Endophytic Actinomycetes Research Articles

Streptomyces albidocamelliae sp. nov., an endophytic actinomycete isolated from the leaves of Camellia oleifera.

A novel actinobacterium strain, HUAS 14-6T, was isolated from the healthy leaves of Camellia oleifera collected from Changde City, Hunan Province, China. Strain HUAS 14-6T produced tight spiral spore chains consisting of oval or spherical spores with a smooth surface. 16S rRNA gene sequence analysis revealed that strain HUAS 14-6T belonged to the genus Streptomyces and shared highest similarity to Streptomyces bungoensis DSM 41781T (99.72%). Phylogenetic analysis based on 16S rRNA gene sequences indicated strain HUAS 14-6T was in a clade with S. bungoensis DSM 41781T. However, the ANIm and dDDH between strain HUAS 14-6T and S. bungoensis DSM 41781T were 88.16% and 31.2%, respectively, far less than the species-level thresholds. Phylogenetic trees based on the five housekeeping genes (atpD, gyrB, recA, rpoB and trpB) showed that strain HUAS 14-6T formed a separate branch, indicating that this strain could belong to a potential new species. Pairwise MLSA distances between strain HUAS 14-6T and all type strains exhibiting 16S rRNA gene sequence similarities of ≥98.7% to it were much higher than the maximum range of 0.014 recommended for delineating a new Streptomyces species. Based on polyphasic taxonomic study, HUAS 14-6T represents a novel species within the genus Streptomyces for which the name Streptomyces albidocamelliae sp. nov. is proposed. The type strain is HUAS 14-6T (=MCCC 1K08365T=JCM 35920T).

Endophytic actinomycetes promote growth and fruits quality of tomato (Solanum lycopersicum): an approach for sustainable tomato production.

Tomato, a fruit with a high vitamin content, is popular for consumption and economically important in Thailand. However, in the past year, the extensive usage of chemicals has significantly decreased tomato yields. Plant Growth-Promoting Rhizobacteria (PGPR) is an alternative that can help improve tomato production system growth and yield quality while using fewer chemicals. The present study aimed to determine whether endophytic actinomycetes promote growth and fruit quality of tomato (Solanum lycopersicum). The experiment was conducted in a net-houses at the Center for Agricultural Resource System Research, Faculty of Agriculture, Chiang Mai University, Chiang Mai province, Thailand. The randomized completely block design (RCBD) was carried out for four treatments with three replications, which was control, inoculation with TGsR-03-04, TGsL-02-05 and TGsR-03-04 with TGsL-02-05 in tomato plant. Isolated Actinomycetes spp. of each treatment was then inoculated into the root zone of tomato seedlings and analyzed by Scanning Electron Microscopy (SEM). The height of tomato plants was measured at 14, 28, 56, and 112 days after transplanting. Final yield and yield quality of tomato was assessed at the maturity phase. The SEM result illustrated that the roots of tomato seedlings from all treatments were colonized by endophytic actinomycetes species. This contributed to a significant increase in plant height at 14 days after transplanting (DAT), as found in the TGsR-03-04 treatment (19.40 cm) compared to the control. Besides, all inoculated treatments enhanced tomato yield and yield quality. The highest weight per fruit (47.38 g), fruit length (52.37 mm), vitamin C content (23.30 mg 100g-1), and lycopene content (145.92µg g-1) were obtained by inoculation with TGsR-03-04. Moreover, the highest yield (1.47 kg plant-1) was obtained by inoculation with TGsL-02-05. There was no statistically significant difference in the number of fruits per plant, fruit width, brix, and antioxidant activity when various inoculations of endophytic actinomycetes were applied. Therefore, the use of endophytic actinomycetes in tomato cultivation may be an alternative to increase tomato yield and yield quality.

Identification of Endophytic Actinomycetes Isolated From Hopea ferrea and Its Antibacterial Activity Against Cariogenic Bacterium

Dental caries affects around 36% of the world's population and results in the loss of primary teeth in about 530 million youngsters. It is described as the loss of the enamel layer of the tooth by acids generated by the activity of cariogenic bacterium such as Streptococcus mutans on carbohydrates. This study aims to identify bioactive endophytic actinomycetes as an antibacterial agent against S. mutans DSM 20523 and to identify the bioactive endophytic actinomycetes using the 16S rRNA sequencing approach. To determine the percentage zone of inhibition, three endophytes at 10-day-old culture have been streaked onto ISP2. Then, S. mutans was streaked perpendicular to the endophytes. Crude extracts from three endophytes have been prepared by using four different types of media namely Tryptic Soy broth, Kings B, International Streptomyces Project Medium No.1 (ISP1) and Starch Casein broth. These crude extracts proceeded to undergo disk diffusion and MIC assays against S. mutans. All three endophytes from streaked plates with the highest percentage zone of inhibition were identified using the 16S rRNA gene molecular approach for characterisation. Endophytic actinomycetes which were isolated from Hopea ferrea showed potent antibacterial activity against S. mutans. All endophytic actinomycetes isolates from streak plates showed good inhibitory activity against the S. mutans. The highest percentage of inhibition was shown by the PT9-13S2 isolate followed by PT9-8S2 and PT9-13W2. Furthermore, crude extracts of PT9-8S2 and PT9-13W2 (the highest ZOI) from TSB media showed inhibitory activities against S. mutans, from disk diffusion as well as minimum inhibitory assays. From the 16S rRNA molecular approach, endophytic actinomycetes PT9-8S2 was closely related to Streptomyces collinus while PT9-13S2 and PT9-13W2 were closely related to Streptomyces malachitospinus. From this study, endophytic actinomycetes showed a promising source as antibacterial agents against cariogenic bacterium S. mutans which is the main causative agent of dental caries.

Antibiotic production by an endophytic Streptomyces isolated from the medicinal plant Poikilospermum suaveolens

Abstract. Ali A, Passitta M, Rante H, Wahyudin E, Djide NJN, Politan RJ, Nur EA, Shigeno S, Ohte S, Kobayashi K, Hosoda K, Tomoda H, Ohshiro T. 2024. Antibiotic production by an endophytic Streptomyces isolated from the medicinal plant Poikilospermum suaveolens. Biodiversitas 25: 2221-2229. This study investigates the antibiotic production potential of endophytic actinomycetes isolated from Poikilospermum suaveolens plants. Samples were collected from various parts of the plant in karst ecosystems. Endophytic actinomycetes were cultured and screened for antimicrobial activity using agar diffusion and liquid microdilution methods. One of the 11 isolated strains exhibited significant antibiotic activity against clinically relevant organisms identified as belonging to the Streptomyces genus, specifically Streptomyces mirabilis strain NRRL ISP-5169. Submerged fermentation of this strain over 9 days showed strong antagonistic effects against Gram-positive bacteria. Spectroscopic analyses confirmed the presence of actinomycin D as the bioactive compound. This study provides on the previously unexplored potential of endophytic actinomycetes from P. suaveolens as a promising resource for antibiotic discovery.

Kitasatospora cathayae sp. nov., a novel endophytic actinomycete isolated from the leaves of Cathaya argyrophylla.

Strain HUAS 3-15T was isolated from the leaves of Cathaya argyrophylla collected from Chenzhou, Hunan Province, PR China. The main fatty acids (>5.0 %) of the strain were anteiso-C15 : 0, C16 : 0, C18 : 1 ω9c, iso-C16 : 0, summed feature 5 (C18 : 2 ω6,9c/C18 : 0 ante), iso-C15 : 0 and anteiso-C17 : 0. MK-9(H6), MK-9(H8) and MK-9(H4) were detected as respiratory quinones. The diagnostic cell-wall diamino acid was meso-diaminopimelic acid. Galactose, glucose and ribose were also present in the cell wall. The major polar lipids consisted of diphosphatidylglycerol, phosphatidyl ethanolamine, phosphatidylinositol mannosides and unidentified phospholipids. The DNA G+C content of the genome sequence, consisting of 8 860 963 bp, is 72.4 mol%. blast analysis based on 16S rRNA gene sequences revealed that the strain belongs to the genus Kitasatospora, with 99.37, 99.03, 98.95, 98.68 and 98.67 % sequence similarity to Kitasatospora aureofaciens ATCC 10762T, Kitasatospora viridis DSM 44826T, Kitasatospora xanthocidica NBRC 13469T, Kitasatospora aburaviensis NRRL B-2218T and Kitasatospora kifunensis IFO 15206T, respectively. Phylogenetic trees based on 16S rRNA gene and whole-genome sequences demonstrated that strain HUAS 3-15T formed a well-supported cluster with K. aureofaciens ATCC 10762T. Further genomic characterization through average nucleotide identity (ANIb/m) and digital DNA-DNA hybridization analysis between strain HUAS 3-15T and K. aureofaciens ATCC 10762T showed values of 90.62/92.55 % and 45.3 %, respectively, lower than the 95-96 % ANI threshold and 70.0 % cutoff used as guideline values for species delineation in bacteria. Furthermore, the differences between the strain and its phylogenomic neighbour in terms of physiological (e.g. sole carbon source growth) and chemotaxonomic (e.g. cellular fatty composition) characteristics further supported this conclusion. Consequently, we concluded that strain HUAS 3-15T represents a novel species of the genus Kitasatospora, for which the name Kitasatospora cathayae sp. nov. is proposed. The type strain is HUAS 3-15T (=MCCC 1K08542T=JCM 36274T).

Actinomycetes are a natural resource for sustainable pest control and safeguarding agriculture.

Actinomycetes, a diverse group of bacteria with filamentous growth characteristics, have long captivated researchers and biochemists for their prolific production of secondary metabolites. Among the myriad roles played by actinomycete secondary metabolites, their historical significance in the field of biocontrol stands out prominently. The fascinating journey begins with the discovery of antibiotics, where renowned compounds like streptomycin, tetracycline, and erythromycin revolutionized medicine and agriculture. The history of biocontrol traces its roots back to the early twentieth century, when scientists recognized the potential of naturally occurring agents to combat pests and diseases. The emergence of synthetic pesticides in the mid-twentieth century temporarily overshadowed interest in biocontrol. However, with growing environmental concerns and the realization of the negative ecological impacts of chemical pesticides, the pendulum swung back towards exploring sustainable alternatives. Beyond their historical role as antibiotics, actinomycete-produced secondary metabolites encompass a rich repertoire with biopesticide potential. The classification of these compounds based on chemical structure and mode of action is highlighted, demonstrating their versatility against both plant pathogens and insect pests. Additionally, this review provides in-depth insights into how endophytic actinomycete strains play a pivotal role in biocontrol strategies. Case studies elucidate their effectiveness in inhibiting Spodoptera spp. and nematodes through the production of bioactive compounds. By unraveling the multifunctional roles of endophytic actinomycetes, this review contributes compelling narrative knowledge to the field of sustainable agriculture, emphasizing the potential of these microbial allies in crafting effective, environmentally friendly biocontrol strategies for combating agricultural pests.

Multifunctional in vitro, in silico and DFT analyses on antimicrobial BagremycinA biosynthesized by Micromonospora chokoriensis CR3 from Hieracium canadense

Among the actinomycetes in the rare genera, Micromonospora is of great interest since it has been shown to produce novel therapeutic compounds. Particular emphasis is now on its isolation from plants since its population from soil has been extensively explored. The strain CR3 was isolated as an endophyte from the roots of Hieracium canadense, and it was identified as Micromonospora chokoriensis through 16S gene sequencing and phylogenetic analysis. The in-vitro analysis of its extract revealed it to be active against the clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Candida tropicalis (15 mm). No bioactivity was observed against Gram-negative bacteria, Escherichia coli ATCC 25922, and Klebsiella pneumoniae ATCC 706003. The Micromonospora chokoriensis CR3 extract was also analyzed through the HPLC-DAD-UV–VIS resident database, and it gave a maximum match factor of 997.334 with the specialized metabolite BagremycinA (BagA). The in-silico analysis indicated that BagA strongly interacted with the active site residues of the sterol 14-α demethylase and thymidylate kinase enzymes, with the lowest binding energies of − 9.7 and − 8.3 kcal/mol, respectively. Furthermore, the normal mode analysis indicated that the interaction between these proteins and BagA was stable. The DFT quantum chemical properties depicted BagA to be reasonably reactive with a HOMO-LUMO gap of (ΔE) of 4.390 eV. BagA also passed the drug-likeness test with a synthetic accessibility score of 2.06, whereas Protox-II classified it as a class V toxicity compound with high LD50 of 2644 mg/kg. The current study reports an endophytic actinomycete, M. chokoriensis, associated with H. canadense producing the bioactive metabolite BagA with promising antimicrobial activity, which can be further modified and developed into a safe antimicrobial drug.

Comparative genomics reveals insight into the phylogeny and habitat adaptation of novel Amycolatopsis species, an endophytic actinomycete associated with scab lesions on potato tubers.

A novel endophytic actinomycete, strain MEP2-6T, was isolated from scab tissues of potato tubers collected from Mae Fag Mai Sub-district, San Sai District, Chiang Mai Province, Thailand. Strain MEP2-6T is a gram-positive filamentous bacteria characterized by meso-diaminopimelic acid in cell wall peptidoglycan and arabinose, galactose, glucose, and ribose in whole-cell hydrolysates. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and hydroxy-phosphatidylethanolamine were the major phospholipids, of which MK-9(H6) was the predominant menaquinone, whereas iso-C16:0 and iso-C15:0 were the major cellular fatty acids. The genome of the strain was 10,277,369 bp in size with a G + C content of 71.7%. The 16S rRNA gene phylogenetic and core phylogenomic analyses revealed that strain MEP2-6T was closely related to Amycolatopsis lexingtonensis NRRL B-24131T (99.4%), A. pretoriensis DSM 44654T (99.3%), and A. eburnea GLM-1T (98.9%). Notably, strain MEP2-6T displayed 91.7%, 91.8%, and 87% ANIb and 49%, 48.8%, and 35.4% dDDH to A. lexingtonensis DSM 44653T (=NRRL B-24131T), A. eburnea GLM-1T, and A. pretoriensis DSM 44654T, respectively. Based on phenotypic, chemotaxonomic, and genomic data, strain MEP2-6T could be officially assigned to a novel species within the genus Amycolatopsis, for which the name Amycolatopsis solani sp. nov. has been proposed. The type of strain is MEP2-6T (=JCM 36309T = TBRC 17632T = NBRC 116395T). Amycolatopsis solani MEP2-6T was strongly proven to be a non-phytopathogen of potato scab disease because stunting of seedlings and necrotic lesions on potato tuber slices were not observed, and there were no core biosynthetic genes associated with the BGCs of phytotoxin-inducing scab lesions. Furthermore, comparative genomics can provide a better understanding of the genetic mechanisms that enable A. solani MEP2-6T to adapt to the plant endosphere. Importantly, the strain smBGCs accommodated 33 smBGCs encoded for several bioactive compounds, which could be beneficially applied in the fields of agriculture and medicine. Consequently, strain MEP2-6T is a promising candidate as a novel biocontrol agent and antibiotic producer.

Antimicrobial Action Mechanisms of Natural Compounds Isolated from Endophytic Microorganisms.

Infectious diseases are a significant challenge to global healthcare, especially in the face of increasing antibiotic resistance. This urgent issue requires the continuous exploration and development of new antimicrobial drugs. In this regard, the secondary metabolites derived from endophytic microorganisms stand out as promising sources for finding antimicrobials. Endophytic microorganisms, residing within the internal tissues of plants, have demonstrated the capacity to produce diverse bioactive compounds with substantial pharmacological potential. Therefore, numerous new antimicrobial compounds have been isolated from endophytes, particularly from endophytic fungi and actinomycetes. However, only a limited number of these compounds have been subjected to comprehensive studies regarding their mechanisms of action against bacterial cells. Furthermore, the investigation of their effects on antibiotic-resistant bacteria and the identification of biosynthetic gene clusters responsible for synthesizing these secondary metabolites have been conducted for only a subset of these promising compounds. Through a comprehensive analysis of current research findings, this review describes the mechanisms of action of antimicrobial drugs and secondary metabolites isolated from endophytes, antibacterial activities of the natural compounds derived from endophytes against antibiotic-resistant bacteria, and biosynthetic gene clusters of endophytic fungi responsible for the synthesis of bioactive secondary metabolites.

Myceligenerans pegani sp. nov., an endophytic actinomycete isolated from Peganum harmala L. in Xinjiang, PR China.

An endophytic actinomycete designated TRM65318T, was isolated from the root of Peganum harmala L. Its taxonomic status was determined using a polyphasic approach. Comparative 16S rRNA gene sequence analysis indicated that strain TRM65318T is phylogenetically most closely related to Myceligenerans salitolerans XHU 5031T (98.15 %) and Myceligenerans xiligouense DSM 15700T (97.78 %). The peptidoglycan belonged to type A4α. The polar lipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, two unknown lipids and three glycolipids. The predominant menaquinones were MK-9(H4) and MK-9(H6) and the whole-cell sugars contained glucose, mannose and galactose. Major fatty acids were anteiso-C15 : 0, iso-C15 : 0 and C16 : 0. Strain TRM65318T had a genome size of 5881012 bp with a genome G+C content of 71.79 mol%. The average nucleotide identity and DNA-DNA hybridization values between strain TRM65318T and the most closely related species were much lower than the thresholds commonly used to define species. At the same time, differences in phenotypic and genotypic data showed that strain TRM65318T could be clearly distinguished from M. salitolerans XHU 5031T. Therefore, it is concluded that strain TRM65318T represents a novel species of the genus of Myceligenerans. The proposed name for this organism is Myceligenerans pegani sp. nov., with type strain TRM65318T (=CCTCC AA 2019057T=LMG 31679T).

Bioprospecting of secondary metabolites produced by endophytic actinomycete isolated from Aloe vera

The aim of this work to evaluate the antibacterial action of secondary metabolites produced by endophytic actinomycetes, against isolates of medical interest. For such, the bacteria were isolated from the botanical sample and classical morphological identification was performed. The induction of metabolites production was carried out using the Submerged Fermentation technique, followed by purification of the extract. The evaluation of antibacterial activity was accomplished by using agar diffusion and Minimum Inhibitory Concentration (MIC) assays. Macro and microscopic analysis of the grown microorganism allowed it to be identified as belonging to the genus Streptomyces. Test results demonstrated that the extract was able to inhibit the growth of C. propinquum 431024, with an inhibition zone of 27±0 mm and of C. propinquum 36063, with an inhibition zone of 24±0 mm and MIC of 32.1 µg/mL for boths. The evaluation of biofilm formation resulted in: C. propinquun 421024 as a weak producer and C. propinquun 36063 as a moderate producer. The biofilm interference test showed that compared to the control there was a reduction in biofilm formation of 22% for C. propinquun 421024, as well as a reduction in pre-formed biofilm of 27%. There was a 48% reduction in the formation and 42% in the pre-formed biofilm of C. propinquun 36063. Lastly, it was concluded that the secondary metabolites produced by the endophytic actinomycetes demonstrated an antibiotic potential against clinical strains (nasopharynx) of Corynebacterium propinquum, a rare pathogen described for causing infections in patients with pulmonary involvement, pleural effusion, bacteremia and endocarditis and which presents an emerging antibiotic resistance.

Enhancing tomato growth with an endophytic actinomycete and applied planting media as its carriers

Enhancing tomato growth with an endophytic actinomycete and applied planting media as its carriers

Isolation and molecular detection of endophytic actinomycetes Nocardiopsis dassonvillei DMS 1 (MH900216) from marine sea grasses with bacterial inactivation

In recent years, new antibiotics have been discovered around the world in order to inhibit multi-drug resistant (MDR) pathogens. To overcome this problem, marine actinomycetes are an alternative choice for producing new bioactive compounds that inhibit MDR bacteria. The typical endophytic actinomycete (EA) Nocardiopsis dassonvillei (N. dassonvillei) DMS 1 (MH900216) was isolated from marine Sea grasses by surface sterilization method. After surface sterilization, it was confirmed that the pure, dry, white-colored spore producing colonies emerged from the internal tissue of the Sea grasses. The crude extract of N. dassonvillei DMS 1 (MH900216) demonstrated 8- and 10-mm zones of inhibition against A. baumannii and K. pneumoniae, respectively. The composition of N. dassonvillei DMS 1 (MH900216) with potential anti-bacterial properties was studied by GC-MS analysis and exhibited 22 chemical compounds. Subsequently, the molecular identification and phylogenetic construction of the isolated EA strain was confirmed as N. dassonvillei DMS 1 (MH900216). The liquid-liquid extraction of the compound demonstrated 24- and 26-mm zones of inhibition against A. baumannii and K. pneumoniae, respectively. Furthermore, the purified crude compound demonstrated 92% and 94% cell death against A. baumannii and K. pneumoniae, respectively, at a minimum inhibitory concentration of 500 μg/mL. Overall, the present study demonstrated the antibacterial properties of the EA N. dassonvillei DMS 1 (MH900216) isolated from Sea grasses and their importance as alternative sources for discovering new antibiotics to inhibit MDR bacteria.

Microbial mediated Synthesis and characterization of Ag Nanoparticles and its Synergistic impact on Actinobacterial, Anticancer and Antihelminthic activity

In this study, an endophytic actinomycetes strain of Streptomyces viridodiastaticus strain MGR23 was isolated from the roots of the medicinal plant Centella asiatica. The biosynthesized Ag-NPs were characterized using UV-Vis spectroscopy, Fourier transform infrared (FT-IR), Scanning electron microscopy (SEM), X-ray diffraction (XRD) and Zeta potential, which confirmed the successful formation of crystalline, spherical silver nanoparticles. Biosynthesized AgNPs significantly inhibited and reduced the growth of medically important survival microorganisms (Streptomyces viridodiastaticus MGR 23 against Streptomyces toxytricini D2, Acidothermus cellulolyticus CA16, Streptomyces sp. MGR 21, Aciditerrimonas ferrireducens SM7 and Streptomyces tritici D5). The biosynthesized Ag-NPs exhibited broad-spectrum antihelminthic activity. Our data elucidated that the biosynthesized Ag-NPs had a highly cytotoxic effect against the cancerous L6 cell line under in vitro conditions, revealing a maximum IC50 value of 1.0 μg/mL and no known anticancer drug was used as a positive regulation. AgNPs have a remarkable synergistic impact that points to their considerable potential in nanomedicine for future clinical use as a combination therapy.

Biocontrol property of Streptomyces parvulus VRR3 in green gram plant (Vigna radiata L.) against Fusarium solani in greenhouse

Actinomycetes are isolated from various habitats including, plants, rhizospheres of.plant roots, terrestrial environment, and marine sediments. These Actinomycetes produce various secondary metabolites with antifungal, antibacterial, anticancer and antidiabetic properties. In the present study, a total of 31 endophytic Actinomycetes strains were isolated from the root of Vigna radiata L. (green gram plant). The antagonistic property of all strains was tested in vitro against Fusarium solani. The mycelia growth inhibition varied between 10.5 ± 1.4 and 80.5 ± 4.2% by Actinomycetes, whereas, the crude extract showed 14.2 ± 1.1 and 78.4 ± 3.1% growth inhibition (p < 0.01). The selected twelve Actinomycetes produced extracellular lytic enzymes. The results revealed that Streptomyces parvulus produced protease (18 ± 2 U/mL), amylase (20 ± 1 U/mL), lipase (13 ± 1 U/mL) and cellulase (8 ± 0 U/mL) at high levels than other Actinomycetes (p < 0.01). In addition, the ability of the screened Actinomycetes to produce plant growth-promoting agents was analyzed. The results showed that S. parvulus and other strains produced Indole acetic acid (1.5 ± 0.3 to 34.1 ± 3.4 μg/mL), phosphate solubilisation ability (1.5–24.3 PO4−3 μg/mL). The thermostable and pH-stable secondary metabolite-producing organism was characterized as S. parvulus VRR3. The in vivo growth-promoting activity of the green gram plant was tested in the greenhouse. The results showed that the selected S. parvulus was able to improve the growth of green gram seedlings, showing enhanced shoot-, and root length (p < 0.01). In addition, the seedlings treated with S. parvulus showed a decreased incidence of Fusarium wilt disease symptoms, whereas the positive control developed >80% disease symptoms (p < 0.01). Hence, S. parvulus VRR3 has the potential to become an important biocontrol agent in modern agricultural practice.

Exogenous application of antagonistic Streptomyces sp. SND-2 triggers defense response in Vigna radiata (L.) R. Wilczek (mung bean) against anthracnose infection

Anthracnose is a devastating disease caused by the fungus Colletotrichum lindemuthianum (CL) in Vigna radiata (L.) R. Wilczek (mung bean). In the present study, an eco-friendly approach to control anthracnose infection, growth promotion and enhancement of defense response in mung bean plants using endophytic actinomycetes was performed. Among the 24 actinomycetes isolates from the Cleome rutidosperma plant, the isolate SND-2 exhibited a broad spectrum of antagonistic activity with 63.27% of inhibition against CL in the dual culture method. Further, the isolate SND-2 was identified as Streptomyces sp. strain SND-2 (SND-2) through the 16S rRNA gene sequence. In-vitro screening of plant growth trials confirmed that SND-2 has the potential to produce indole acetic acid, hydrogen cyanide, ammonia, phosphate solubilization, and siderophore. The in-vivo biocontrol study was performed with exogenous application of wettable talcum-based formulation of SND-2 strain to mitigate CL infection in mung bean seedlings. The results displayed maximum seed germination, vigor index, increased growth parameters, and lowest disease severity (43.63 ± 0.73) in formulation treated and pathogen challenged mung bean plants. Further, the application of SND-2 formulation with pathogen witnessed increased cellular defense through the maximum accumulation of lignin, hydrogen peroxide and phenol deposition in mung bean leaves compared with control treatments. Biochemical defense response exhibited upregulation of antioxidant enzymes such as phenylalanine ammonia-lyase, β-1,-3-Glucanase, and peroxidase enzymes activities with increased phenolic (3.64 ± 0.11 mg/g fresh weight) and flavonoid (1.14 ± 0.05 mg/g fresh weight) contents in comparison with other treatments at 0, 4, 12, 24, 36, and 72 h post pathogen inoculation. This study demonstrated that formulation of Streptomyces sp. strain SND-2 is a potential source as a suppressive agent and plant growth promoter in mung bean plants upon C. lindemuthianum infestation and witnesses the elevation in cellular and biochemical defense against anthracnose disease.

Diversity analysis of endophytes with antimicrobial and antioxidant potential from Viola odorata: an endemic plant species of the Himalayas.

Viola odorata, also known as "Banafshah" in high altitudes of Himalayas, is well known for its pharmaceutical importance in Ayurvedic and Unani medicinal system. The plant is a source of various drugs for its anti-inflammatory, diaphoretic, diuretic, emollient, expectorant, antipyretic, and laxative properties. The endophytes of plants have been reported for their role in modulating various physiological and biological processes of the host plants. In the present study, a total of 244 endophytes were isolated in pure cultures from the roots of Viola odorata, and genetic diversity was evaluated using amplified ribosomal DNA restriction analysis (ARDRA) and enterobacterial repetitive intergenic consensus (ERIC). The molecular fingerprinting revealed variation among various rRNA types among morphologically different endophytes based on ARDRA and ERIC-PCR. The screening of endophytes showed antimicrobial activity of 11 bacterial isolates and one actinomycete SGA9 against various pathogens Bacillus cereus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. The antioxidant activity revealed the majority of the bacterial isolates able to scavenge the free radical in the range of 10-50% and 8 bacterial isolates in the range of 50-85%. Principal component analysis separated eight isolates away from the central eclipse and form a separate group based on antimicrobial and antioxidant potential. The identification of these eight isolates showed affiliation with different species of the genus Enterobacter, Microbacterium, Pseudomonas, Rhizobium, and Streptomyces. This is the first report on the characterization of endophytic bacteria and actinomycetes from endemic Viola odorata. Results suggested that these endophytes could be explored for the production of antimicrobial and antioxidant products.

Development of a drug discovery approach from microbes with a special focus on isolation sources and taxonomy.

After the successful discoveries of numerous antibiotics from microorganisms, frequent reisolation of known compounds becomes an obstacle in further development of new drugs from natural products. Exploration of biological sources that can provide novel scaffolds is thus an urgent matter in drug lead screening. As an alternative source to the conventionally used soil microorganisms, we selected endophytic actinomycetes, marine actinomycetes, and actinomycetes in tropical areas for investigation and found an array of new bioactive compounds. Furthermore, based on the analysis of the distribution pattern of biosynthetic gene clusters in bacteria together with available genomic data, we speculated that biosynthetic gene clusters for secondary metabolites are specific to each genus. Based on this assumption, we investigated actinomycetal and marine bacterial genera from which no compounds have been reported, which led to the discovery of a variety of skeletally novel bioactive compounds. These findings suggest that consideration of environmental factor and taxonomic position is critically effective in the selection of potential strains producing structurally unique compounds.

Mumia quercus sp. nov., isolated from the root of Quercus variabilis.

A novel endophytic actinomycete, designated strain NEAU-365T, was isolated from the root of Quercus variabilis collected from Nanjing, Jiangsu, PR China. Comparative 16S rRNA gene sequencing showed that strain NEAU-365T belonged to the genus Mumia but clearly differs from the currently recognized species Mumia zhuanghuii Z350T (99.31 %), Mumia xiangluensis NEAU-KD1T (98.82 %) and Mumia flava MUSC 201T (97.78 %). Phylogenetic tree analysis revealed that strain NEAU-365T clustered with the type strains of the genus Mumia. The genome size was 4.1 Mbp with a DNA G+C content of 71.2 mol%. Digital DNA-DNA hybridization and average nucleotide identity values between the genome sequence of strain NEAU-365T and those of M. zhuanghuii Z350T(27.5 and 84.0 %), M. xiangluensis NEAU-KD1T(23.4 and 80.9 %) and M. flava MUSC 201T(20.9 and 77.7 %) were below the recommended thresholds for species delineation. Cells were observed to be irregular cocci shaped. The cell wall contained ll-diaminopimelic acid and the whole-cell sugars were galactose and rhamnose. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid and three unidentified phospholipids. The predominant menaquinone was detected as MK-9(H4). The major fatty acids were C16 : 0, C18 : 0, C18 : 1 ω9c and 10-methyl C18 : 0. On the basis of genotypic and phenotypic differences from members of the genus Mumia, a novel species, Mumia quercus sp. nov. is proposed. The type strain is NEAU-365T (=CCTCC AA 2021033T=JCM 35005T).

Identification and genomic analyses of a novel endophytic actinobacterium Streptomyces endophytica sp. nov. with potential for biocontrol of yam anthracnose.

Anthracnose disease caused by Colletotrichum gloeosporioides is one of the devastating diseases of yams (Dioscorea sp.) worldwide. In this study, we aimed to isolate endophytic actinobacteria from yam plants and to evaluate their potential for the control of yam anthracnose based on bioassays and genomic analyses. A total of 116 endophytic actinomycete strains were isolated from the surface-sterilized yam tissues from a yam orchard in Hainan Province, China. In total, 23 isolates showed antagonistic activity against C. gloeosporioides. An endophytic actinomycete, designated HNM0140T, which exhibited strong antifungal activities, multiple biocontrol, and plant growth-promoting (PGP) traits was subsequently selected to colonize in the tissue-cultured seedlings of yam and was tested for its in vivo biocontrol potential on yam anthracnose. The results showed that treatment with strain HNM0140T markedly reduced the severity and incidence of yam anthracnose under greenhouse conditions. Morphological and chemotaxonomic analyses showed that strain HNM0140T was assigned to the genus Streptomyces. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain HNM0140T formed a separate cluster together with Streptomyces lydicus ATCC 25470T (99.45%), Streptomyces chattanoogensis NRRL ISP-5002T (99.45%), and Streptomyces kronopolitis NEAU-ML8T (98.97%). The phylogenomic tree also showed that strain HNM0140T stably clustered with Streptomyces lydicus ATCC 25470T. The ANI and dDDH between strain HNM0140T and its closest related-type species were well below the recommended thresholds for species demarcation. Hence, based on the phylogenetic, genomic, and phenotypic analyses, strain HNM0140T should represent a new streptomycete species named Streptomyces endophytica sp. nov. Genomic analysis revealed that strain HNM0140T harbored 18 putative BGCs for secondary metabolites, some PGP-related genes, and several genes coding for antifungal enzymes. The presented results indicated that strain HNM0140T was a promising biocontrol agent for yam anthracnose.