Rice foliar-adapted Pantoea species: Promising microbial biostimulants enhancing rice resilience against foliar pathogens, Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae.

Foliar fungal blast and bacterial leaf blight have significant impacts on rice production, and their management through host resistance and agrochemicals has proven inadequate. To achieve their sustainable management, innovative approaches like leveraging the foliar microbiome, which collaborates with plants and competes against pathogens, are essential. In our study, we isolated three Pantoea strains (P. agglomerans Os-Ep-PPA-1b, P. vagans Os-Ep-PPA-3b, and P. deleyi Os-Ep-VPA-9a) from the rice phylloplane. These isolates exhibited antimicrobial action through their metabolome and volatilome, while also promoting rice growth. Our analysis, using Gas Chromatography-Mass Spectrometry (GC-MS), revealed the presence of various antimicrobial compounds such as esters and fatty acids produced by these Pantoea isolates. Inoculating rice seedlings with P. agglomerans and P. vagans led to increased root and shoot growth. Additionally, bacterized seedlings displayed enhanced immunocompetence, as evidenced by upregulated expressions of defense genes (OsEDS1, OsFLS2, OsPDF2.2, OsACO4, OsICS OsPR1a, OsNPR1.3, OsPAD4, OsCERK1.1), along with heightened activities of defense enzymes like Polyphenol Oxidase and Peroxidase. These plants also exhibited elevated levels of total phenols. In field trials, the Pantoea isolates contributed to improved plant growth, exemplified by increased flag-leaf length, panicle number, and grains per panicle, while simultaneously reducing the incidence of chaffy grains. Hypersensitivity assays performed on a model plant, tobacco, confirmed the non-pathogenic nature of these Pantoea isolates. In summary, our study underscores the potential of Pantoea bacteria in combatting rice foliar diseases. Coupled with their remarkable growth-promoting and biostimulant capabilities, these findings position Pantoea as promising agents for enhancing rice cultivation.

Metabolomic profiling of virulent and non-virulent Beauveria bassiana strains: insights into the pathogenicity of Tetranychus truncatus.

In this study, the pathogenicity of local Beauveria bassiana strains was elucidated using molecular and metabolomics methodologies. Molecular verification of the B. bassiana-specific chitinase gene was achieved via phylogenetic analysis of the Bbchit1 region. Subsequent metabolomic analyses employing UPLC-Q-TOF-MS revealed a different number of non-volatile metabolite profiles among the six B. bassiana strains. Bb6 produced the most non-volatile compounds (17) out of a total of 18, followed by Bb15 (16) and Bb12 (15). Similarly, Bb5, Bb8, and Bb21, three non-virulent B. bassiana strains, produced 13, 14, and 14 metabolites, respectively. But unique secondary metabolites like bassianolide and beauvericin, pivotal for virulence and mite management, were exclusively found in the virulent strains (Bb6, Bb12, and Bb15) of B. bassiana. The distinctive non-volatile metabolomic profiles of these strains underscore their pathogenicity against Tetranychus truncatus, suggesting their promise in bio-control applications.

Differential regulation of glucosinolate-myrosinase mediated defense determines host-aphid interaction in Indian mustard Brassica juncea L.

BACKGROUND: India's oilseed economy falls short of self-sufficiency and is supplemented by huge imports every year. Increasing national productivity of the major oilseeds is confronted with yield losses due to diverse biotic and abiotic stresses. The productivity of Indian mustard (Brassica juncea Linnaeus), belonging to the family Brassicaceae, is significantly reduced due to damage caused by mustard aphids (Lipaphis erysimi Kaltenbach, Hemiptera: Aphididae). Rapid colonization by the nymphs makes it difficult to protect the crop through agrochemicals. Aphids release effector molecules to modulate the host-defence responses. Glucosinolates (GSLs) extensively found in Brassicaceae family, are hydrolysed by myrosinase into toxic compounds that deter herbivore insects. METHODS: Here, we investigated the differential activation of the glucosinolate-myrosinase pathway in mustard manifesting susceptibility and resistance to different aphid species. Mustard plants were challenged by two different aphid species mustard aphid and cowpea aphid (Aphis craccivora Koch, Hemiptera: Aphididae) leading to complete host-susceptibility in one case and resistance in the other, respectively. Differential regulation of the GSL biosynthetic pathway and myrosinase activity was assessed by gene expression study and ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC- QToF-ESL-MS). RESULTS: Gene expression study identified selective transcriptional attenuation of the key GSL biosynthetic and myrosinase gene in mustard when challenged with mustard aphid. In contrary, the activation of GSL biosynthetic genes in conjunction with myrosinase at the transcriptional level was profound in mustard, when challenged with cowpea aphid. UPLC-MS analysis showed higher turnover in the hydrolysis of glucosinolates by myrosinase which led to concomitant generation of glucose as byproduct in response to cowpea aphid in mustard plants. CONCLUSION: GSL-myrosinase pathway is specifically attenuated by the successful aphid species in mustard and thus plays a pivotal role in determining the outcome of the B. juncea-aphid interaction. The results open up a new genetic modification strategy for developing resistance against aphids.

Hydroxamate siderophores secreted by plant endophytic Pseudomonas putida elicit defense against blast disease in rice incited by Magnaporthe oryzae.

Our study focuses on hydroxamate-type siderophores from Pseudomonas putida BP25, known for chelating ferric iron and aiding microbial growth in iron-deficient environments. Confirmed through CAS-agar and tetrazolium tests, a purified siderophore extract was obtained via ion-exchange chromatography. Applying varying concentrations of this siderophore to rice seedlings demonstrated concentration-dependent effects on shoot and root phenotypes. Prophylactic application on rice leaves significantly reduced blast severity (68.7%-97.0%), surpassing curative application (47.5%-86.87%). Additionally, the siderophore treatment elevated peroxidase, polyphenol oxidase, and total phenols in rice plants. Defense-related genes linked to salicylic acid (OsPR1.1, OsNPR1, and OsPDF2.2), and other pathways (Oshox24, OsCLE, and OsGLP3-3, OsEIN2.4, and OsCSE) promoting blast suppression showed upregulation. However, the OsACS6 gene associated with ethylene-induced internodal elongation was significantly downregulated. Overall, our findings propose that the siderophore from P. putida BP25 induces defense gene transcription, offering potential for sustainable rice production via bio-formulation.

Differential effects of rhizobacteria from uninfected and infected tomato on Meloidogyne incognita under protected cultivation.

Intermingled uninfected and root-knot nematode-infected tomato plants are commonly observed under protected cultivation. To understand the role of rhizobacteria underlying the susceptibility to nematode infectivity in these tomato plants, 36 rhizobacteria (18 from each type) with morphologically distinct colony characteristics were isolated from the rhizosphere of uninfected and root-knot nematode-infected tomato plants. The in vitro nematicidal potential of rhizobacteria from the uninfected rhizosphere was significantly higher than that from the infested rhizosphere. The three most effective antagonists were identified as Microbacterium laevaniformans, Staphylococcus kloosii, Priestia aryabhattai from root-knot-nematode-infected tomato rhizosphere and Staphylococcus sciuri, Bacillus pumilus, and Priestia megaterium from the rhizosphere of uninfected tomato. Volatile organic compounds from these rhizobacteria were characterized. Except for S. kloosi, the soil drenching with other rhizobacteria significantly reduced juvenile penetration (>60%) in tomato roots. Furthermore, the application of a single or consortium of these rhizobacteria affected nematode reproduction in tomato. Four consortia of rhizobacteria (S. sciuri + B. pumilus + P. megaterium), (B. pumilus + P. megaterium), (S. sciuri + B. pumilus), and (S. sciuri + P. megaterium) from uninfested rhizosphere and two consortia (M. laevaniformans + P. aryabhattai), (M. laevaniformans + S. kloosii + P. aryabhattai) from infested rhizosphere (IRh) effectively reduced M. incognita reproduction and considerably enhanced plant growth and yield in tomato. The nematicidal efficacy, however, decreased when S. kloosii was applied in the consortium. These distinctive effects illustrate how the plant susceptibility to nematode infectivity is modulated under natural conditions.

Nematicidal and Molecular Docking Investigation of Essential Oils from Pogostemon cablin Ecotypes against Meloidogyne incognita.

Root-knot nematode, Meloidogyne incognita is one of the most destructive nematodes worldwide. Essential oils (EOs) are being extensively utilized as eco-benign bionematicides, although the precise mechanism of action remains unclear. Pogostemon cablin Benth. is well-known as "Patchouli". It is native to South East Asia and known for ethno-pharmacological properties. In this study, chemical composition and potential nematicidal effect of EOs hydrodistilled from the leaves of P. cablin grown at three different locations in India were comprehensively investigated to correlate their mechanism of action for target specific binding affinities toward nematode proteins. Aromatic volatile Pogostemon essential oils (PEO) from Northern India (PEO-NI), Southern India (PEO-SI) and North Eastern India (PEO-NEI) were analyzed by Gas Chromatography-Mass Spectrometry (GC/MS) to characterize forty volatile compounds. Maximum thirty-three components were identified in PEO-NEI. Sesquiterpenes were predominant with higher content of α-guaiene (2.3-24.4 %), patchoulol (6.1-32.7 %) and α-bulnesene (5.9-27.1 %). Patchoulol was the major component in PEO-SI (32.7±1.2 %) and PEO-NEI (29.2±1.1 %), while α-guaiene in PEO-NI (24.4±1.2 %). In vitro nematicidal assay revealed significant nematicidal action (LC50 44.6-87.0 µg mL-1 ) against juveniles of M. incognita within 24 h exposure. Mortality increases with increasing time to 48 h (LC50 33.6-71.6 µg mL-1 ) and 72 h (LC50 27.7-61.2 µg mL-1 ). Molecular modelling and in silico studies revealed multi-modal inhibitive action of α-bulnesene (-22 to -13 kJ mol-1 ) and α-guaiene (-22 to -12 kJ mol-1 ) against three target proteins namely, acetyl cholinesterase (AChE), odorant response gene-1 (ODR1), odorant response gene-3 (ODR3). Most preferable binding mechanism was observed against AChE due to pi-alkyl, pi-sigma, and hydrophobic interactions. Structure nematicidal activity relationship suggested the presence of hydroxy group for nematicidal activity is nonessential, rather highly depends on synergistic composition of sesquiterpene hydrocarbons.

Brassica nigra essential oil: In-vitro and in-silico antibacterial efficacy against plant pathogenic and nitrifying bacteria.

The present study was aimed to examine the antibacterial potential of Brassica nigra essential oil (BNEO) against Ralstonia solanacearum, causal agent of bacterial wilt and Nitrosomonas sp., the nitrifying bacteria. In poisoned food assay, BNEO showed 100% growth inhibition of R. solancearum at ≥ 125 µg mL-1. Revalidation of findings by volatile assay employing inverted Petri plate technique exhibited 100% bacterial growth inhibition caused by vapors of BNEO, even at 50 µg mL-1 concentration. In the broth microdilution assay, the BNEO exhibited significant antibacterial activity only at higher concentrations (>500 µg mL-1). At 500 µg mL-1, BNEO showed 80% bacterial growth inhibition over control, which was at par with that of streptomycin (5 µg mL-1). In resazurin microtitre-plate assay, the maximum concentration of BNEO, at which color change occurred was 512 µg mL-1 (T9), and thus 512 µg mL-1 was concluded as the minimum inhibitory concentration (MIC). BNEO effectively inhibited the activity of Nitrosomonas spp. with 30-65% nitrification inhibition at the dose of 400 mkg-1 of Urea-N. Homology modeled protein targets assisted computational tool-based novel analysis helped to understand that the antibacterial potency of BNEO is due to preferable binding efficiency of allyl isothiocyanate (AITC), the major active ingredient of BNEO.

Chemometric approach based characterization and selection of mid-early cauliflower for bioactive compounds and antioxidant activity.

The present study was aimed to analyse bioactive compounds (total phenolics, ascorbic acid and sinigrin) and antioxidant activity in 14 mid-early cauliflower genotypes. Significant differences (pb 0.05) were observed among the genotypes for all bioactive compounds and antioxidant activity. Total phenolics content of curd were ranged from 20.36 to 48.93 mg gallic acid equivalent (GAE) 100 g-1 fresh weight (FW) which showed 2.5 times variation. The ascorbic acid content was maximum in DC522 (88.53 mg 100 g-1 FW) followed by Pusa Sharad (65.64 mg 100 g-1 FW) while minimum in DC310 (39.62 65.64 mg 100 g-1 FW). Wide variation was observed for cupric reducing antioxidant capacity and ferric reducing antioxidant power ranging from 9.04 to 20.83 mg GAE 100 g-1 FW and 13.11 to 26.31 mg GAE 100 g-1 FW, respectively. Sinigrin was found to be highest in DC306 (39.50 µmol 100 g-1 FW) for leaf and in DC326 (36.93 µmol 100 g-1 FW) for curd sample. The cauliflower genotypes were classified based on chemometric approaches namely principal component analysis (PCA) and agglomerative hierarchical clustering (AHC). The first two principal components (PC1 and PC2) explained 50.62% and 23.28% of total variance, respectively. The AHC as revealed by heat map classified cauliflower genotypes into four main groups based on measured traits. The information is useful for developing varieties and/or hybrids rich in bioactive compounds and antioxidant activity.

Valorisation of raw mango pickle industry waste into antimicrobial agent against postharvest fungal pathogens.

In mango pickle industry, a significant quantity of mango seed kernels is discarded as solid wastes. These seed kernels can be an ideal source for obtaining extracts rich in bioactive polyphenolic compounds with good antioxidant properties. The potential of mango kernel phenolic extract (MKPE) was investigated as a natural and effective antimicrobial agent for controlling major postharvest fungal pathogen infections, a significant threat to global food supply chains. Fungal pathogens contribute to the deterioration of fruits, vegetables, and grains during storage and transportation, leading to economic losses and compromised food safety. MKPE was obtained from pickling variety 'Ramkela' raw mango kernels, and its phenolic composition was characterized using LC-MS. The in vitro antifungal activity of MKPE against Botrytis cinerea, Colletotrichum gloeosporoides, and Rhizopus stolonifer was evaluated in vitro. A concentration-dependent inhibition of fungal radial growth against all three pathogens was observed, exhibiting the potential of MKPE as a valuable natural resource for addressing postharvest losses caused by fungal pathogens. The extraction process yielded a total phenolic content of 2128 mg GAE/100 g. Major polyphenolic bioactive compounds present were mangiferin, quercetin, and rhamnetin. The in-vitro antimicrobial assay showed reduction in the radial growth and inhibition percent of the pathogens. EC50 values of MKPE for B. cineria, C. gloeosporoides, and R. stolonifer was found to 364.17, 963.8 and 926 ppm, respectively. Our results demonstrate an economical, sustainable, and eco-friendly approach to manage post-harvest diseases rendered by fungi using mango MKPE from pickling industry waste.

Lippia alba-a potential bioresource for the management of Spodoptera frugiperda (Lepidoptera: Noctuidae).

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), a threat to maize production systems, is a polyphagous pest of global significance. There is no registered bioinsecticide of botanical origin to provide green remedy against this pest of concern. The present study reports for the first time the potency of the polar and non-polar bioinsecticidal leads sourced from Lippia alba (Mill.) N.E. Br. leaves. Shade-dried leaves of L. alba were extracted and evaluated; based on preliminary bioassay, the ethyl acetate leaf extract of L. alba (LEAE) was found to be the most potent against FAW in the in vitro and in vivo studies. Ultraperformance liquid chromatography-quadrupole time-of-flight-mass spectrometric (UPLC-QToF-MS) analysis of LEAE revealed the rich chemical profile of 28 compounds, dominated by flavones, namely, naringenin, trihydroxy-dimethoxy flavone, and dihydroxy-trimethoxy flavone. Among others, glycosides, such as clerodendrin, calceolarioside E, forsythoside B, geniposide, and martynoside, and glucuronides, such as luteolin-7-diglucuronide, tricin-7-O-glucuronide, and luteolin-7-O-glucuronide, were also identified. LEAE exhibited exceptionally high in vitro [LC50 = 6,900 parts per million (ppm)] and in vivo (computed as damage score on a scale of 1-9) insecticidal activity against S. frugiperda, with no phytotoxicity at a dose as high as 20 times of LC50. LEAE also exhibited significant antifeedant, ovicidal, and growth regulatory activity at the 70-16,000 ppm (w/v) concentration range. In silico assessment revealed strong binding of martynoside, calceolarioside E, and forsythoside B with acetylcholinesterase-, sodium-, and chloride-dependent γ-aminobutyric acid (GABA) receptor and ryanodine receptor, respectively, facilitated by hydrogen bonds (conventional and C-H bonds) stabilized by hydrophobic pi-sigma, pi-pi stacked, pi-alkyl, and alkyl interactions. The present study established L. alba as a potential bioresource and secondary metabolite enriched LEAE as bioinsecticide for further product development.

Exploring protein structural adaptations and polyphenol interactions: Influences on digestibility in pigeon pea dal and whole grains under heat and germination conditions.

Pigeon pea, a protein-rich legume with low protein digestibility (PD) due to its high polyphenol content and other antinutritional factors (ANFs). Consequently, processing methods are crucial to improve PD. We investigated the effects of thermal treatments (cooking, hydrothermal, autoclaving, infrared rays) treatments and germination on modulation of PD, its properties and association with ANFs in two distinct genotypes based on polyphenol content: high (Pusa Arhar 2018-4) and low (ICP-1452). Treatments improved in vitro PD and essential amino acid content, with autoclaving showing significantly higher PD (ICP-1452: 90.4%, Pusa-Arhar 2018-4: 84.32%) ascribed to disruption of tight protein matrices. Significant increase in ß-turn, reduction in protein: starch, protein: polyphenol interactions as well as breakdown of storage proteins revealed by the analysis of protein structural properties. This study suggests thermal treatments, particularly autoclaving, can enhance pigeon pea protein's nutritional quality for its utilization as a new ingredient in development of healthy foods.

Effect of mobile phase on resolution of the isomers and homologues of tocopherols on a triacontyl stationary phase.

Reversed-phase liquid chromatographic (RPLC) separation of isomers and homologues of similar polarity is challenging. Tocopherol isomers and homologues are one such example. α, ß, γ, and δ-tocopherols have been successfully separated by RPLC on triacontyl (C30) stationary phase. System suitability was tested by using four mobile phases, and observed chromatographic separations of ß and γ-tocopherols were compared. Comparison indicated that methanol-tert-butyl methyl ether (TBME) 95:5 (v/v) at a flow rate of 0.75 mL min(-1) was the best mobile phase. Detection systems were also evaluated on the basis of limit of quantification; it was concluded that fluorescence detection was best. The method was validated by analysis of two homologues and two isomers of tocopherol in sesame, maize, and soybean samples. MS coupled with an ESI interface in negative-ion mode [M - H](-) was used for identification of individual components. It was concluded that addition of TBME to methanol was required to enhance the separation of ß and γ-tocopherols, although methanol alone provided similar results. The applicability of the method to cereal, pulse, and oilseed samples was confirmed. The reproducibility of the procedure was good, with relative standard deviations in the range 1.7-3.9%. Recovery of tocopherols added to sesame samples ranged from 91 to 99%.

Cadinene sesquiterpenes from Eupatorium adenophorum and their antifungal activity.

Bioactive constituents of Eupatorium adenophorum were investigated for antifungal activity. A structure-antifungal activity relationship of cadinene sesquiterpenes was predicted by evaluating individual derivatives. Cadinene derivatives were extracted from leaves of Eupatorium adenophorum using ethyl acetate. Five cadinene sesquiterpenes were isolated by column chromatography and Preparative Thin Layer Chromatography. Bioactivity of these cadinene sesquiterpenes were evaluated in vitro against four phytopathogenic fungi using poison food technique. Purified sesquiterpenes were spectroscopically elucidated as cadinan-3-ene-2,7-dione (1), 7-hydroxycadinan-3-ene-2-one (2), 5,6-dihydroxycadinan-3-ene-2,7-dione (3), cadinan-3,6-diene-2,7-dione (4) and 2-acetyl-cadinan-3,6-diene-7-one (5). Antifungal evaluation of these compounds against pathogenic fungi was found to be selective. Compound 1 was highly inhibitory towards S. rolfsii (ED50 181.60 ± 0.58 µgmL(-1)) and R. solani (ED50 189.74 ± 1.03 µgmL(-1)). Availability of plant material and significant antifungal activity makes the plant a potential source of antifungal agent and that can be exploited for the development of a natural fungicide.

Effect of thermal treatments on the matrix components, inherent glycemic potential, and bioaccessibility of phenolics and micronutrients in pearl millet rotis.

Pearl millet (PM) is a nutri-cereal rich in various macro and micronutrients required for a balanced diet. Its grains have a unique phenolic and micronutrient composition; however, the lower bioaccessibility of nutrients and rancidity of flour during storage are the major constraints in its consumption and wide popularity. Here, to explore the effect of different thermal processing methods, i.e., hydrothermal (HT), microwave (MW), and infrared (IR) treatments, on the digestion of starch, phenolics, and microelements (Fe and Zn), an in vitro digestion model consisting of oral, gastric and intestinal digestion was applied to PM rotis. The hydrothermally treated PM roti was promising as it showed lower inherent glycemic potential (60.4%) than the untreated sample (72.4%) and less enzymatic activities associated with rancidity in PM flour. FTIR revealed an increased ratio of 1047/1022 cm-1 in the hydrothermally treated sample, reflecting the enhancement of the structurally ordered degree and compactness of starch compared to other thermal treatments. A tighter and more compact microstructure with an agglomeration of starch in the hydrothermally treated PM flour was observed by SEM. These structural changes could provide a better understanding of the lower starch digestion rate in the hydrothermally treated flour. However, HT treatment significantly (P < 0.05) reduced the bioaccessibility of phenolics (10.6%) compared to native PM rotis and slightly reduced the Fe (2%) and Zn (3.2%) bioaccessibility present in PM rotis.

Exploring Potent Fungal Isolates from Sanitary Landfill Soil for In Vitro Degradation of Dibutyl Phthalate.

Di-n-butyl phthalate (DBP) is one of the most extensively used plasticizers for providing elasticity to plastics. Being potentially harmful to humans, investigating eco-benign options for its rapid degradation is imperative. Microbe-mediated DBP mineralization is well-recorded, but studies on the pollutant's fungal catabolism remain scarce. Thus, the present investigation was undertaken to exploit the fungal strains from toxic sanitary landfill soil for the degradation of DBP. The most efficient isolate, SDBP4, identified on a molecular basis as Aspergillus flavus, was able to mineralize 99.34% dibutyl phthalate (100 mg L-1) within 15 days of incubation. It was found that the high production of esterases by the fungal strain was responsible for the degradation. The strain also exhibited the highest biomass (1615.33 mg L-1) and total soluble protein (261.73 µg mL-1) production amongst other isolates. The DBP degradation pathway scheme was elucidated with the help of GC-MS-based characterizations that revealed the formation of intermediate metabolites such as benzyl-butyl phthalate (BBP), dimethyl-phthalate (DMP), di-iso-butyl-phthalate (DIBP) and phthalic acid (PA). This is the first report of DBP mineralization assisted with A. flavus, using it as a sole carbon source. SDBP4 will be further formulated to develop an eco-benign product for the bioremediation of DBP-contaminated toxic sanitary landfill soils.

Effect of Microbial Consortium Constructed with Lignolytic Ascomycetes Fungi on Degradation of Rice Stubble.

Microbial degradation is an effective, eco-friendly and sustainable approach for management of the rice residue. After harvesting a rice crop, removal of stubble from the ground is a challenging task, that forces the farmers to burn the residue in-situ. Therefore, accelerated degradation using an eco-friendly alternative is a necessity. White rot fungi are the most explored group of microbes for accelerated degradation of lignin but they are very slow in growth. The present investigation focuses on degradation of rice stubble using a fungal consortium constructed with highly sporulating ascomycetes fungi, namely, Aspergillus terreus, Aspergillus fumigatus and Alternaria spp. All three species were successful at colonizing the rice stubble. Periodical HPLC analysis of rice stubble alkali extracts revealed that incubation with ligninolytic consortium released various lignin degradation products such as vanillin, vanillic acid, coniferyl alcohol, syringic acid and ferulic acid. The efficiency of the consortium was further studied at different dosages on paddy straw. Maximum lignin degradation was observed when the consortium was applied at 15% volume by weight of rice stubble. Maximum activity of different lignolytic enzymes such as lignin peroxidase, laccase and total phenols was also found with the same treatment. FTIR analysis also supported the observed results. Hence, the presently developed consortium for degrading rice stubble was found to be effective in both laboratory and field conditions. The developed consortium or its oxidative enzymes can be used alone or combined with other commercial cellulolytic consortia to manage the accumulating rice stubble effectively.

New Insights on Endophytic Microbacterium-Assisted Blast Disease Suppression and Growth Promotion in Rice: Revelation by Polyphasic Functional Characterization and Transcriptomics.

Plant growth-promoting endophytic microbes have drawn the attention of researchers owing to their ability to confer fitness benefits in many plant species. Here, we report agriculturally beneficial traits of rice-leaf-adapted endophytic Microbacterium testaceum. Our polyphasic taxonomic investigations revealed its identity as M. testaceum. The bacterium displayed typical endophytism in rice leaves, indicated by the green fluorescence of GFP-tagged M. testaceum in confocal laser scanning microscopy. Furthermore, the bacterium showed mineral solubilization and production of IAA, ammonia, and hydrolytic enzymes. Tobacco leaf infiltration assay confirmed its non-pathogenic nature on plants. The bacterium showed antifungal activity on Magnaporthe oryzae, as exemplified by secreted and volatile organic metabolome-mediated mycelial growth inhibition. GC-MS analysis of the volatilome of M. testaceum indicated the abundance of antimicrobial compounds. Bacterization of rice seedlings showed phenotypic traits of MAMP-triggered immunity (MTI), over-expression of OsNPR1 and OsCERK, and the consequent blast suppressive activity. Strikingly, M. testaceum induced the transcriptional tradeoff between physiological growth and host defense pathways as indicated by up- and downregulated DEGs. Coupled with its plant probiotic features and the defense elicitation activity, the present study paves the way for developing Microbacterium testaceum-mediated bioformulation for sustainably managing rice blast disease.

Phylogenetic analysis, molecular characterization and virulence profiling based on toxoflavin gene of an Indian BG1 strain of Burkholderia glumae causing panicle blight of rice.

Bacterial panicle blight (BPB) caused by Burkholderia glumae (BG) has become significantly more prevalent in the rice-growing regions of North India. Based on virulence screening and in vitro quantification of toxoflavin, the BG strains were classified as hyper- (BG1 and BG3), moderate- (BG2, BG4, BG6, BG8, and BG9), and hypo- (BG5, BG7, and BG10) virulent. Plant inoculation assays with cell-free culture filtrate revealed strains with higher toxoflavin-producing ability had higher virulence. Based on 16S rRNA sequence, 6 isolates from Uttar Pradesh were grouped in clad C1; whereas, clad C2 exhibited 4 isolates, two each from Delhi and Uttar Pradesh. Strain BG1 being the most virulent Indian strain from Uttar Pradesh was further profiled for 11 tox genes. We found all the 11 tox genes present in strain BG1. In toxRABCDE cluster, all tox genes showed high similarity to B. glumae BGR1 except toxB, whereas in toxFGHIJ cluster toxF, toxG, toxH and toxI shared maximum similarity to B. glumae 336gr-1. tox genes of BG1 exhibited homology as well as divergence with B. gladioli. The domain prediction and protein association network analysis indicated the possible involvement of tox genes in the toxoflavin biosynthesis. As per our knowledge, this is the first report in India on characterization of tox genes cluster in B. glumae. Altogether, our study unravels a reliable method for identifying and characterizing B. glumae using tox genes and its relationship with disease production. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03660-6.

Metabolomic profiling and its association with the bio-efficacy of Aspergillus niger strain against Fusarium wilt of guava.

Bio-control agents are the best alternative to chemicals for the successful management of plant diseases. The fungus Aspergillus niger is known to produce diverse metabolites with antifungal activity, attracting researchers to exploit it as a bio-control agent for plant disease control. In the present study, 11 A. niger strains were isolated and screened for their antagonism against the guava wilt pathogen under in vitro and in planta conditions. Strains were identified morphologically and molecularly by sequencing the internal transcribed spacer (ITS), ß-tubulin, and calmodulin genes. The strains were evaluated through dual culture, volatile, and non-volatile methods under an in vitro study. AN-11, AN-6, and AN-2 inhibited the test pathogen Fusarium oxysporum f. sp. psidii (FOP) at 67.16%, 64.01%, and 60.48%, respectively. An in planta study was conducted under greenhouse conditions with 6 months old air-layered guava plants (var. Allahabad Safeda) by pre- and post-inoculation of FOP. The AN-11 strain was found to be effective under both pre- and post-inoculation trials. Furthermore, gas chromatography-mass spectrometry (GC-MS) analysis was carried out to characterize the volatile compounds of the most potential strain, A. niger. The hexane soluble fraction showed the appearance of characteristic peaks of hexadecenoic acid methyl ester (4.41%), 10-octadecanoic acid methyl ester (3.79%), dodecane (3.21%), undecane (3.19%), gibepyrone A (0.15%), 3-methylundecane (0.36%), and citroflex A (0.38%). The ethyl acetate fraction of the bio-control fungi revealed the occurrence of major antifungal compounds, such as acetic acid ethyl ester (17.32%), benzopyron-4-ol (12.17%), 1,2,6-hexanetriol (7.16%), 2-propenoic acid ethanediyl ester (2.95%), 1-(3-ethyloxiranyl)-ethenone (0.98%), 6-acetyl-8-methoxy dimethyl chromene (0.96%), 4-hexyl-2,5-dihydro dioxo furan acetic acid (0.19%), and octadecanoic acid (1.11%). Furthermore, bio-control abilities could be due to hyper-parasitism, the production of secondary metabolites, and competition for sites and nutrients. Indeed, the results will enrich the existing knowledge of metabolomic information and support perspectives on the bio-control mechanism of A. niger.

Macroporous resin-assisted enrichment, characterizations, antioxidant and anticandidal potential of phytochemicals from Trachyspermum ammi.

Extract of de-oiled seeds of Trachyspermum ammi was purified using macroporous resins and the performance of three resins were evaluated to enrich major phytochemical component. A HPLC method has been developed to separate major phytochemical constituents in the crude (CTAE) and partially purified extracts (PTAE). Macroporous resin assisted enrichment and purification suggested XAD-16 as the most efficient (yield 29.8%) followed by XAD-7HP and Diaion HP-20. Concentrated PTAE was subjected to multiple preparative-TLC to afford three compounds, namely, rosmarinic acid-3-O-glucopyranoside (TA-1), kaempferol-(coumaroyl-glucosyl)-rhamnoside (TA-2) and quercetin-3-O-galactoside (TA-3). The structure of these compounds was elucidated from their corresponding spectroscopic characterizations in FT-IR, HR-MS, and partially by 1 H NMR. Total phenolic and flavonoid contents of the extracts were determined. Antioxidant activity by DPPH and ABTS radical scavenging, CUPRAC assays indicated the highest antioxidant potential of CTAE. Among the compounds, TA-1 exhibited the highest scavenging activity in ABTS (IC50 33.41 µg/ml) and DPPH (IC50 69.23 µg/ml), however, relatively lower than CTAE. In vitro anti-candidal activity against virulent strains of Candida spp. revealed C. albicans 4718 as the most susceptible (23.9 µg/ml) to PTAE. PRACTICAL APPLICATIONS: Seeds of Trachyspermum ammi has been extensively investigated for volatile aromatic components of the essential oil. However, the de-oiled seeds have rarely been exploited for potential bioactive phenolics. The present investigation envisaged possible utilization of the de-oiled Trachyspermum seeds for its phenolic constituents, which could be used as natural antioxidant with additional benefits of anticandidal properties. Indeed, macroporous resin assisted enrichment and purification of extracts of T. ammi seeds generate valuable reference compounds, rosmarinic acid-3-O-glucopyranoside, kaempferol-(coumaroyl glucosyl)-rhamnoside, and quercetin-3-O-galactoside.