Indole Production Research Articles

First Report of Fire Blight Caused by Erwinia amylovora on Pear in Saudi Arabia.

In the spring of 2020 and 2021, pear trees (Pyrus communis cv. Williams) in orchards (27°46'36.0"N 42°29'44.7"E, 30°00'00.2"N 40°15'11.8"E, and 28°44'52.9"N 36°18'47.8" E) in Hail, Al Jouf, and Tabuk regions exhibited fire blight symptoms. After removing bark, the affected trees showed shoot blight, brown-blighted shoot tips and blossom blight, dead flowers on the stems, and reddish-colored cankers. The disease incidence varied from 10% to 25%. Pathogen was isolated from 21 symptomatic samples including fruits, flowers, and shoots. Bacteria were isolated from washed tissues on King's B (KB) and semi-selective CCT media (Ishimaru and Klos, 1984). After 48 hours, colonies resembling Erwinia amylovora on KB media were 1.5-2 mm in diameter, white, circular, slightly convex, with a smooth surface, and exhibited no fluorescence under ultraviolet light. Colonies on CCT after 72 h were 3-4 mm in diameter, mucoid with shiny surfaces, semitransparent, speckled with craters, and slightly violet. All isolates were purified by subculturing on KB. All isolates were Gram-negative and rod-shaped, fermentative of glucose, positive for catalase and negative for oxidase, and potato rot. They induced a hypersensitive reaction when infiltrated in tobacco leaves (cv. Xanthi). Based on morphology and biochemical tests (EPPO, 2004), the strains were identified as Erwinia sp. Twenty-six strains from Saudi Arabia (SA) and the reference strain (NCPPB 683T) hydrolyzed gelatin and formed white, highly mucous colonies on the levan medium. These strains could not reduce nitrate to nitrite and tested negative for urease and indole production. All the isolates and the reference strain were confirmed to be E. amylovora based on a PCR 0.9-kb DNA fragment amplification with a species-specific primer set, A/B targets pEA29 (Bereswill et al. 1992). 16S-rDNA fragments from Saudi isolates were amplified with 27F and 1492R primers (Lane, 1991). Purified amplicons from PCR were sequenced (OR717505 and OR743536-OR743560), and a BLAST search of the GenBank database revealed 100% (927/927) homology with E. amylovora strain CP066796.1. The housekeeping gene rpoB was PCR amplified with primers CM7-F and CM31b-R (Rezzonico et al. 2012), and the products were sequenced (PP465516-PP465541). BLAST analysis showed 100% (944/944 nt) and 96.19% (908/944 nt) identities with the sequences of E. amylovora ATCC 15580 CP066796.1 and E. pyrifoliae CP201486 CP103445.1, respectively. To fulfill Koch's postulates, SA strains were inoculated on five healthy 3-month-old clones of apple (Malus domestica cv. Gala) per strain with a 10-μl bacterial suspension containing 107 colony-forming units per milliliter by injecting directly in the veins of the upper second leaf plus 5 healthy plants injected with sterile distilled water as control. Plants were incubated at 28°C for six days under a 12-hour light regime. Observed symptoms were similar to the ones observed in the field. The experiment was replicated twice. Bacterial colonies on CCT media were re-isolated from the inoculated apple rootstocks and confirmed by the A/B primer set. To our knowledge, this is the first peer-reviewed report of E. amylovora in SA since the fire blight-like symptoms were observed in SA in 2013 (Alhudaib, 2013). Further research will identify new host plants for the fire blight pathogen within SA which is important due to the importation of pome fruit seedlings (quince, apple, and pear) from neighboring Jordan where E. aylovora was reported (Tehabsim et al. 1992).

N─N Bond Oxidative Addition‐Promoted Diaziridinone Activation: A Mechanistic Study

AbstractDensity functional theory (DFT) calculation has been used to reveal palladium‐catalyzed mode of diaziridinone ring activation. Our theoretical studies found that oxidative addition of di‐tert‐butyldiaziridinone to Pd(II) can give a high valent Pd(IV) intermediate, along with the cleavage of N─N bond in di‐tert‐butyldiaziridinone through a concerted three‐membered‐cyclic transition state. Subsequent transformations via reductive elimination and β‐N elimination give the desired indolo[3,2‐b]indole product. The competitive activation modes such as metathesis and migratory insertion can be excluded in our selected model reaction. The rate‐determining step of this reaction is the oxidative addition of di‐tert‐butyldiaziridinone step. Distortion‐interaction analysis was conducted to reveal that the oxidative addition mode of di‐tert‐butyldiaziridinone is superior to the metathesis mode due to the lower interaction energy.

NHC-Catalyzed Aldimine Umpolung/6π-Electrocyclization Cascade to Access Tetracyclic Dihydrochromeno Indoles.

The umpolung of aldimines using N-heterocyclic carbenes (NHCs) is less explored compared to the established polarity reversal of aldehydes. Described herein is an NHC-catalyzed imine umpolung /6π-electrocyclization cascade, which leads to the atom- and pot-economic synthesis of biologically important dihydrochromeno indoles. For the first time, the nucleophilic aza-Breslow intermediates have been intercepted with unactivated alkynes. Preliminary mechanistic and DFT studies shed light on the role of the phenolic -OH moiety in promoting the addition of the aza-Breslow intermediate to the unactivated alkyne via an intramolecular proton transfer in a stepwise manner. DFT studies also support the regioselectivity preference for the 5-exo-dig cyclization pathway, leading to the exclusive formation of the indole products. Moreover, a comparison of Gibbs free energies provides insight into a thermodynamically preferred 6π-electrocyclization over a competing oxa-Michael pathway. Further, this strategy is applied to the formal synthesis of a Hepatitis C Virus (HCV) NS5A inhibitor in a step-economical method.

NHC‐Catalyzed Aldimine Umpolung/6π‐Electrocyclization Cascade to Access Tetracyclic Dihydrochromeno Indoles

The umpolung of aldimines using N‐heterocyclic carbenes (NHCs) is less explored compared to the established polarity reversal of aldehydes. Described herein is an NHC‐catalyzed imine umpolung /6π‐electrocyclization cascade, which leads to the atom‐ and pot‐economic synthesis of biologically important dihydrochromeno indoles. For the first time, the nucleophilic aza‐Breslow intermediates have been intercepted with unactivated alkynes. Preliminary mechanistic and DFT studies shed light on the role of the phenolic ‐OH moiety in promoting the addition of the aza‐Breslow intermediate to the unactivated alkyne via an intramolecular proton transfer in a stepwise manner. DFT studies also support the regioselectivity preference for the 5‐exo‐dig cyclization pathway, leading to the exclusive formation of the indole products. Moreover, a comparison of Gibbs free energies provides insight into a thermodynamically preferred 6π‐electrocyclization over a competing oxa‐Michael pathway. Further, this strategy is applied to the formal synthesis of a Hepatitis C Virus (HCV) NS5A inhibitor in a step‐economical method.

Enhancing the formation of functional glucosinolate degradation products in fermented broccoli stalk by-product with lactic acid bacteria

Broccoli stalk by-product (BsBP) is rich in glucosinolates (GSLs). Its fermentation process is generally characterized by the degradation of GSLs and formation of bioactive isothiocyanates (ITCs), in which lactic acid bacteria (LAB) play an important role. The GSLs-degrading capacity of 61 LAB strains was investigated in vitro. Lacticaseibacillus paracasei YC5, Pediococcus pentosaceus RBHZ36, and Lactiplantibacillus plantarum ND1, with high potential to transform GSLs into ITCs, were screened. The functional GSL degradation products (total content of sulforaphane, indol-3-carbinol, and ascorbigen) increased 22.0–33.5 % compared to natural fermentation after 24 h when BsBP was fermented by the three screened strains in pure culture. LAB fermentation also helped to increase the quantity of indolic GSL degradation products in BsBP brine, suggesting that LAB fermentation promoted BsBP GSLs transformation into bioactive ITCs. The proposed use of the LAB strains characterized in this study provided a fermented BsBP and brine with high profile of functional GSL degradation products.

First Report of Klebsiella variicola Causing Tomato Pith Necrosis in Yunnan Province, China.

Tomato (Solanum lycopersicum L.) is a key vegetable crop in China. In August 2023, an outbreak of bacterial pith necrosis in tomato occurred in Lufeng County, Yunnan Province, China, affecting over 40% of the tomato plants in a greenhouse. The stems of infected plants developed a waterlogged soft rot and the disease progressed, the lower leaves and lateral branches of infected plants gradually wilted and died. A longitudinal cut of the stem revealed hollow pith with brown vascular tissue. To isolate the pathogen, the plant surface was disinfested with 75% ethanol. Then, a piece of infected tissue from the base of the stem was excised and immersed in sterile water for 2 min. A small amount of liquid was streaked onto TTC (2,3,5-triphenyltetrazolium chloride) agar medium using an inoculation loop, and plates were incubated at 28℃ for 24 h. Colonies on the TTC plate were white, indicating that the pathogen was not Ralstonia solanacearum. Colonies grown on LB (Luria-Bertani) agar medium were randomly selected and subjected to preliminary pathogenicity tests. Based on the results, a colony named Kv4 was selected and purified through six subcultures in LB agar medium. Biochemical tests showed the strain utilized D-sorbitol, raffinose and citrate but not adonitol, and was positive for methyl red, D-glucose (acid), urea hydrolysis, lysine decarboxylase, and motility, and negative for phenylalanine deaminase, H2S production, indole production, and ornithine decarboxylase. These characteristics align with Klebsiella species (Garrity et al. 2007). To determine the species of strain Kv4, partial sequences of the 16S rDNA, phoE, leuS, and rpoB genes were amplified (Barrios-Camacho et al. 2019) and sequenced. Through BLASTn analysis, strain Kv4 sequences of 16S rDNA (OR888750) had 99.47% identity (1488/1496 bp), phoE (OR899599) had 98.69% (605/613 bp) identity, leuS (OR899598) had 99.07% identity (959/968 bp), and rpoB (OR899597) had 97.69% (633/648 bp) identity with Klebsiella variicola strain FF0907. Using the ClustalW algorithm in MEGA11 for nucleotide sequence alignment, phylogenetic trees were constructed with 16S and phoE, leuS, and rpoB via the neighbor-joining method, confirming strain Kv4 as K. variicola. To test pathogenicity, the roots of 25 'Moneymaker' tomato plants with four to five true leaves were wounded, then each plant inoculated with a 15 mL bacterial suspension (OD600=0.6) of strain Kv4, while the control plants received sterile water. Plants were incubated at 28℃ with a 16 h photoperiod. Experiments were done twice. At 15 days after inoculation (DAI), all plants inoculated with Kv4 showed yellowing, unevenly distributed small black necrotic spots on the leaf surface, and purple-brown soft rot at the stem base. By 18 DAI, there was a gradual transformation of the stem bases from green to purplish brown. At 21 DAI, 60% of the inoculated plants displayed brownish soft rot at the stem base. In contrast, the control plants remained symptom-free. The pathogen was re-isolated from the stem and identified as K. variicola via sequence analysis of 16S, phoE, leuS, and rpoB. In recent years, several new bacterial pith necrosis diseases were reported in tomato (Guo et al. 2023; Ivić et al. 2023). This is the first study documenting K. variicola causing bacterial pith necrosis in tomato. Once considered a benign plant endophyte, Sun et al. (2023) reported K. variicola causing banana sheath rot in Guangdong and Guangxi Provinces, China. Malik et al. (2023) reported that K. variicola caused leaf streak in sorghum in India. This report of bacterial pith necrosis in tomato caused by K. variicola strain Kv4 underscores the escalating threat posed by emerging pathogens to agricultural production. The emergence of K. variicola as a tomato pathogen complicates plant disease management strategies.

Regiodivergent Gold-Catalyzed Rearrangement-Addition Reactions of Sulfenylated Propargylic Carboxylates with Indoles.

Sulfenylated propargylic carboxylates were introduced to investigate the influence of sulfur substitution in gold-catalyzed alkyne activation pathways. Regiodivergent gold-catalyzed rearrangement and indole capture reactions proceed under mild conditions to give functionalized indole products bearing sulfenylated (Z)-enol carboxylate motifs. Pathways involving both 1,2- and 1,3-carboxylate migrations are achieved selectively, with indole being added in a 1,4 relationship to the sulfenyl group in each case. High levels of selectivity are influenced by the catalyst system, counterion, and carboxylate group.

Synthesis of Non-canonical Tryptophan Variants via Rh-catalyzed C-H Functionalization of Anilines.

Tryptophan and its non-canonical variants play critical roles in pharmaceutical molecules and enzymes. Facile access to this privileged class of amino acids from readily available building blocks remains a long-standing challenge. Here, we report a regioselective synthesis of non-canonical tryptophans bearing C4-C7 substituents via Rh-catalyzed annulation between structurally diverse tert-butyloxycarbonyl (Boc)-protected anilines and alkynyl chlorides readily prepared from amino acid building blocks. This transformation harnesses Boc-directed C-H metalation and demetalation to afford a wide range of C2-unsubstituted indole products in a redox-neutral fashion. This umpolung approach compared to the classic Larock indole synthesis offers a novel mechanism for heteroarene annulation and will be useful for the synthesis of natural products and drug molecules containing non-canonical tryptophan residues in a highly regioselective manner.

Dynamic changes in volatile profiles and bacterial communities during natural fermentation of Mei yu, traditional Chinese fermented fish pieces

Microbial metabolism is important for the unique flavor formation of Mei yu, a kind of traditional Chinese fermented fish pieces. However, the interactive relationship between microorganisms and flavor components during fermentation is still unclear. In this study, electronic nose and headspace-solid-phase microextraction-gas chromatography-mass spectrometry analysis were performed to identify flavor components in Mei yu during the fermentation, and the absolute microbial quantification was conducted to identify the diversity and succession of microbial communities. During fermentation, there was an increase in the types of volatile compounds. Alcohols, aldehydes, aromatics and esters were the main flavor compounds and significantly increased in Mei yu, while hydrocarbon and aldehydes significantly decreased. The absolute abundances of Lactobacillus, Lactococcus and Weissella increased significantly after 3 days’ fermentation, which were closely associated with the productions of 1-nonanol, 2-methoxy-4-vinylphenol, guaiacol, ethyl palmitate and ethyl caprylate that might though pathways related to fatty acid biosynthesis and amino acid metabolism. However, these genera were negatively correlated with the production of indole. Additionally, the total volatile basic nitrogen (TVB-N) levels of Mei yu fermented during 3 days were within the limits of 25 mg TVB-N/100 g fish, with the contents of free amino acids and lipoxygenase activities were significant lower than that of 4 days’ fermentation. In view of food safety and flavor, it suggested that the natural fermented Mei yu at room temperature should be controlled within 3 days. This study highlights the application of absolute quantification to microbiome analysis in traditional fermented Mei yu and provides new insights into the roles of microorganisms in flavor formation during fermentation.

Evaluation and Characterization of Rhizobacteria and Endophytes for Antagonistic and Growth Promotional Traits Against Collar Rot (Sclerotium rolfsii Sacc.) of Soybean

Background: Sclerotium rolfsii Sacc. is one of the destructive soil-borne plant pathogens due to its significant loss in terms of yield and quality in several crop plants including soybean. Use of biocontrol agents in the management of disease is an ecofriendly strategy. Therefore, the study aimed to assess in vitro antagonisticactivity of rhizobacteria and endophytes against Sclerotium rolfsii following dual culture technique. Methods: A total of 75 rhizobacterial isolates, 3 endophytes along with reference cultures were evaluated for the antagonistic activity against Sclerotium rolfsii following dual culture technique. The effective rhizobacterial isolates were tested for the various plat growth promoting rhizobacteria traits viz., indole acetic acid production, siderophore production, Phosphate solubilization, production of ammonia, hydrogen cyanide production and chitinase test following the standard procedure. Result: The two promising rhizobacterial strains i.e., AUUB 209 (Streptomyces enissocaesilis) and Streptomyces racemochromogenes (AUDT 626) were found effective with 76.79 and 64.75 percent mycelial inhibition of Sclerotium rolfsii under in vitro. The antagonistic activity of these two rhizobacteria is mainly attributed to the positive confirmation of various PGPR activities viz., production of Indole acetic acid, siderophore, phosphate solubilization and chitinase production.

Termite gut microbiome being a rich source of potential lignocellulolytic bacterial strains

A study was carried out to identify prospective cellulose degrading bacteria isolated from termite gut collected from various agriculture sites. Eight of the bacterial isolates displayed a very distinct zone of clearing on CMC (Carboxylmethyl cellulose) agar media after staining with 1% Congo red, indicating that they may have cellulose degrading ability. On a CMC agar plate, CRDB6 had the largest colony diameter (3.08 cm) as well as clear zone diameter (0.96 cm), however CRDB 1 has maximum cellulolytic activity (1.45 cm) followed by CRDB5 and CRDB6 (1.33 cm and 1.28 cm) respectively. CRDB3, CRDB4 and CRDB9 showed Gram negative reaction among crop residue degrading bacterial isolates, while the remainder showed Gram positive reaction. The morphological characteristics of the bacteria isolates, such as colony shape, elevation, margin, colour and opacity, differed. Isolates CRDB6 produced highest β-glucosidase activity (12.66 m L-1) followed by CRDB1 (12.20 mg m L-1) and CRDB5 (10.21 m L-1) in Luria broth culture. Indole acetic acid production (with tryptophan) was greater in CRDB1 (389.33 mg/ml), CRDB6 (356.9 m L-1) and CRDB3 (315.96 m L-1) while as P-solubilization potential was peaked in CRDB6 (2.41 m L-1) followed by CRDB4 (1.76 m L-1) and CRDB3. Overall, isolate CRDB6 exhibited the highest cellulolytic activity as compared to rest of the bacterial isolates. These results will help in preparing bacterial consortia for the management of crop residues.

Chengkuizengella axinellae sp. nov., a symbiotic bacterium isolated from a marine sponge of the genus Axinella.

A Gram-stain-positive, strictly aerobic, creamy-white colored, endospore-forming and non-motile rods strain, designated as strain 2205SS18-9T, was isolated from a marine sponge, Axinella sp. collected from Seopseom Island, Republic of Korea. Optimal growth of strain 2205SS18-9T was observed at 25-30°C, pH 6.5-7.0, and in the presence of 3.0% (w/v) NaCl. Cells were oxidase-positive and catalase-negative. Negative for nitrate reduction and indole production. Phylogenetic analyses based on the 16S rRNA gene and whole-genome sequences revealed that strain 2205SS18-9T formed a distinct phyletic lineage in the genus Chengkuizengella, and it was most closely related to Chengkuizengella marina YPA3-1-1T and Chengkuizengella sediminis J15A17T with 97.1 and 96.6% 16S rRNA gene sequence similarities, respectively. The average nucleotide identity and digital DNA-DNA hybridization values between strain 2205SS18-9T and Chengkuizengella marina YPA3-1-1T were 79.0 and 21.6%, respectively. The genomic DNA G + C content was 34.1%. The genome harbors a number of host-adhesion and transporter genes, suggested that strain 2205SS18-9T may interact with its sponge host as a symbiont. Menaquinone-7 was the sole isoprenoid quinone and antieiso-C15:0 (28.5%), iso-C16:0 (25.8%), C16:1 ω7c alcohol (15.0%), and iso-C15:0 (11.2%) were detected as the major fatty acids. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified aminophospholipids, and an unidentified lipid. The cell-wall peptidoglycan contained lysine, alanine, glutamate, and aspartate. Based on these analyses, strain 2205SS18-9T represents a novel species of the genus Chengkuizengella, for which the name Chengkuizengella axinellae sp. nov. is proposed. The type strain is 2205SS18-9T (= KACC 23238T = LMG 33063T).

Plant-Growth-Promoting Rhizobacteria Improve Seeds Germination and Growth of Argania spinosa.

Argania spinosa is among the most important species of the Moroccan forest in terms of ecological, environmental, and socio-economic aspects. However, it faces a delicate balance between regeneration and degradation in its natural habitat. Hence, the efforts to preserve and regenerate argan forests are crucial for biodiversity, soil quality, and local livelihoods, yet they face challenges like overgrazing and climate change. Sustainable management practices, including reforestation and community engagement, are vital for mitigating degradation. Similarly, exploiting the argan tree's rhizosphere can enhance soil quality by leveraging its rich microbial diversity. This approach not only improves crop growth but also maintains ecosystem balance, ultimately benefiting both agriculture and the environment. This enrichment can be achieved by different factors: mycorrhizae, plant extracts, algae extracts, and plant growth-promoting rhizobacteria (PGPR). The benefits provided by PGPR may include increased nutrient availability, phytohormone production, shoot, root development, protection against several plant pathogens, and disease reduction. In this study, the effect of rhizobacteria isolated from the Agran rhizosphere was evaluated on germination percentage and radicle length for Argania spinosa in vitro tests, growth, collar diameter, and branching number under greenhouse conditions. One hundred and twenty (120) bacteria were isolated from the argan rhizosphere and evaluated for their capacity for phosphate solubilization and indole acetic acid production. The results showed that 52 isolates could solubilize phosphorus, with the diameters of the solubilization halos varying from 0.56 ± 0.14 to 2.9 ± 0.08 cm. Among 52 isolates, 25 were found to be positive for indole acetic acid production. These 25 isolates were first tested on maize growth to select the most performant ones. The results showed that 14 isolates from 25 tested stimulated maize growth significantly, and 3 of them by 28% (CN005, CN006, and CN009) compared to the control. Eight isolates (CN005, CN006, CN004, CN007, CN008, CN009, CN010, and CN011) that showed plant growth of more than 19% were selected to evaluate their effect on argan germination rate and radicle length and were subjected to DNA extraction and conventional Sanger sequencing. The 8 selected isolates were identified as: Brevundimonas naejangsanensis sp2, Alcaligenes faecalis, Brevundimonas naejangsanensis sp3, Brevundimonas naejangsanensis sp4, Leucobacter aridicollis sp1, Leucobacter aridicollis sp2, Brevundimonas naejangsanensis sp1, and Staphylococcus saprophyticus. The results showed that Leucobacter aridicollis sp2 significantly increased the germination rate by 95.83%, and the radicle length with a value of 2.71 cm compared to the control (1.60 cm), followed by Brevundimonas naejangsanensis sp3 and Leucobacter aridicollis sp1 (2.42 cm and 2.11 cm, respectively). Under greenhouse conditions, the results showed that the height growth increased significantly for Leucobacter aridicollis sp1 (42.07%) and Leucobacter aridicollis sp2 (39.99%). The isolates Brevundimonas naejangsanensis sp3 and Leucobacter aridicollis sp1 increased the gain of collar diameter by 41.56 and 41.21%, respectively, followed by Leucobacter aridicollis sp2 and Staphyloccocus saprophyticus (38.68 and 22.79%). Leucobacter aridicollis sp1 increased the ramification number per plant to 12 compared to the control, which had 6 ramifications per plant. The use of these isolates represents a viable alternative in sustainable agriculture by improving the germination rate and root development of the argan tree, as well as its development, while increasing the availability of nutrients in the soil and consequently improving fertilization.

A novel exopolysaccharide-producing bacterium, Pseudescherichia liriopis sp. nov. isolated from Liriope platyphylla, enhances the growth of Daucus carota subsp. sativus under drought and salinity stress.

Biological and abiotic stresses in plant growth are associated with reduced crop yields. Therefore, improving plant stress resistance can be a crucial strategy to improve crop production. To overcome these problems, plant growth-promoting bacteria are emphasized as one of the alternative tools for sustainable agriculture. This study found a novel strain (L3T) of a plant growth-promoting bacterium in fermented Liriope platyphylla fruit. Strain L3T showed the ability to promote plant growth. The L3T strain promoted plant growth of D. carota subsp. sativus, increasing the length (increase rate compared to the control group, 36.98%), diameter (47.06%), and weight of carrots (81.5%), ultimately increasing the edible area. In addition, we confirmed that plant growth was improved even in situations that inhibited plant growth, such as salinity and drought stress. Strain L3T performed indole production, siderophore production, phosphate solubilization, and nitrogen fixation, all characteristics of a strain that promotes plant growth. Genome analysis revealed genes involved in the growth promotion effects of strain L3T. Additionally, the properties of exopolysaccharides were identified and characterized using FTIR, TGA, and UHPLC. Our results demonstrated that L3 isolated from fermented L. platyphylla fruit can be used to simultaneously alleviate drought and NaCl stress.

Optimisation of indole acetic acid production by Neopestalotiopsis aotearoa endophyte isolated from Thymus vulgaris and its impact on seed germination of Ocimum basilicum

BackgroundMicrobial growth during plant tissue culture is a common problem that causes significant losses in the plant micro-propagation system. Most of these endophytic microbes have the ability to propagate through horizontal and vertical transmission. On the one hand, these microbes provide a rich source of several beneficial metabolites.ResultsThe present study reports on the isolation of fungal species from different in vitro medicinal plants (i.e., Breynia disticha major, Breynia disticha, Duranta plumieri, Thymus vulgaris, Salvia officinalis, Rosmarinus officinalis, and Ocimum basilicum l) cultures. These species were tested for their indole acetic acid (IAA) production capability. The most effective species for IAA production was that isolated from Thymus vulgaris plant (11.16 µg/mL) followed by that isolated from sweet basil plant (8.78 µg/mL). On screening for maximum IAA productivity, medium, “MOS + tryptophan” was chosen that gave 18.02 μg/mL. The macroscopic, microscopic examination and the 18S rRNA sequence analysis indicated that the isolate that given code T4 was identified as Neopestalotiopsis aotearoa (T4). The production of IAA by N. aotearoa was statistically modeled using the Box-Behnken design and optimized for maximum level, reaching 63.13 µg/mL. Also, IAA extract was administered to sweet basil seeds in vitro to determine its effect on plant growth traits. All concentrations of IAA extract boosted germination parameters as compared to controls, and 100 ppm of IAA extract exhibited a significant growth promotion effect for all seed germination measurements.ConclusionsThe IAA produced from N. aotearoa (T4) demonstrated an essential role in the enhancement of sweet basil (Ocimum basilicum) growth, suggesting that it can be employed to promote the plant development while lowering the deleterious effect of using synthetic compounds in the environment.

Neisseria leonii sp. nov., isolated from the nose, lung, and liver of rabbits.

A taxogenomic study of three strains (3986T, 51.81, and JF 2415) isolated from rabbits between 1972 and 2000 led to the description of a new Neisseria species. The highest sequence similarity of the 16S rRNA gene was found to Neisseria animalis NCTC 10212T (96.7 %). The 16S rRNA gene similarity above 99 % and average nucleotide identity (ANI) values above 96 % among the strains, indicated that they belong to the same species. At the same time, the strains shared ANI values below 81 % and dDDH values below 24 % with all described Neisseria species. In the bac120 gene phylogenetic tree, the three strains clustered near Neisseria elongata and Neisseria bacilliformis in the Neisseria clade. However, the Neisseria clade is not monophyletic, and includes the type strains of Morococcus cerebrosus, Bergeriella denitrificans, Kingella potus, Uruburuella suis, and Uruburuella testudinis. Neisseria shayeganii clustered outside the clade with members of the genus Eikenella. Amino acid identity (AAI) values were calculated, and a threshold of 71 % was used to circumscribe the genus Neisseria. According to this proposed AAI threshold, strains 3986T, 51.81, and JF 2415 were placed within the genus Neisseria. The cells of the three strains were Gram-stain-negative diplococcobacilli and non-motile. Optimal growth on trypticase soy agar occurred at 37 °C and pH 8.5 in aerobic conditions. Notably, all strains exhibited indole production in the API-NH test, which is atypical for Neisseria and the family Neisseriaceae. The strains exhibited a common set of 68 peaks in their MALDI-TOF MS profiles, facilitating the swift and accurate identification of this species. Based on genotypic and phenotypic data, it is proposed that strains 3986T, 51.81, and JF 2415 represent a novel species within the genus Neisseria, for which the name Neisseria leonii sp. nov. is proposed (type strain 3986T=R726T=CIP 109994T=LMG 32907T).

Dietary fibre directs microbial tryptophan metabolism via metabolic interactions in the gut microbiota

Tryptophan is catabolized by gut microorganisms resulting in a wide range of metabolites implicated in both beneficial and adverse host effects. How gut microbial tryptophan metabolism is directed towards indole, associated with chronic kidney disease, or towards protective indolelactic acid (ILA) and indolepropionic acid (IPA) is unclear. Here we used in vitro culturing and animal experiments to assess gut microbial competition for tryptophan and the resulting metabolites in a controlled three-species defined community and in complex undefined human faecal communities. The generation of specific tryptophan-derived metabolites was not predominantly determined by the abundance of tryptophan-metabolizing bacteria, but rather by substrate-dependent regulation of specific metabolic pathways. Indole-producing Escherichia coli and ILA- and IPA-producing Clostridium sporogenes competed for tryptophan within the three-species community in vitro and in vivo. Importantly, fibre-degrading Bacteroides thetaiotaomicron affected this competition by cross-feeding monosaccharides to E. coli. This inhibited indole production through catabolite repression, thus making more tryptophan available to C. sporogenes, resulting in increased ILA and IPA production. The fibre-dependent reduction in indole was confirmed using human faecal cultures and faecal-microbiota-transplanted gnotobiotic mice. Our findings explain why consumption of fermentable fibres suppresses indole production but promotes the generation of other tryptophan metabolites associated with health benefits.

Photorhabdus africana sp. nov. isolated from Heterorhabditis entomopathogenic nematodes

One Gram-negative, rod-shaped bacterial strain, isolated from an undescribed Heterorhabditis entomopathogenic nematode species was characterized to determine its taxonomic position. The 16S rRNA gene sequences indicate that it belongs to the class Gammaproteobacteria, to the family Morganellaceae, to the genus Photorhabdus, and likely represents a novel bacterial species. This strain, designated here as CRI-LCT, was therefore molecularly, biochemically, and morphologically characterized to describe the novel bacterial species. Phylogenetic reconstructions using 16S rRNA gene sequences show that CRI-LCT is closely related to P. laumondii subsp. laumondii TT01T and to P. laumondii subsp. clarkei BOJ-47T. The 16rRNA gene sequences between CRI-LCT and P. laumondii subsp. laumondii TT01T are 99.1% identical, and between CRI-LCT and P. laumondii subsp. clarkei BOJ-47T are 99.2% identical. Phylogenetic reconstructions using whole genome sequences show that CRI-LCT is closely related to P. laumondii subsp. laumondii TT01T and to P. laumondii subsp. clarkei BOJ-47T. Moreover, digital DNA-DNA hybridization (dDDH) values between CRI-LCT and its two relative species P. laumondii subsp. laumondii TT01T and P. laumondii subsp. clarkei BOJ-47T are 65% and 63%, respectively. In addition, we observed that average nucleotide identity (ANI) values between CRI-LCT and its two relative species P. laumondii subsp. laumondii TT01T and P. laumondii subsp. clarkei BOJ-47T are 95.8% and 95.5%, respectively. These values are below the 70% dDDH and the 95–96% ANI divergence thresholds that delimits prokaryotic species. Based on these genomic divergence values, and the phylogenomic separation, we conclude that CRI-LCT represents a novel bacterial species, for which we propose the name Photorhabdus africana sp. nov. with CRI-LCT (= CCM 9390T = CCOS 2112T) as the type strain. The following biochemical tests allow to differentiate P. africana sp. nov. CRI-LCT from other species of the genus, including its more closely related taxa: β-Galactosidase, citrate utilization, urease and tryptophan deaminase activities, indole and acetoin production, and glucose and inositol oxidation. Our study contributes to a better understanding of the taxonomy and biodiversity of this important bacterial group with great biotechnological and agricultural potential.

Palladium(0)‐catalyzed Synthesis of Indoles and Carbazoles from o‐ Alkenyl (Alkyl) Anilines via C–H Amination Cyclization

Various indoles and carbazoles were synthesized from o‐alkenyl anilines and o‐aryl anilines by the β‐C(sp2)‐H bond amination cyclization without any oxidants or hydrogen acceptors. Likewise, employing o‐alkyl anilines as substrates, indole products are also obtained by the β‐C(sp3)‐H bond amination cyclization, and dehydrogenation. In this cross‐dehydrogenative coupling, many different functional groups are tolerated, and hydrogen gas is the only by‐product, thereby providing an intrinsically waste‐free approach for the synthesis of indoles and carbazoles.

Exploring Endophytic Bacteria from Artemisia spp. and Beneficial Traits on Pea Plants.

Endophytic microorganisms represent promising solutions to environmental challenges inherent in conventional agricultural practices. This study concentrates on the identification of endophytic bacteria isolated from the root, stem, and leaf tissues of four Artemisia plant species. Sixty-one strains were isolated and sequenced by 16S rDNA. Sequencing revealed diverse genera among the isolated bacteria from different Artemisia species, including Bacillus, Pseudomonas, Enterobacter, and Lysinibacillus. AR11 and VR24 obtained from the roots of A. absinthium and A. vulgaris demonstrated significant inhibition on Fusarium c.f. oxysporum mycelial growth. In addition, AR11, AR32, and CR25 exhibited significant activity in phosphatase solubilization, nitrogen fixation, and indole production, highlighting their potential to facilitate plant growth. A comparative analysis of Artemisia species showed that root isolates from A. absinthium, A. campestris, and A. vulgaris have beneficial properties for inhibiting pathogen growth and enhancing plant growth. AR11 with 100% similarity to Bacillus thuringiensis, could be considered a promising candidate for further investigation as microbial biofertilizers. This finding highlights their potential as environmentally friendly alternatives to chemical pesticides, thereby contributing to sustainable crop protection practices.