthesized by surface-coating the Tabebuia aurea leaf extract synthesized Fe2O3 nanoparticles with polypyrrole. After characterization, the adsorptive potential of the nanocomposite for removing 2,4-D from aqueous solution was examined. Central composite design (CCD) was employed for optimizing the adsorption, revealing an adsorption efficiency of 90.65% at a 2,4-D concentration of 12 ppm, a dosage of 3.8 g/L, an agitation speed of 150 rpm, and 196 min. Adsorption dataset fitted satisfactorily to Langmuir isotherm (R2: 0.984 & χ2: 0.054) and pseudo-second-order kinetics (R2: 0.929 & χ2: 0.013) whereas the exothermic and spontaneous nature were confirmed via the thermodynamic study. The predictive models, including adaptive neuro-fuzzy inference system (ANFIS), artificial neural network (ANN), and response surface methodology (RSM), demonstrated good precision for the prediction of 2,4-D adsorption, with respective R2 of 0.9719, 0.9604, and 0.9528. Nevertheless, statistical analysis supported ANFIS as the better forecasting tool, while RSM was the least effective. The maximum adsorption capacity of 2,4-D onto the Fe2O3@PPy nanocomposite was 7.29 mg/g, significantly higher than a few reported values. Therefore, the Fe2O3@PPy nanocomposite could serve as a competent adsorbent to remove 2,4-D herbicide from aqueous streams.

One-step synthesis of nitrogen-functionalized graphene aerogel for efficient removal of hexavalent chromium in water.

The efficient removal of hexavalent chromium (Cr(VI)) with high toxicity has attracted widespread concern since it causes serious harm to ecological environment and public health. Herein, we report a novel nitrogen-functionalized graphene aerogel with stereoscopic structure through a simple hydrothermal method and freeze drying for Cr(VI) removal from water. Graphene oxide (GO) and nitrogenous organics polyethyleneimine (PEI) and pyrrole are used as raw materials for preparing PEI/polypyrrole/GO aerogel (PPGA) adsorbent. PEI and pyrrole act as nitrogen sources to introduce nitrogenous functional groups, and also take on the role of cross-linkers for helping GO sheets to form stereoscopic structure. The obtained PPGA has a fast adsorption rate, excellent reusability, and shows a remarkable adsorption capacity for Cr(VI) up to 458.24 mg/g under the optimal conditions (pH 2.0, 298 K, Cr(VI): 600 mg/L, dosage: 0.4 g/L). The adsorption process of Cr(VI) on PPGA can fit the pseudo-second-order kinetic model well. Analysis of intraparticle diffusion shows that the Cr(VI) removal process is a multi-step process. Adsorption thermodynamic and isotherm results demonstrate Cr(VI) adsorption on PPGA is a spontaneous endothermic process. The adsorption mechanism involves electrostatic attraction, redox, and chelation. In general, this work provides a simple and eco-friendly way to prepare PPGA, which has a great potential for actual application of Cr(VI) removal in effluent.

Structural Investigation of DHICA Eumelanin Using Density Functional Theory and Classical Molecular Dynamics Simulations.

Eumelanin is an important pigment, for example, in skin, hair, eyes, and the inner ear. It is a highly heterogeneous polymer with 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and 5,6-dihydroxyindole (DHI) building blocks, of which DHICA is reported as the more abundant in natural eumelanin. The DHICA-eumelanin protomolecule consists of three building blocks, indole-2-carboxylic acid-5,6-quinone (ICAQ), DHICA and pyrrole-2,3,5-tricarboxylic acid (PTCA). Here, we focus on the self-assembly of DHICA-eumelanin using multi-microsecond molecular dynamics (MD) simulations at various concentrations in aqueous solutions. The molecule was first parameterized using density functional theory (DFT) calculations. Three types of systems were studied: (1) uncharged DHICA-eumelanin, (2) charged DHICA-eumelanin corresponding to physiological pH, and (3) a binary mixture of both of the above protomolecules. In the case of uncharged DHICA-eumelanin, spontaneous aggregation occurred and water molecules were present inside the aggregates. In the systems corresponding to physiological pH, all the carboxyl groups are negatively charged and the DHICA-eumelanin model has a net charge of -4. The effect of K+ ions as counterions was investigated. The results show high probability of binding to the deprotonated oxygens of the carboxylate anions in the PTCA moiety. Furthermore, the K+ counterions increased the solubility of DHICA-eumelanin in its charged form. A possible explanation is that the charged protomolecules favor binding to the K+ ions rather than aggregating and binding to other protomolecules. The binary mixtures show aggregation of uncharged DHICA-eumelanins; unlike the charged systems with no aggregation, a few charged DHICA-eumelanins are present on the surface of the uncharged aggregation, binding to the K+ ions.

Feature MS fragments-based method for identification of toxic furanoids in biological samples.

Numerous furan-containing compounds have been reported to be toxic. The toxicity may be attributed to the metabolic activation of the furan ring to cis-enediones. Identification of unknown furans that undergo bioactivation is challenging. Here, we present a novel approach that enables non-targeted profiling of bioactivation of unknown furanoids both in vitro and in vivo. Cyclic pyrrole-glutathione conjugate was the predominant product of cis-enediones with glutathione. The shared glutathione substructure of conjugates was capable of generating four constant and signature fragments under collision-induced dissociation (CID) in the mass spectrometer, including neutral loss fragments 103.0269 Da and 146.0691 Da and product ions at m/z 130.0499 and 177.0328. The unique structure and high abundance of conjugates in combination with the consistency and specificity of CID fragmentation brought extraordinarily high selectivity and reliability for the four fragments as a fingerprint of bioactivated furanoids. The bioactivated furanoids can be identified by screening the four fragments in high-resolution MS/MS datasets using the neutral loss filtering and diagnostic fragmentation filtering of data post-acquisition software MZmine. The simultaneous formation of four individual signal points in the filtering channel with the same precursor ion and retention time was assigned to be furanoids. The method has been rigorously validated. In the pooled urine samples from nine model furanoids-treated mice, nine cis-enediones from the parent furanoids and two from furanoid metabolites were accurately detected and identified. The method showed great performance in non-targeted profiling bioactivated furanoids and their metabolites in urine samples of herbal extract-treated mice.

In-Situ Intercalating of Silica Nanospheres into Polypyrrole During Its Electropolymerization to Prepare a Sorbent for Headspace Microextraction of Aldehydes in Edible Oils.

The quality of edible oils is significantly affected by autoxidation of lipids, which alters their flavor and nutritional quality through production of toxic materials like aldehydes (an important class of oil deterioration markers). Herein, an amino-silica nanospheres/polypyrrole (ASNS/PPy) nanocomposite sorbent was synthesized and used as the fiber coating for headspace solid-phase microextraction (HS-SPME) of aldehydes in edible oils, followed by gas chromatography (GC) separation and determination. Amino-silica nanoparticles were prepared by an amended Stöber method and composited with polypyrrole during its electropolymerization on the surface of a platinized stainless-steel fiber. The synergy between in-situ electropolymerization and rough surface of the platinized metal substrate created a durable fiber coating with unique uniformity, cohesiveness, and adsorption properties. The synthesized nanocomposite was characterized using Fourier transform infrared spectroscopy and scanning electron microscopy techniques. The performance of the prepared fiber was optimized by investigating the affecting variables including extraction temperature and time, stirring rate, and desorption conditions. The obtained limits of detection for hexanal and heptanal in sunflower oil were 0.005-0.009 µg mL-1. The prepared fiber exhibited excellent repeatability and reproducibility with the intra-fiber and inter-fiber relative standard deviations in the ranges of 3.9-8.8% and 7.3-15.1%, respectively. The proposed HS-SPME-GC strategy was successfully applied for the analysis of aldehydes in commercial edible oil samples.

Determination of 2-Acetyl-1-pyrroline via a Color-Change Reaction Using Chromium Hexacarbonyl.

At present, there is no colorimetric method for the quantitation of the aroma compound 2-acetyl-1-pyrroline (2AP). A novel colorimetric method was developed for the determination of 2AP content using chromium hexacarbonyl (Cr(CO)6) as a reagent. The reaction of synthetic 2AP with chromium hexacarbonyl reagent solution in the presence of light produced a green product with an absorption maximum (λmax) at 623 nm. GC-MS was used to confirm the color-change reaction, which showed the loss of 2AP after the addition of Cr(CO)6. This novel method enables facile and cost-effective determination of 2AP in fragrant rice. A comparative analysis of fragrant and nonfragrant rice grain extracts showed that no color-change reaction occurred with the nonfragrant rice sample. A limit of detection (LOD) of 2.00 mg L-1 was determined by method validation with an effective linear concentration ranging from 5.00 to 60.00 mg L-1 of 2AP. The results obtained using the developed colorimetric method were consistent with those obtained by automated static headspace gas chromatography with nitrogen-phosphorus detection (SHS-GC-NPD).

Formation of Volatile Heterocyclic Compounds and Open-Chain Amides of Theanine in Model Systems with Glucose, Tea Leaves, and Tea Extract under Tea-Roasting Conditions.

Theanine is a non-proteinogenic amino acid found in the tea plant Camellia sinensis. At an elevated temperature (>90 °C), it released two major volatile compounds 1-ethyl-1,5-dihydro-2H-pyrrol-2-one and N-ethylsuccinimide. Other products were identified, including 10 pyrroles and 12 amides/imides. The formation of the two major compounds was proposed to be initiated by the deamination of theanine and through the intermediate α-keto acid. In the presence of glucose, the two major products and many other volatiles from theanine thermal degradation were accelerated and further Maillard reactions occurred. A total of 56 compounds were identified in the model system of theanine and glucose, including 12 amides/imides, 16 pyrazines, 16 pyrroles and other N-heterocycles, and 12 furans and other O-heterocycles. Although most of the reaction products were detected in tea leaves and in their aqueous extract with or without the addition of theanine under the same experiment conditions, imides and amides were considerably suppressed, left only minute amounts, or were even no longer detectable. Pyrazines and pyrroles were also shown at reduced concentrations as a result of the interaction with tea components but to a lesser extent. A total of 16 and 12 pyrazines were identified in the theanine/glucose reaction system and tea leaves/aqueous extract after roasting, respectively. The results indicated that pyrazines and other main volatiles in roasted tea leaves were formed from the Maillard reactions of the aqueous fraction of tea leaves. Theanine participated in the formation of pyrazines in tea leaves under roasting conditions.

Insight into aroma dynamic changes during the whole manufacturing process of chestnut-like aroma green tea by combining GC-E-Nose, GC-IMS, and GC × GC-TOFMS.

Processing is the crucial factor for green tea aroma quality. In this study, the aroma dynamic changes throughout the manufacturing process of chestnut-like aroma green tea were investigated with gas chromatography electronic nose (GC-E-Nose), gas chromatography-ion mobility spectrometry (GC-IMS), and comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC-TOFMS). GC-IMS identified 33 volatile compounds while GC × GC-TOFMS identified 211 volatile components. Drying exerted the greatest influence on the volatile components of chestnut-like aroma green tea, and promoted the generation of heterocyclic compounds and sulfur compounds which were commonly generated via the Maillard reaction during the roasting stage. A large number of heterocyclic compounds such as 1-methyl-1H-pyrrole, pyrrole, methylpyrazine, furfural, 2-ethyl-5-methylpyrazine, 1-ethyl-1H-pyrrole-2-carboxaldehyde, and 3-acetylpyrrole were newly formed during the drying process. This study also validated the suitability of GC-E-Nose combined with GC-IMS and GC × GC-TOFMS for tracking the changes in volatile components of green tea throughout the manufacturing process.

Enhanced sorption and reduction of Cr(VI) by the flowerlike nanocomposites combined with molybdenum disulphide and polypyrrole.

Developing high-performance adsorbent for hexavalent chromium (Cr(VI)) elimination presents an enticing prospect in environmental remediation. Herein, three-dimensional flowerlike nanospheres composed of molybdenum disulphide and polypyrrole (MoS2@PPy) were successfully prepared via a one-pot hydrothermal and subsequent carbothermal reduction process for the removal of Cr(VI). The effects of pH, adsorbent dosage, co-existing ions, initial Cr(VI) concentration and temperature were investigated systematically by batch experiments. Benefiting from the incorporation of MoS2, the obtained MoS2@PPy composites showed a dramatic increase of specific surface area (149.82 m2·g-1) and adsorption capacity (230.97 mg·g-1) when compared with the pure PPy nanoparticles. Based on the thermodynamics study and X-ray photoelectron spectroscopy analyses, the removal process of Cr(VI) was proved to be exothermic and spontaneous, and accessible under-coordinated Mo(IV) and pyrrolic N groups coupled with redox reactions were conducive to the efficient removal of Cr(VI). Attractively, the MoS2@PPy acted as the electron donor could also activate peroxymonosulphate for the efficient degradation of organic contaminants. These results suggested that the MoS2@PPy was promising in Cr(VI) elimination and other kinds of organic pollutants removal in wastewater.

[18F]Nifene PET/CT Imaging in Mice: Improved Methods and Preliminary Studies of α4ß2* Nicotinic Acetylcholinergic Receptors in Transgenic A53T Mouse Model of α-Synucleinopathy and Post-Mortem Human Parkinson's Disease.

We report [18F]nifene binding to α4ß2* nicotinic acetylcholinergic receptors (nAChRs) in Parkinson's disease (PD). The study used transgenic Hualpha-Syn(A53T) PD mouse model of α-synucleinopathy for PET/CT studies in vivo and autoradiography in vitro. Additionally, postmortem human PD brain sections comprising of anterior cingulate were used in vitro to assess translation to human studies. Because the small size of mice brain poses challenges for PET imaging, improved methods for radiosynthesis of [18F]nifene and simplified PET/CT procedures in mice were developed by comparing intravenous (IV) and intraperitoneal (IP) administered [18F]nifene. An optimal PET/CT imaging time of 30-60 min post injection of [18F]nifene was established to provide thalamus to cerebellum ratio of 2.5 (with IV) and 2 (with IP). Transgenic Hualpha-Syn(A53T) mice brain slices exhibited 20-35% decrease while in vivo a 20-30% decrease of [18F]nifene was observed. Lewy bodies and α-synuclein aggregates were confirmed in human PD brain sections which lowered the [18F]nifene binding by more than 50% in anterior cingulate. Thus [18F]nifene offers a valuable tool for PET imaging studies of PD.

Evaluation of Dissipation Behavior, Residues, and Dietary Risk Assessment of Fludioxonil in Cherry via QuEChERS Using HPLC-MS/MS Technique.

The chemical fungicide fludioxonil is widely used to control post-harvest fungal disease in cherries. This study was implemented to investigate the dissipation behaviours and residues of fludioxonil on cherries. A reliable and efficient analytical method was established. Cherry samples from four product areas were analyzed by QuEChERS and HPLC-MS/MS methods with acceptable linearity (R2 > 0.99), accuracy (recoveries of 81-94%), and precision (relative standard deviation of 2.5-11.9%). The limits of quantification (LOQs) and limits of detection (LODs) of cherries were 0.01 mg/kg and 0.005 mg/kg. The dissipation of fludioxonil on cherries followed first order kinetics with half-lives of 33.7-44.7 days. The terminal residues of fludioxonil were all lower than 5.00 mg/kg, which is the MRL recommended by the European Commission. According to Chinese dietary patterns and terminal residue distributions, the risk quotient (RQs) of fludioxonil was 0.61%, revealing that the evaluated cherries exhibited an acceptably low dietary risk to consumers.

The Enantiomeric Discrimination of 5-Hexyl-2-methyl-3,4-dihydro-2H-pyrrole by Sulfobutyl ether-ß-cyclodextrin: A Case Study.

1-Pyrrolines are important intermediates of active natural products, such as the 2,5-dialkyl-1-pyrroline derivatives found in fire ant venoms. Here, 5-hexyl-2-methyl-3,4-dihydro-2H-pyrrole was synthesized by the enzymatic transamination/cyclization of 2,5-undecadione, and enantiodifferenciation was successfully achieved by capillary electrophoresis with sulfobutyl ether-ß-cyclodextrin as the chiral selector. The rationale of the enantiomeric discrimination was based on the results of a docking simulation that revealed the higher affinity of (S)-5-hexyl-2-methyl-3,4-dihydro-2H-pyrrole for the sulfobutyl ether-ß-cyclodextrin.

Effect of rainfall and altitude on the 2-acetyl-1-pyrroline and volatile compounds profile of black glutinous rice (Thai upland rice).

BACKGROUND: The aroma intensity of rice is mixed of more than 100 volatile compounds. Fragrant rice is famous in Thailand due to specific aroma. One important volatile compound that contributes to aromatic intensity and represents a characteristic compound for fragrant rice is 2-acetyl-1-pyrroline (2AP). The production of volatile compounds and 2AP in rice is affected by various environmental factors, such as cultivation area, water content and temperature. RESULTS: Our research employed 29 samples of fragrant colored rice (Luempua black glutinous rice cultivar) from Phetchabun province, Thailand from three harvested rice seasons (2015-2017). The cultivation areas of rice samples exhibited differences in rainfall (976.6-1260.5 mm y r-1 ) and altitude (26.26-1033.41 m). All volatile compounds and 2AP were identified by gas chromatography-mass spectrometry (GC-MS). In 29 rice samples, 2AP levels decreased with increasing of rainfall. The increase in altitude seems to increase 2AP levels, but some high-altitude cultivation areas resulted in low 2AP content. A variety of volatile compounds was observed in 29 rice samples from different cultivation areas. The results showed that differences in altitude and rainfall affected the production of volatile compounds in fragrant colored rice. Moreover, our results indicated that different environmental conditions in cultivation areas potentially affect volatile compound production during rice growth. CONCLUSION: The increase in 2AP levels under low rainfall conditions and the assortment of volatile compound varieties produced in colored rice grown in different cultivation areas may be useful information for rice cultivation management striving to produce rice with high aromatic intensities. © 2021 Society of Chemical Industry.

Rapid, High-Throughput Quantitation of Odor-Active 2-Acetyl Azaheterocycles in Food Products by UHPLC-MS/MS.

A rapid, high-throughput method for the quantitation of the 2-acetyl azaheterocycles, 2-acetyl-1-pyrroline, 2-acetyl-1,4,5,6-tetrahydropyridine, 2-acetylpyrazine, and 2-acetyl-2-thiazoline, in different food products, by liquid chromatography-tandem mass spectrometry (LC-MS/MS), was developed. The quick extraction by bead beater homogenization, fast derivatization by 3-nitrophenylhydrazine (40 °C, 2 h), and efficient LC separation make this method suitable for high-throughput analysis. As established in this study, the highly precise LC-MS/MS method applies to different food products or beverages without requiring further adjustment. The analysis was performed with sample amounts of 0.2-0.5 g, and limit of quantitation values of 0.6, 0.5, 0.6, and 1.0 µg/kg were obtained for 2-acetyl-1-pyrroline, 2-acetyl-1,4,5,6-tetrahydropyridine, 2-acetylpyrazine, and 2-acetyl-2-thiazoline, respectively. Thus, it was possible to quantitate the analytes in the range of their odor thresholds.

Comparison of the sensory properties of fragrant and non-fragrant rice (Oryza sativa), focusing on the role of the popcorn-like aroma compound 2-acetyl-1-pyrroline.

2-Acetyl-1-pyrroline (2-AP) has been widely reported as a key contributor to the popcorn-like aroma of fragrant rice (Oryza sativa). To gain a greater understanding of its contribution to the aroma in both fragrant and non-fragrant rice, sensory profiling was conducted with a trained panel to examine the sensory properties of six boiled rice samples, three fragrant and three non-fragrant varieties. The intensity of the popcorn note as an orthonasal odour, a retronasal flavour and as an after-effect was significantly higher in fragrant rice than in non-fragrant rice. However, panellists could not differentiate these popcorn attributes between the three different fragrant rice varieties. 2-AP was extracted from the boiled rice samples by headspace solid-phase microextraction and quantified by gas chromatography-mass spectrometry. 2-AP was below the limits of quantitation in non-fragrant varieties; however, gas chromatography-olfactometry of samples indicated the presence of 2-AP in both raw fragrant and non-fragrant rice varieties.

Determination of exogenous prohibited flavour compounds added in coffee using gas chromatography triple quadrupole tandem massspectrometry and gas chromatography/combustion/isotope ratio mass spectrometry.

An analytical method based on gas chromatography coupled to triple quadrupole tandem mass spectrometry (GC-MS/MS) was developed for the simultaneous determination of exogenous prohibited flavour compounds in coffee samples. In addition, gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) was developed to determine the origin of the founded prohibited flavour compound, N-methylpyrrole-2-carboxaldehyde (NMPCA). The good selectivity and sensitivity achieved in multiple reactions monitoring (MRM) mode allowed satisfactory confirmation and quantitation for the flavour compounds. The limits of detection (LODs) and limits of quantitation (LOQs) of these compounds were in the range of 0.0005-5.0 µg/kg and 0.002-16.0 µg/kg, respectively. The coffee samples were extracted with simultaneous distillation extraction (SDE) and NMPCA was analysed on a GC/C/IRMS system. The δ13C values of endogenous NMPCA in coffee beans were within a range of -35.0‰ to -31.1‰, whereas exogenous NMPCA was the range from -27.9‰ to -23.9‰. The validation results revealed that the GC-MS/MS method was sensitive and reliable, and the origin of NMPCA can be distinguished by GC/C/IRMS. Finally, this method was successfully applied to coffee samples analysis and NMPCA was found in coffee samples.

Secondary Metabolites of an of Streptomyces griseorubens Isolate Are Predominantly Pyrrole- and Linoleic-acid like Compounds.

The study involved the isolation and identification of a member of Streptomyces griseorubens and the identification of its secondary metabolite content. Two extract samples were prepared by using butanol and chloroform. In the analyses of the extracts TLC, FT-IR, and GC-MS were employed. Butanol extract appeared to be dominated by three different pyrrole compounds (43.59%), while two fatty acids, linoleic- and erucic acids, were the most abundant secondary metabolites in the chloroform extract, 27.57% and 12.34%, respectively. Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-compound was represented by a single and distinct band on the thin layer chromatography plate. In GC-MS spectra, it also constituted 13.50% of the butanol extract.

Non-rhizobial endophytic (NRE) yeasts assist nodulation of Rhizobium in root nodules of blackgram (Vigna mungo L.).

The signal orchestration between legumes and the rhizobia attribute to symbiotic nitrogen fixation through nodule formation. Root nodules serve as a nutrient-rich reservoir and harbor diverse microbial communities. However, the existence of non-rhizobial endophytes (NRE) and their role inside the root nodules are being explored; there is no evidence on yeast microflora inhabiting nodule niche. This study focused on unraveling the presence of yeast in the root nodules and their possible function in either nodulation or signal exchange. From the root nodules of blackgram, two yeast strains were isolated and identified as Candida glabrata VYP1 and Candida tropicalis VYW1 based on 18S rRNA gene sequencing and phylogeny. These strains possessed plant growth-promoting traits viz., IAA, ACC deaminase, siderophore, ammonia, and polyamine production. The functional capacity of endophytic yeast strains, and their interaction with Rhizobium sp. was further unveiled via profiling volatile organic compounds (VOC). Among the VOCs, α-glucopyranoside and pyrroloquinoline pitches a pivotal role in activating lectin pathways and phosphorous metabolism. Further, lectin pathways are crucial for nodulating bacterium, and our study showed that these endophytic yeasts assist nodulation by Rhizobium sp. via activating the nod factors. The plant growth-promoting traits of NRE yeast strains coupled with their metabolite production, could recruit them as potential drivers in the plant-microbe interaction.