Multi-Omics Analysis Demonstrates the Critical Role of Non-Ethanolic Components of Alcoholic Beverages in the Host Microbiome and Metabolome: A Human- and Animal-Based Study.

It is known that alcoholic beverages alter the human gut microbiome. This study focused on the potential impact of non-ethanolic ingredients in whisky on the gut bacteriome. A pilot study was carried out on 15 whisky drinkers, 5 rice beer drinkers, and 9 non-drinkers to determine the effect of alcoholic beverages on the host microbiome and metabolome. Additionally, a mouse model was used to assess the differential impact of three whisky brands (each with an equal ethanol concentration). The results indicate that the non-ethanolic components have an impact on the gut microbiome, as well as on the metabolites in blood and feces. The amount of Prevotella copri, a typical core Indian gut bacterium, decreased in both the human and mouse groups of whisky type 1, but an increase in abundance of Helicobacteriaceae (p = 0.01) was noticed in both groups. Additionally, the alcohol-treated cohorts had lower levels of short-chain fatty acids (SCFAs), specifically butyric acid, and higher amounts of lipids and stress marker IL1-ß than the untreated groups (p = 0.04-0.01). Furthermore, two compounds, ethanal/acetaldehyde (found in all the whisky samples) and arabitol (unique to whisky type 1), were tested in the mice. Similar to the human subjects, the whisky type 1 treated mouse cohort and the arabitol-treated group showed decreased levels of Prevotella copri (p = 0.01) in their gut. The results showed that non-ethanolic compounds have a significant impact on host gut bacterial diversity and metabolite composition, which has a further vital impact on host health. Our work further emphasizes the need to study the impact of non-ethanolic ingredients of alcoholic beverages on host health.

Wound healing efficacy of rhamnolipid-coated zinc oxide nanoparticle along with its in vivo antibacterial efficacy against Staphylococcus aureus.

Rhamnolipids are microbial metabolites with antibacterial efficacies, which can be further boosted through the application of nanobiotechnology. In this study, the efficacy of rhamnolipid-coated zinc oxide nanoparticles (ZnRL) has been studied for their wound healing efficacy as well as in vivo antibacterial efficacy. Thus, this study evaluates the efficacy of ZnRL to heal an excised infected wound, which was compared with the healing efficacy of rhamnolipid and clindamycin. The study revealed that rhamnolipid-coated zinc oxide nanoparticles possess promising wound healing efficacy with prominent antibacterial activity in the rat model. Prominent wound healing in a Staphylococcus aureus infected excised wound was observed on the 5th day of the treatment when the wound site was treated with 100 µl of 0.5 mg/ml of ZnRL. This concentration of ZnRL was found to exhibit efficient antibacterial activity against the pathogen, thereby decreasing the amount of pathogen in the wound site. ZnRL exhibited efficient wound contraction, thereby decreasing the size of the wound prominently in 5 days. Histological study revealed efficient tissue remodelling in ZnRL-treated skin which resulted in rapid formation of the epidermis and recruitment of various dermal cells within the 5th day of treatment. The study also revealed the non-cytotoxic effect of the nanoparticles in fibroblast cell line L929 and the non-haemolytic effect against blood cells, indicating its potential in pharmaceuticals.

Green synthesized Se nanoparticle-mediated alleviation of salt stress in field mustard, TS-36 variety.

During this decade, selenium nanoparticles have been found to play a crucial role in helping plants endure several stress conditions, which thereby helps enhance the production of crops in such harsh environments. Globally, high salinity is considered a long-term stress in the crop fields which affects the growth and production of many crops, including mustard-one of the most important oil crops. Here, the activities of spherical-shaped selenium nanoparticles with an average particle size of 55.81 nm, synthesized and functionalized by phytochemicals of fresh grape aqueous extract, were evaluated in the salinity stress (200 mM NaCl) tolerance of mustard plants grown hydroponically in modified Hoagland's solution. These bioactive nanoparticles (30 mg L-1) have exhibited significant activity in alleviating the salt stress complications in mustard, enhancing the activities of antioxidant enzymes (SOD 41.20 %, CAT 64.10 %, APX 63.06 %, and POX 70.43 %), phenolic content (98.88 %), flavonoid content (86.90 %), and free radical scavenging activity (61.89 %). The seed germination percentage, root and shoot length, fresh and dry weight per plant, water content percentage, chlorophyll content, carbohydrate content, and protein content were significantly improved by 39.66 %, 75 %, 60.64 %, 41.2 %, 22.11 %, 1.02 %, 81.92 %, 24.65 % and 79.14 % respectively by the nano selenium application during NaCl stress compared to the control group growing under salt stress without nanoparticles. Gas chromatography-mass spectrometry chromatogram analysis inferred the interaction between the nano-selenium and mustard plants under salt stress. Besides, the in-silico analysis revealed the active molecular interactions between selenium and 20 different proteins of mustard, including glutathione peroxidase, an important antioxidant enzyme.

Se nanoparticles stabilized with Allamanda cathartica L. flower extract inhibited phytopathogens and promoted mustard growth under salt stress.

Selenium Nanoparticles (SeNPs) exhibit tremendous application in agriculture as antimicrobials or as nano fertilizer. Present work reports the eco-friendly synthesis of SeNPs by using Allamanda cathartica L. flower extract (aqueous) as a reducing/capping agent and selenium dioxide as a precursor. The method used here is free of any toxic reducing agents and organic solvents. The synthesis process of SeNPs took 5 h at 60 °C, confirmed by the brick red colour of the solution followed by UV-Vis spectroscopy and further characterized by XRD, FTIR, EDX and SEM. The average size (diameter) of the SeNPs were found to be 60.31 nm by DLS. It has shown strong antimicrobial activity against Pseudomonas marginalis and P. aeruginosa at 2.5, 5 and 10 mg/mL concentrations. Besides, its application improved seed germination and growth parameters of Brassica campestris (TS 36 variety) under salt stress. 25 mg/L SeNPs has improved the germination percentage by around 31%, shoot length by 92%, root length by 78% and total chlorophyll content by 49% under 200 mM NaCl stress. This SeNPs could be a potential antimicrobial agent in treating plant diseases caused by the mentioned phytopathogens, having no or minimum toxicity, in fact having positive impacts on plant growth.

Biodegradation of hazardous naphthalene and cleaner production of rhamnolipids - Green approaches of pollution mitigation.

Toxic and hazardous waste poses a serious threat to human health and the environment. Green remediation technologies are required to manage such waste materials, which is a demanding and difficult task. Here, effort was made to explore the role of Pseudomonas aeruginosa SR17 in alleviating naphthalene via catabolism and simultaneously producing biosurfactant. The results showed up to 89.2% naphthalene degradation at 35 °C and pH 7. The GC/MS analysis revealed the generation of naphthalene degradation intermediates. Biosurfactant production led to the reduction of surface tension of the culture medium to 34.5 mN/m. The biosurfactant was further characterized as rhamnolipids. LC-MS of the column purified biosurfactant revealed the presence of both mono and di rhamnolipid congeners. Rhamnolipid find tremendous application in medical field and as well as in detergent industry and since they are of biological origin, they can be used as favorable alternative against their chemical counterparts. The study demonstrated that catabolism of naphthalene and concurrent formation of rhamnolipid can result in a dual activity process, namely environmental cleanup and production of a valuable microbial metabolite. Additionally, the present-day application of rhamnolipids is highlighted.

Synthesis, characterization, and evaluation of antibacterial efficacy of rhamnolipid-coated zinc oxide nanoparticles against Staphylococcus aureus.

The antimicrobial efficacy of rhamnolipid is well established against a wide range of pathogens. However little is known about the enhancement of antimicrobial efficacy of rhamnolipid in the form of nanoparticles. With a curiosity of enhancing antimicrobial activity, a study has been carried out to evaluate the antimicrobial efficacy of rhamnolipid-coated zinc oxide nanoparticles. The zinc oxide nanoparticles were synthesized with rhamnolipid, produced by Pseudomonas aeruginosa JS29. The rhamnolipid-coated zinc oxide nanoparticles were characterized by FTIR, XRD, TGA, TEM, and SAED. The antimicrobial and antibiofilm efficacy of the nanoparticles was evaluated against Staphylococcus aureus MTCC 96. FTIR, XRD, TEM, and SAED analyses confirmed that the nanoparticles contain both rhamnolipid and zinc as constituents and are polycrystalline with sizes ranging from 40 to 50 nm. At a concentration of 250 µg/ml, rhamnolipid-coated zinc oxide nanoparticles exhibited 80% growth inhibition of the pathogen. Again, at the same concentration, the nanoparticle was observed to inhibit 78% of biofilm formation while disrupting 100% of preformed biofilm. The nanoparticles demonstrated an enhanced inhibitory and antibiofilm efficacy against the pathogen compared to the individual effect of both rhamnolipid and zinc oxide nanoparticles. With the established non-toxicity of rhamnolipid-coated zinc oxide nanoparticles in fibroblast cell lines, the nanoparticles could be a promising pharmaceutical alternative.

Densitometric analysis of rep-PCR data: Insight into genetic variability and transmission of Clostridium perfringens typed with an improved multiplex PCR.

An epidemiological study was conducted in North-East India (part of Indo-Burma biodiversity hotspot) to better understand the distribution, diversity, and transmission of Clostridium perfringens among livestock, pets, wild animals (captive), and humans. A total of 160 C. perfringens isolates were recovered from 642 diarrhoeic faecal samples with an isolation rate of 24.92%. Isolation rate was the highest among captive wild animals (37.5%) followed by dog (34.6%), human (33.8%), pig (32.7%), cattle (20.8%), goat (18.3%) and poultry (9.3%). Isolates were toxin typed using a seven gene multiplex PCR designed for simultaneous detection of cpa, cpb, cpb2, etx, iap, cpe and netB. The majority of isolates, 128 (80%) were of type A, followed by 17 (10.62%), 5 (3.12%), 4 (2.5%), 3 (1.87%), 2 (1.25%) and 1 (0.63%) isolates of type C, D, E, G, F and B, respectively. Beta 2 toxin gene was present in 65 (50%) of type A isolates, followed by 7 (41.2%), 4 (80%), 1(25%), and 1 (100%) of type C, D, G and B isolates, respectively. Beta 2 toxin has a high prevalence among dogs (28.6%), cattle (27.3%), and pig (20.8%) compared to humans, goat, wild animals, and poultry (1.2-14.3%). The prevalence of CPE and NetB toxin-positive strains was low, with only 3 (1.8%) and 5 (3.1%) isolates, respectively. Association of C. perfringens with diarrhoea in Civet Cat, Golden Langur, and Gray Langur has been reported for the first time. The genetic diversity and transmission of isolates were investigated using automated rep-PCR (Diversilab®, bioMérieux) using two densitometry-based matrices: modified Kullback-Leibler (KL) and Pearson's correlation (PC). The PC and modified KL matrices formed three distinct clusters with 59% and 27.2% similarity, respectively. C. perfringens diversity and transmission were best studied using modified KL matrix that placed more emphasis on the presence of bands rather than intensity. However, the PC method was found to be more suitable for differentiating strains within a toxin type, with slightly higher D-values.

Effects of Resveratrol and Mangiferin on PPARγ and FALDH Gene Expressions in Adipose Tissue of Streptozotocin-Nicotinamide-Induced Diabetes in Rats.

Type 2 diabetes (T2D) is characterized by insufficient insulin secretion by the pancreatic beta cells and insulin resistance in liver, skeletal muscle, and white adipose tissue. Adipose tissue plays a major role in glucose homeostasis and lipid metabolism. Dietary antioxidants such as resveratrol and mangiferin may offer some protection against the early stage of diabetes mellitus. Therefore, an attempt has been made to investigate the effects of resveratrol and mangiferin on biochemical parameters and molecular mechanism of PPARγ and FALDH gene expression in adipose tissue of streptozotocin- (STZ-) nicotinamide- (NA-) induced diabetic rats. Albino Wister rats were randomly divided into five groups: control rats (Group 1), diabetic control rats (Group 2), diabetic rats given resveratrol (40 mg/kg body weight per day; Group 3), diabetic rats given mangiferin (40 mg/kg body weight per day; Group 4), diabetic rats given glibenclamide (0.6 mg/kg body weight per day; Group 5). Serum biochemical parameters-total cholesterol (TC), total triglyceride (TG), low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, glycosylated hemoglobin (HbA1c), urea, and uric acid were analyzed. We found that the oral administration of resveratrol and mangiferin to STZ-NA-induced diabetic rats for 30 days showed the significant protective effect on all the biochemical parameters. A significant reduction in blood glucose and HbA1c levels was observed in rats treated with 40 mg/kg body weight per day of resveratrol or mangiferin. Moreover, both these antioxidants showed significant enhancement of PPARγ and FALDH gene expression in rat adipose tissue compared to control rats.

Draft Genome Sequence of Bacillus altitudinis Lc5, a Biocontrol and Plant Growth-Promoting Endophyte Strain Isolated from Indigenous Black Rice of Manipur.

We report here the 3.6-Mb draft genome of Bacillus altitudinis Lc5, a potential plant growth promoter and an active antagonistic endophyte of black rice. This genome study will provide better insights into the strain's mechanisms for plant growth promotion and biocontrol, thus facilitating its application in organic agriculture.

Withdrawal from high-carbohydrate, high-saturated-fat diet changes saturated fat distribution and improves hepatic low-density-lipoprotein receptor expression to ameliorate metabolic syndrome in rats.

OBJECTIVE: The "lipid hypothesis" determined that saturated fatty acid (SFA) raises low-density lipoprotein cholesterol, thereby increasing the risk for metabolic syndrome (MetS). The aim of this study was to investigate the effect of subchronic withdrawal from a high-carbohydrate, high-saturated fat (HCHF) diet during MetS with reference to changes in deleterious SFA (C12:0, lauric acid; C14:0, myristic acid; C16:0, palmitic acid; C18:0, stearic acid) distribution in liver, white adipose tissue (WAT), and feces. METHODS: MetS induced by prolonged feeding of an HCHF diet in Wistar albino rat is used as a model of human MetS. The MetS-induced rats were withdrawn from the HCHF diet and changed to a basal diet for final 4 wk of the total experimental duration of 16 wk. SFA distribution in target tissues and hepatic low-density lipoprotein receptor (LDLr) expression were analyzed. RESULTS: Analyses of changes in SFA concentration of target tissues indicate that C16:0 and C18:0 reduced in WAT and liver after withdrawal of the HCHF diet. There was a significant (P < 0.001) decrease in fecal C12:0 with HCHF feeding, which significantly (P < 0.01) increased after withdrawal of this diet. Also, an improvement in expression of hepatic LDLr was observed after withdrawal of HCHF diet. CONCLUSION: The prolonged consumption of an HCHF diet leads to increased SFA accumulation in liver and WAT, decreased SFA excretion, and reduced hepatic LDLr expression during MetS, which is prominently reversed after subchronic withdrawal of the HCHF diet. This can contribute to better understanding of the metabolic fate of dietary SFA during MetS and may apply to the potential reversal of complications by the simple approach of nutritional modification.

Uptake of Total Petroleum Hydrocarbon (TPH) and Polycyclic Aromatic Hydrocarbons (PAHs) by Oryza sativa L. Grown in Soil Contaminated with Crude Oil.

The purpose of this study was to determine whether total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbons (PAHs) present in crude oil contaminated sites are transferred to roots, shoots and finally the grains of rice crops (Oryza sativa L.) grown in those sites. Soil was artificially contaminated with crude oil at concentrations of 0, 1000, 5000, 10,000, and 15,000 mg/kg, followed by planting of rice seedlings. After harvest, TPH in plant samples were measured, and it was determined that the uptake of TPH by the plants gradually increased as the concentration of oil in soil increased. Further, from GC-MS analysis, it was observed that PAHs including naphthalene and phenanthrene bioaccumulated in rice plant parts. Vital physico-chemical properties of soil were also altered due to crude oil contamination. Our study revealed that rice plants grown in crude oil polluted sites can uptake TPH including PAHs, thus emphasising the importance of prior investigation of soil condition before cultivation of crops.

Utilization of Paneer Whey Waste for Cost-Effective Production of Rhamnolipid Biosurfactant.

The present study aimed at isolating rhamnolipid biosurfactant-producing bacteria that could utilize paneer whey, an abundant waste source as sole medium for the production purpose. Pseudomonas aeruginosa strain, SR17, was isolated from hydrocarbon-contaminated soil that could efficiently utilize paneer whey for rhamnolipid production and reduce surface tension of the medium from 52 to 26.5 mN/m. The yield of biosurfactant obtained was 2.7 g/l, upgraded to 4.8 g/l when supplemented with 2 % glucose and mineral salts. Biochemical, FTIR, and LC-MS analysis revealed that extracted biosurfactant is a combination of both mono and di-rhamnolipid congeners. The critical micelle concentration (CMC) was measured to be 110 mg/l. Emulsification activity of the biosurfactant against n-hexadecane, olive oil, kerosene, diesel oil, engine oil, and crude oil were found to be 83, 88, 81, 92, 86, and 100 %, respectively. The rhamnolipid was detected to be non-toxic against mouse fibroblastic cell line L292.

Identification and characterization of Fusarium sp. using ITS and RAPD causing fusarium wilt of tomato isolated from Assam, North East India.

Fusarium oxysporum which causes wilt is a serious pathogen. Fusarium isolates were isolated from Assam located in North East region of India. Morphological identification of Fusarium isolates was done using conidial and hyphal structures. Molecular identification of Fusarium isolates was done by amplifying the internal transcribed spacer (ITS) region of the conserved ribosomal DNA using primers ITS1 and ITS4. All the ITS sequences were compared for gaps and similarity. Further, characterization of random amplified polymorphic DNA (RAPD) was carried out using 40 primers. 15 primers that gave reproducible results were selected. RAPD was used to observe the relatedness among these isolates. Thus, it was concluded that molecular profiling using ITS is an indispensable method for identification studies.

Chrysin rich Scutellaria discolor Colebr. induces cervical cancer cell death via the induction of cell cycle arrest and caspase-dependent apoptosis.

AIMS: Scutellaria discolor Colebr. has been extensively used in traditional medicine against several diseases. The purpose of this study was to investigate the anticancer potential of S. discolor and to isolate the bioactive principle responsible for the anticancer activity. METHODS: Cytotoxicity experiments were performed on cancer and normal cells using MTT assay. The mechanism of cell death was evaluated using real time PCR array, fluorescence microscopy, flow cytometry and Western blotting. MTT assay guided isolation (partition and column chromatography) was performed to identify the antiproliferative principle. Quantification of the active principle was done using HPLC. KEY FINDINGS: Acetone extract of S. discolor (SDE) inhibited the growth and survival of cancer cells to varying degree, but the inhibition was found to be maximum in cervical cancer cell lines. There was no significant toxicity induced to normal cells. The cell death was mediated through apoptosis. There was increased mitochondrial membrane depolarization, expression of Bax, caspase-9, caspase-3 and cleaved-PARP indicating that SDE-induced caspase dependent apoptosis in HeLa cells. Moreover, SDE caused cell cycle arrest in G2 phase in HeLa cells. Cytotoxicity guided fractionation of SDE led to the isolation of chrysin as the active principle responsible for the antiproliferative activity for cervical cancer cells. Interestingly, chrysin was the major phytochemical constituent present in S. discolor. SIGNIFICANCE: S. discolor is an important anticancer plant and a new source of chrysin.

Cephalotaxus griffithii Hook.f. needle extract induces cell cycle arrest, apoptosis and suppression of hTERT and hTR expression on human breast cancer cells.

BACKGROUND: Cephalotaxus spp. are known to possess anticancer potential. In this present work, for the first time the effects of C. griffithii needle (CGN) extracts on human cancer cells were examined. METHODS: The CGN was successively extracted with petroleum ether (PE), acetone and methanol. The extracts were tested for its effect on proliferation of cancer cells (MTT assay on HeLa, ZR751 and HepG2). Extract that showed the maximum growth inhibitory effect was subjected for mechanism of action study. These included apoptosis (morphological and DNA fragmentation assay), cell cycle (flow cytometry), caspase expression (Western blot) and activity (assay kit), p53 (western blot and TP53 siRNA interference) and telomerase expression (reverse transcriptase PCR) analysis. RESULTS: Among the extracts, PE extract induced maximum cytotoxicity, with highest death occurred in ZR751 cells. Since, PE extract induced cell death was highest among the CGN extracts, with maximum cancer cell death occurred in ZR751 cells; we carried out mechanism study of PE extract induced ZR751 cell death. It was observed that PE extract induced ZR751 cell death was associated with cell cycle arrest and apoptosis by activating both intrinsic and extrinsic apoptotic pathways. Knock down study revealed that p53 is essential for loss of ZR751 cell viability induced by PE extract. Further, PE extract down-regulated hTERT, hTR, and c-Myc expression. Thin layer chromatography analysis indicated the presence of unique phytochemicals in PE extract. CONCLUSION: Based on the observations, we concluded that PE extract of C. griffithii needle contains important phyto-components with multiple cellular targets for control of breast cancer and is worthy of future studies.

HPLC analysis of harringtonine and homoharringtonine in the needles of Cephalotaxus griffithii alkaloid fraction and cytotoxic activity on chronic myelogenous leukaemia K562 cell.

Harringtonine (HT) and homoharringtonine (HHT) are Cephalotaxus alkaloids with considerable antileukaemic activity. The objectives of this research were to (1) determine the content of HT and HHT present in Cephalotaxus griffithii needles alkaloid fraction (CGAF) and (2) compare the antiproliferative activity of CGAF, with that of HT and HHT on chronic myelogenous leukaemia K562 cell. The concentration of HT and HHT was found to be 122.14 and 16.79 mg/g of CGAF, respectively. Treatment of K562 cells with CGAF, HT and HHT decreased the viable cells in a dose- and time-dependent manner. Interestingly, the maximum cell death was found in CGAF, with IC50 value which was 3- to 4.6-fold lower than those of HT and HHT. Our results indicate that HT content in the needles of C. griffithii is higher than HHT, and alkaloids other than HT and HHT in CGAF are predominantly responsible for K562 cell death.

Isolation, screening, characterization, and selection of superior rhizobacterial strains as bioinoculants for seedling emergence and growth promotion of Mandarin orange (Citrus reticulata Blanco).

Mandarin orange (MO) is an important fruit crop of tropical and subtropical regions of the world. A total of 217 morphologically distinct rhizobacteria from MO orchards in 3 states of northeastern India were isolated and analyzed for 4 plant-growth-promoting (PGP) attributes: nitrogen fixation, production of indole acetic acid like substances, solubilization of phosphate, and ability to antagonize pathogenic fungi. Isolates were ranked based on in-vitro-assayed PGP attributes, and 10 superior isolates were selected to test their effect on seedling emergence and seedling growth in a completely randomized pot experiment. These 10 isolates increased seedling emergence over a noninoculated control within 45 days after sowing. Five isolates, namely RCE1, RCE2, RCE3, RCE5, and RCE7, significantly increased shoot length, shoot dry biomass, and root dry biomass of 120-day-old seedlings over the noninoculated control. The beneficial effects of 4 selected strains, namely Enterobacter hormaechei RCE-1, Enterobacter asburiae RCE-2, Enterobacter ludwigii RCE-5, and Klebsiella pneumoniae RCE-7, on growth of the seedlings were visible up to 1 year after their transfer to 8 kg capacity pots. These strains were superior both in terms of in-vitro-assayed PGP attributes and of their beneficial effect in low phosphorus soil and, thus, may be promising bioinoculants for promoting early emergence and growth of MO seedlings.

Characterization of the biofuel potential of a newly isolated strain of the microalga Botryococcus braunii Kützing from Assam, India.

Botryococcus braunii GUBIOTJTBB1 was isolated from a freshwater reservoir in Assam, India and its taxonomic identity was confirmed by 18S rRNA sequence analysis. Biofuel potential of the microalga strain was assessed from batch culture under laboratory conditions, based on its lipid content and energy value of the dried biomass. Total lipid of 57.14% and hexane extractable crude hydrocarbon of 52.6% were recorded maximum at 56 and 28 days respectively, which vary upon culture durations. The energy value (54.69 kJ/g) of the strain's sundried biomass was found higher than that of petroleum diesel fuel and nearly twice than other microalgae strains compared. The strain GUBIOTJTBB1 was found superior in terms of total lipid and hydrocarbon contents comparing to the previously reported Indian strains of B. braunii. With further improvements in growth, the strain could become an ideal feedstock for potential biofuel production in the prevailing climatic conditions of the region.

Agrobacterium-mediated genetic transformation of Pogostemon cablin (Blanco) Benth. Using leaf explants: bactericidal effect of leaf extracts and counteracting strategies.

An optimized protocol for Agrobacterium tumefaciens-mediated transformation of patchouli using leaf disk explants is reported. In vitro antibacterial activity of leaf extracts of the plants revealed Agrobacterium sensitivity to the extracts. Fluorometric assay of bacterial cell viability indicated dose-dependent cytotoxic activity of callus extract against Agrobacterium cells. Addition of 0.1% Tween 20 and 2 g/l L-glutamine to Agrobacterium infection medium counteracted the bactericidal effect and significantly increased the T-DNA delivery to explants. A short preculture of explants for 2 days followed by infection with Agrobacterium in medium containing 150 µM of acetosyringone were found essential for efficient T-DNA delivery. Cocultivation for 3 days at 22 °C in conjunction with other optimized factors resulted in maximum T-DNA delivery. The Agrobacterium-mediated transformation of leaf disk explants were found significantly related to physiological age of the explants, age and origin of the of the donor plant. Leaf explants from second node of the 3-month-old in vivo plants showed highest transformation efficiency (94.3%) revealed by transient GUS expression assay. Plants selected on medium containing 20 mg/l kanamycin showed stable GUS expression in leaves and stem. The elongated shoots readily developed roots on kanamycin-free rooting medium and on transfer to soil, plants were successfully established. Polymerase chain reaction (PCR) and reverse-transcriptase PCR analysis in putative plants confirmed their transgenic nature. The established transformation method should provide new opportunities for the genetic improvement of patchouli for desirable trait.