Antibacterial and biofilm inhibition activity of biofabricated silver nanoparticles against Xanthomonas oryzae pv. oryzae causing blight disease of rice instigates disease suppression.

Antimicrobial activity of silver nanoparticles (AgNPs) has been well documented in earlier studies. As their efficient role in combating phytopathogens has begun recently, there is a huge scope to explore their effectiveness in agriculture. Considering the strong antifungal activity of biosynthesized AgNPs (as reported in our previous study), our main aim is to elucidate their antibacterial activity against bacterial plant pathogens to authenticate their wide range of agricultural applications. The present manuscript highlights the potential role of biosynthesized AgNPs against Xanthomonas oryzae pv. oryzae (Xoo) causing disastrous sheath blight disease of rice worldwide. We observed strong antibacterial activity of biosynthesized AgNPs (size ~ 12 nm) against Xoo at 20, 30 and 50 µg/mL concentrations. The significant inhibitory impact of AgNPs on biofilm formation by Xoo was noted even at the lower dose of 5 µg/mL (p = 0.001). Maximum biofilm inhibition (p = 0.000) was caused at 50 µg/mL concentration of AgNPs in comparison to control. Furthermore, disease suppression by biosynthesized AgNPs was authenticated under greenhouse conditions. Foliar spray of AgNPs significantly reduced the blight symptoms in rice sheaths as shown by 9.25% DLA (% Diseased leaf area) as compared to 33.91% DLA in Xoo inoculated rice plants. Altogether, our data suggest that biosynthesized AgNPs based nanoformulation can be applied for successful management of blight disease of rice. In addition, the antibiofilm strategies instigated by AgNPs can be exploited against a wide range of bacterial phytopathogens. In light of rapidly emerging antibiotic-resistant microbial strains, the current work provides an alternate effective platform for the application of nanoformulation for augmenting sustainability in the agriculture.

Potential of biosynthesized silver nanoparticles using Stenotrophomonas sp. BHU-S7 (MTCC 5978) for management of soil-borne and foliar phytopathogens.

Stenotrophomonas sp. is emerging as a popular microbe of global concern with various potential ecological roles. Biosynthesis of gold and silver nanoparticles (AgNPs) using this bacterial strain has shown promising applications in life sciences. However, there is no report on efficient agricultural applications of biosynthesized AgNPs using Stenotrophomonas sp. In this regard, successful biosynthesis of AgNPs using Stenotrophomonas sp. BHU-S7 (MTCC 5978) was monitored by Uv-visible spectrum showing surface plasmon resonance (SPR) peak at 440 nm. The biosynthesized AgNPs were spherical with an average mean size of ~12 nm. The antifungal efficacy of biosynthesized AgNPs against foliar and soil-borne phytopathogens was observed. The inhibitory impact of AgNPs (2, 4, 10 µg/ml) on conidial germination was recorded under in vitro conditions. Interestingly, sclerotia of Sclerotium rolfsii exposed to AgNPs failed to germinate on PDA medium and in soil system. Moreover, AgNPs treatment successfully managed collar rot of chickpea caused by S. rolfsii under greenhouse conditions. The reduced sclerotia germination, phenolic acids induction, altered lignification and H2O2 production was observed to be the probable mechanisms providing protection to chickpea against S. rolfsii. Our data revealed that AgNPs treated plants are better equipped to cope with pathogen challenge pointing towards their robust applications in plant disease management.

Effect of mixture of Trichoderma isolates on biochemical parameter in tomato fruits against Sclerotinia sclerotiorum rot of tomato plant.

Experiments revealed that a mixture of Trichoderma harzianum isolates, BHU51 and BHU105 showed lowest mean disease rating (MDR) of 1.70 and 1.62% and per cent disease reduction (PDR) by 41.00 and 44.84% during the year 2008-09 and 2009-10, respectively. Shoot length, chlorophyll content and yield was also recorded highest in the mixture of BHU51+ BHU105 treatment followed by single Tichoderma treatments while lowest was found in pathogen inoculated control. The nutritional quality such as lycopene content, protein and carbohydrate was recorded highest in BHU51+ BHU105 treatment. The antioxidant activity and free radical scavenging ability of tomato fruit extract was also recorded. The results indicated that maximum 1,1-diphenyl -2-pycryl-hydrazyl (DPPH) radical scavenging activity (47.86%), ferrous ion chelation capacity (50.81%), hydroxyl radical scavenging ability (49.18%) and reducing power 0.203 O.D. at wavelength 700 nm was maximum for BHU51+ BHU105 treatment, followed by single Trichoderma treated treatments while these were recorded lowest in pathogen inoculated control.

Silver nanoparticles mediated altered gene expression of melanin biosynthesis genes in Bipolaris sorokiniana.

Melanin production in many fungal phytopathogens has been investigated to play direct or indirect role in pathogenesis. However, in Bipolaris sorokiniana, the spot blotch pathogen of wheat, much less is known about the role melanin play in pathogenesis. As an extension of our previous report, the present study aims to investigate the plausible association between melanin production and virulence factor in B. sorokiniana. In the previous study, we carried out analysis on the antifungal efficacy of biosynthesized silver nanoparticles (AgNPs) against B. sorokiniana. The present investigation revealed the gene expression analysis of melanin biosynthesis genes viz. polyketide synthase (PKS1) and scytalone dehydratase (SCD1) under the influence of AgNPs. The 0.05mg/ml concentration of AgNPs yielded noticeable inhibition of B. sorokiniana growth, while 0.1mg/ml concentration of AgNPs accounted for complete inhibition of pathogen growth. In addition, the semiquantitative RT-PCR analysis exhibited reduced expression of PKS1 and SCD1 under the influence of AgNPs treatment. Furthermore, the qRT-PCR demonstrated 6.47 and 1.808 fold significant decrease in the expression pattern of PKS1 and SCD1, respectively, in B. sorokiniana treated with AgNPs. The present study provides probable understanding of molecular events underlying the antifungal role of AgNPs against B. sorokiniana.

Biosynthesized silver nanoparticles as a nanoweapon against phytopathogens: exploring their scope and potential in agriculture.

The beneficial use of silver nanoparticles (AgNPs) in agroecosystems is not fully explored with partial information available, of which most of the studies are limited to laboratory conditions and only few involve natural ecosystems. AgNPs, being the most popular metallic nanoparticles exhibiting antimicrobial property, are predominantly used for plant disease management. Owing to the ill hazards of chemically synthesized AgNPs, their biosynthesis using environment-friendly biomolecules is gaining noteworthy attention. In addition, considering the advantages of nanoformulations over biopesticides, there is no doubt that biosynthesized AgNP-based biopesticides could revolutionize the agricultural sector in the future. Though enhanced commercial use of AgNPs has generated biosafety issues in modern scenario but expecting their significant contribution towards agricultural sector, it is too early to predict the risk factor associated with their usage. To unveil the toxicity factor of AgNPs, we need to focus and understand the major interactions of AgNPs in agroecosytems. Hence, the present review highlights (i) the potential application of AgNPs in the agricultural sector particularly for plant disease management, (ii) significance of biosynthesized AgNPs using microbes and plants over their chemical synthesis, (iii) major interactions of AgNPs in agroecosystems (with soil, soil biota, and plants) with emphasis to deal with toxicity-determining factors, and (iv) identifying future research work holding promising applications of biosynthesized AgNPs in agroecosystems.

Effect of mixture of Trichoderma isolates on biochemical parameters in leaf of Macrophomina phaseolina infected brinjal.

A mixture of Trichoderma harzianum NBRI-1055 (Fx) and T. harzianum BHU-99 (Th) was evaluated for their efficiency to induce systemic resistance during three way interaction among brinjal-Trichoderma-Macrophomina phaseolina. Total phenol content (TPC), defence related enzymes Phenylalanine ammonia-lyase (PAL), Peroxidase (PO), Polyphenol oxidase (PPO) and PR proteins (PR-2 and PR-3) were recorded. Total phenolic content was recorded 12.82 times and 1.8 times higher in Trichoderma mixture treated-pathogen challenge (Fx-Th-Pth) treatment than in untreated healthy control and untreated pathogen challenged (Pth) plants respectively after 72 hr pathogen inoculation (hapi). Defence related enzymes PAL 4.54 times higher, 48hapi, PO, 3.96 times higher, 72hapi and PPO 8.1 times higher, 72hapi in Fx-Th-Pth treatment than untreated healthy control, and the PR- proteins such as PR-2, 2.15 times and PR-3, 2.16 times higher, 72hapi than untreated healthy control. The results showed that a mixture of Trichoderma (Fx+Th) performed better than single isolate.

Molecular, phenotypic and pathogenic variability in Colletotrichum isolates of subtropical region in north-eastern India, causing fruit rot of chillies.

AIM: To study the genetic variability and population structure of Colletotrichum species found to be associated with anthracnose of chilli in the north-eastern region of India. METHODS AND RESULTS: Twenty-three Colletotrichum isolates were obtained from infected chilli fruits and leaves from the chilli growing regions of north-eastern Uttar Pradesh, India, identified as Colletotrichum capsici and Colletotrichum acutatum using species-specific primers. Genetic variability among the isolates was analysed using RAPD and ISSR markers. The RAPD marker efficiently grouped the isolates at species level, while ISSR marker was effective in separating the isolates based on geographical origin. In vitro pathogenic test revealed the inability of C. acutatum isolates to infect unripe fruits, while C. capsici isolates were found to infect both ripe and unripe fruits at disease severity scale 7-9. Growth rate on different media was recorded to cross-confirm the classification of isolates, which clearly grouped the two species into distinct group on PCA plot. CONCLUSIONS: Two species, viz. C. capsici and C. acutatum, prevalent in the region were found to infect the fruits at postripening stage. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study proposes the implementation of management strategies at postripening stages of the crop to control the spread of disease, thereby leading to increment in production of chilli in the given region.

Biofabricated silver nanoparticles act as a strong fungicide against Bipolaris sorokiniana causing spot blotch disease in wheat.

The present study is focused on the extracellular synthesis of silver nanoparticles (AgNPs) using culture supernatant of an agriculturally important bacterium, Serratia sp. BHU-S4 and demonstrates its effective application for the management of spot blotch disease in wheat. The biosynthesis of AgNPs by Serratia sp. BHU-S4 (denoted as bsAgNPs) was monitored by UV-visible spectrum that showed the surface plasmon resonance (SPR) peak at 410 nm, an important characteristic of AgNPs. Furthermore, the structural, morphological, elemental, functional and thermal characterization of bsAgNPs was carried out using the X-ray diffraction (XRD), electron and atomic microscopies, energy dispersive X-ray (EDAX) spectrometer, FTIR spectroscopy and thermogravimetric analyzer (TGA), respectively. The bsAgNPs were spherical in shape with size range of ∼10 to 20 nm. The XRD and EDAX analysis confirmed successful biosynthesis and crystalline nature of AgNPs. The bsAgNPs exhibited strong antifungal activity against Bipolaris sorokiniana, the spot blotch pathogen of wheat. Interestingly, 2, 4 and 10 µg/ml concentrations of bsAgNPs accounted for complete inhibition of conidial germination, whereas in the absence of bsAgNPs, conidial germination was 100%. A detached leaf bioassay revealed prominent conidial germination on wheat leaves infected with B. sorokiniana conidial suspension alone, while the germination of conidia was totally inhibited when the leaves were treated with bsAgNPs. The results were further authenticated under green house conditions, where application of bsAgNPs significantly reduced B. sorokiniana infection in wheat plants. Histochemical staining revealed a significant role of bsAgNPs treatment in inducing lignin deposition in vascular bundles. In summary, our findings represent the efficient application of bsAgNPs in plant disease management, indicating the exciting possibilities of nanofungicide employing agriculturally important bacteria.

Trichoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani.

AIMS: To investigate the efficacy of Trichoderma harzianum NBRI-1055 (denoted as 'T-1055') in suppression of seedling blight of sunflower caused by Rhizoctonia solani Kühn and their impact on host defence responses. METHODS AND RESULTS: T-1055 was applied as seed treatment, soil application and combined application (seed treatment + soil application). Higher protection afforded by combined application of T-1055 was associated with the marked induction of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (PO) and cinnamyl alcohol dehydrogenase (CAD) activities. The activities of PAL and PPO reached maximum at 10 days after sowing (DAS), while PO and CAD levels reached maximum at 12 DAS. This was further supported by the accumulation of total phenolic content that showed an increase up to threefold at 14 DAS. In addition, HPLC analysis revealed that the contents of ferulic and p-coumaric acids increased by 6·3 and 4·6 times, respectively, at 14 DAS. Amount of gallic acid was also little more than double. Lignin deposition in sunflower root increased by 2·7, 3·4 and 3·7 times through combined application of T-1055 at 16, 18 and 20 DAS, respectively. Combined application also increased the accumulation of PR-2 and PR-3 proteins by 3·3 and 3·9 times, respectively, at 12 DAS in followed by seed treatment alone. CONCLUSIONS: The combined application of T-1055 triggered defence responses in an enhanced level in sunflower than the soil and seed alone and provided better protection against Rhizoctonia seedling blight. SIGNIFICANCE AND IMPACT OF THE STUDY: Rhizospheric fungal bioagent 'T-1055' can enhance protection in sunflower against the R. solani pathogen through augmented elicitation of host defence responses.

Quantifying global terrestrial methanol emissions using observations from the TES satellite sensor.

We employ new global space-based measurements of atmospheric methanol from the Tropospheric Emission Spectrometer (TES) with the adjoint of the GEOS-Chem chemical transport model to quantify terrestrial emissions of methanol to the atmosphere. Biogenic methanol emissions in the model are based on version 2.1 of the Model of Emissions of Gases and Aerosols from Nature (MEGANv2.1), using leaf area data from NASA's Moderate Resolution Imaging Spectroradiometer (MODIS) and GEOS-5 assimilated meteorological fields. We first carry out a pseudo observation test to validate the overall approach, and find that the TES sampling density is sufficient to accurately quantify regional- to continental-scale methanol emissions using this method. A global inversion of two years of TES data yields an optimized annual global surface flux of 122 Tg yr-1 (including biogenic, pyrogenic, and anthropogenic sources), an increase of 60 % from the a priori global flux of 76 Tg yr-1. Global terrestrial methanol emissions are thus nearly 25 % those of isoprene (~540 Tg yr-1), and are comparable to the combined emissions of all anthropogenic volatile organic compounds (~100-200 Tg yr-1). Our a posteriori terrestrial methanol source leads to a strong improvement of the simulation relative to an ensemble of airborne observations, and corroborates two other recent top-down estimates (114-120 Tg yr-1) derived using in situ and space-based measurements. Inversions testing the sensitivity of optimized fluxes to model errors in OH, dry deposition, and oceanic uptake of methanol, as well as to the assumed a priori constraint, lead to global fluxes ranging from 118 to 126 Tg yr-1. The TES data imply a relatively modest revision of model emissions over most of the tropics, but a significant upward revision in midlatitudes, particularly over Europe and North America. We interpret the inversion results in terms of specific source types using the methanol : CO correlations measured by TES, and find that biogenic emissions are overestimated relative to biomass burning and anthropogenic emissions in central Africa and southeastern China, while they are underestimated in regions such as Brazil and the US. Based on our optimized emissions, methanol accounts for > 25 % of the photochemical source of CO and HCHO over many parts of the northern extratropics during springtime, and contributes ~6 % of the global secondary source of those compounds annually.

Atmospheric peroxyacetyl nitrate (PAN): a global budget and source attribution.

Peroxyacetyl nitrate (PAN) formed in the atmospheric oxidation of non-methane volatile organic compounds (NMVOCs) is the principal tropospheric reservoir for nitrogen oxide radicals (NOx = NO + NO2). PAN enables the transport and release of NOx to the remote troposphere with major implications for the global distributions of ozone and OH, the main tropospheric oxidants. Simulation of PAN is a challenge for global models because of the dependence of PAN on vertical transport as well as complex and uncertain NMVOC sources and chemistry. Here we use an improved representation of NMVOCs in a global 3-D chemical transport model (GEOS-Chem) and show that it can simulate PAN observations from aircraft campaigns worldwide. The immediate carbonyl precursors for PAN formation include acetaldehyde (44% of the global source), methylglyoxal (30 %), acetone (7 %), and a suite of other isoprene and terpene oxidation products (19 %). A diversity of NMVOC emissions is responsible for PAN formation globally including isoprene (37 %) and alkanes (14 %). Anthropogenic sources are dominant in the extratropical Northern Hemisphere outside the growing season. Open fires appear to play little role except at high northern latitudes in spring, although results are very sensitive to plume chemistry and plume rise. Lightning NOx is the dominant contributor to the observed PAN maximum in the free troposphere over the South Atlantic.

Arthrobotrys oligospora-mediated biological control of diseases of tomato (Lycopersicon esculentum Mill.) caused by Meloidogyne incognita and Rhizoctonia solani.

AIMS: To study the biocontrol potential of nematode-trapping fungus Arthrobotrys oligospora in protecting tomato (Lycopersicon esculentum Mill.) against Meloidogyne incognita and Rhizoctonia solani under greenhouse and field conditions. METHODS AND RESULTS: Five isolates of the nematode-trapping fungus Arthrobotrys oligospora isolated from different parts of India were tested against Meloidogyne incognita and Rhizoctonia solani in tomato (Lycopersicon esculentum Mill.) plants grown under greenhouse and field conditions. Arthrobotrys oligospora-treated plants showed enhanced growth in terms of shoot and root length and biomass, chlorophyll and total phenolic content and high phenylalanine ammonia lyase activity in comparison with M. incognita- and R. solani-inoculated plants. Biochemical profiling when correlated with disease severity and intensity in A. oligospora-treated and untreated plants indicate that A. oligospora VNS-1 offered significant disease reduction in terms of number of root galls, seedling mortality, lesion length, disease index, better plant growth and fruit yield as compared to M. incognita- and R. solani-challenged plants. CONCLUSION: The result established that A. oligospora VNS-1 has the potential to provide bioprotection agents against M. incognita and R. solani. SIGNIFICANCE AND IMPACT OF THE STUDY: Arthrobotrys oligospora can be a better environment friendly option and can be incorporated in the integrated disease management module of crop protection. Application of A. oligospora not only helps in the control of nematodes but also increases plant growth and enhances nutritional value of tomato fruits. Thus, it proves to be an excellent biocontrol as well as plant growth promoting agent.

ROS-mediated apoptotic cell death in prostate cancer LNCaP cells induced by biosurfactant stabilized CdS quantum dots.

Cadmium sulfide (CdS) quantum dots (QDs) have raised great attention because of their superior optical properties and wide utilization in biological and biomedical studies. However, little is known about the cell death mechanisms of CdS QDs in human cancer cells. This study was designed to investigate the possible mechanisms of apoptosis induced by biosurfactant stabilized CdS QDs (denoted as "bsCdS QDs") in human prostate cancer LNCaP cells. It was also noteworthy that apoptosis correlated with reactive oxygen species (ROS) production, mitochondrial damage, oxidative stress and chromatin condensation in a dose- and time-dependent manner. Results also showed involvement of caspases, Bcl-2 family proteins, heat shock protein 70, and a cell-cycle checkpoint protein p53 in apoptosis induction by bsCdS QDs in LNCaP cells. Moreover, pro-apoptotic protein Bax was upregulated and the anti-apoptotic proteins, survivin and NF-κB were downregulated in bsCdS QDs exposed cells. Protection of N-acetyl cysteine (NAC) against ROS clearly suggested the implication of ROS in hyper-activation of apoptosis and cell death. It is encouraging to conclude that biologically stabilized CdS QDs bear the potential of its applications in biomedicine, such as tumor therapy specifically by inducing caspase-dependent apoptotic cell death of human prostate cancer LNCaP cells.

Lagerstroemia speciosa fruit extract modulates quorum sensing-controlled virulence factor production and biofilm formation in Pseudomonas aeruginosa.

Lagerstroemia speciosa (Lythraceae) is a south-east Asian tree more commonly known as 'Jarul'. Research on health benefits suggests that the L. speciosa plant contains phytomolecules that may have antioxidant, anti-diabetic and anti-obesity properties. However, antimicrobial activities have not been reported for this plant. The ability of L. speciosa fruit extract (LSFE) to antagonize cell-to-cell communication, expression of virulence genes and factors, and biofilm formation was evaluated in Pseudomonas aeruginosa strain PAO1. Our results suggested that LSFE caused downregulation of quorum sensing (QS)-related genes (las and rhl) and their respective signalling molecules, N-acylhomoserine lactones, without affecting the growth of P. aeruginosa PAO1. Significant inhibition of virulence factors: LasA protease, LasB elastase, and pyoverdin production, was also recorded. Application of LSFE to P. aeruginosa PAO1 biofilms increased bacterial susceptibility to tobramycin. These data suggest a possible role for quorum-quenching mechanisms unrelated to static or cidal effects, and also suggest that L. speciosa could serve as a cost-effective source in the development of new QS-based antibacterial drugs.

Tropospheric methanol observations from space: retrieval evaluation and constraints on the seasonality of biogenic emissions.

Methanol retrievals from nadir-viewing space-based sensors offer powerful new information for quantifying methanol emissions on a global scale. Here we apply an ensemble of aircraft observations over North America to evaluate new methanol measurements from the Tropospheric Emission Spectrometer (TES) on the Aura satellite, and combine the TES data with observations from the Infrared Atmospheric Sounding Interferometer (IASI) on the MetOp-A satellite to investigate the seasonality of methanol emissions from northern midlatitude ecosystems. Using the GEOS-Chem chemical transport model as an intercomparison platform, we find that the TES retrieval performs well when the degrees of freedom for signal (DOFS) are above 0.5, in which case the model:TES regressions are generally consistent with the model:aircraft comparisons. Including retrievals with DOFS below 0.5 degrades the comparisons, as these are excessively influenced by the a priori. The comparisons suggest DOFS >0.5 as a minimum threshold for interpreting retrievals of trace gases with a weak tropospheric signal. We analyze one full year of satellite observations and find that GEOS-Chem, driven with MEGANv2.1 biogenic emissions, underestimates observed methanol concentrations throughout the midlatitudes in springtime, with the timing of the seasonal peak in model emissions 1-2 months too late. We attribute this discrepancy to an underestimate of emissions from new leaves in MEGAN, and apply the satellite data to better quantify the seasonal change in methanol emissions for midlatitude ecosystems. The derived parameters (relative emission factors of 11.0, 0.26, 0.12 and 3.0 for new, growing, mature, and old leaves, respectively, plus a leaf area index activity factor of 0.5 for expanding canopies with leaf area index <1.2) provide a more realistic simulation of seasonal methanol concentrations in midlatitudes on the basis of both the IASI and TES measurements.

Dietary use and conservation concern of edible wetland plants at Indo-Burma hotspot: a case study from Northeast India.

BACKGROUND: The wetlands of the North East India fall among the global hotspots of biodiversity. However, they have received very little attention with relation to their intrinsic values to human kind; therefore their conservation is hardly addressed. These wetlands are critical for the sustenance of the tribal communities. METHODS: Field research was conducted during 2003 to 2006 in seven major wetlands of four districts of Manipur state, Northeast India (viz. Imphal-East, Imphal-West, Thoubal, and Bishnupur). A total of 224 wetland-plant-collectors were interviewed for the use and economics of species using semi-structured questionnaires and interview schedules. Imphal, Bishenpur and Thoubal markets were investigated in detail for influx and consumption pattern of these plants. The collectors were also inquired for medicinal use of wetland species. Nutritive values of 21 species were analyzed in laboratory. The vouchers were collected for all the species and deposited in the CSIR-NEIST (Formerly Regional Research Laboratory), Substation, Lamphelpat, Imphal, Manipur, India. RESULTS: We recorded 51 edible wetland species used by indigenous people for food and medicinal purposes. Thirty eight species had high medicinal values and used in the traditional system to treat over 22 diseases. At least 27 species were traded in three markets studied (i.e. Imphal, Thoubal and Bishenpur), involving an annual turnover of 113 tons of wetland edible plants and a gross revenue of Rs. 907, 770/- (US$1 = Rs. 45/-). The Imphal market alone supplies 60% of the total business. Eighty per cent of the above mentioned species are very often used by the community. The community has a general opinion that the availability of 45% species has depleted in recent times, 15 species need consideration for conservation while another 7 species deserved immediate protection measures. The nutrient analysis showed that these species contribute to the dietary balance of tribal communities. CONCLUSIONS: Considering the importance of wild wetland plants in local sustenance, it is suggested to protect their habitats, develop domestication protocols of selected species, and build programs for the long-term management of wetland areas by involving local people. Some medicinal plants may also be used to develop into modern medicines.

Genetically engineered bacteria: an emerging tool for environmental remediation and future research perspectives.

This minireview explores the environmental bioremediation mediated by genetically engineered (GE) bacteria and it also highlights the limitations and challenges associated with the release of engineered bacteria in field conditions. Application of GE bacteria based remediation of various heavy metal pollutants is in the forefront due to eco-friendly and lesser health hazards compared to physico-chemical based strategies, which are less eco-friendly and hazardous to human health. A combination of microbiological and ecological knowledge, biochemical mechanisms and field engineering designs would be an essential element for successful in situ bioremediation of heavy metal contaminated sites using engineered bacteria. Critical research questions pertaining to the development and implementation of GE bacteria for enhanced bioremediation have been identified and poised for possible future research. Genetic engineering of indigenous microflora, well adapted to local environmental conditions, may offer more efficient bioremediation of contaminated sites and making the bioremediation more viable and eco-friendly technology. However, many challenges are to be addressed concerning the release of genetically engineered bacteria in field conditions. There are possible risks associated with the use of GE bacteria in field condition, with particular emphasis on ways in which molecular genetics could contribute to the risk mitigation. Both environmental as well as public health concerns need to be addressed by the molecular biologists. Although bioremediation of heavy metals by using the genetically engineered bacteria has been extensively reviewed in the past also, but the bio-safety assessment and factors of genetic pollution have been never the less ignored.

Anti-tuberculosis activity of selected medicinal plants against multi-drug resistant Mycobacterium tuberculosis isolates.

BACKGROUND & OBJECTIVES: Emergence of multi-drug resistant (MDR) and extensively-drug resistant (XDR) strains of Mycobacterium tuberculosis has further complicated the problem of tuberculosis (TB) control. Medicinal plants offer a hope for developing alternate medicines for the treatment of TB. The present study was done to evaluate in vitro anti-tubercular activity of five medicinal plants viz., Acalypha indica, Adhatoda vasica, Allium cepa, Allium sativum and Aloe vera. METHODS: Aqueous extracts of leaves of A. indica, A. vasica, bulbs of A. cepa, cloves of A. sativum and pure gel of A. vera leaves, were tested in vitro for their activity against two MDR isolates (DKU-156 and JAL-1236), reference susceptible strain M. tuberculosis H37Rv as well as rapid grower mycobacterial pathogen M. fortuitum (TMC-1529) using Lowenstein Jensen (L-J) medium and colorimetric BacT/ ALERT 3D system. Activity in L-J medium was evaluated by percentage inhibition which was calculated by mean reduction in number of colonies on extract containing as compared to extract free controls. RESULTS: Extracts of all the five plants A. indica, A. vasica, A. cepa, A. sativum and A. vera exhibited anti-tuberculosis activity in L-J medium, the proportion of inhibition of these plants extract in respect mentioned above is 95, 32, 37, 72, 32 per cent, respectively for MDR isolate DKU-156 and 68, 86, 79, 72, 85 per cent, respectively for another MDR isolate JAL-1236, while for sensitive M. tuberculosis H37Rv, inhibition was found to be 68, 70, 35, 63 and 41 per cent, at 4 per cent v/v concentration in L-J medium. There was no inhibition against rapid grower M. fortuitum (TMC-1529). In BacT/ALERT also, extracts of these plants showed significant inhibition against M. tuberculosis. INTERPRETATION & CONCLUSION: Our findings showed that all these plants exhibited activity against MDR isolates of M. tuberculosis. While the anti-TB activity of A. vera, A. vasica and A. sativum against MDR isolates confirm earlier results, activity of the extracts of A. indica and A. cepa is reported for the first time. Further studies aimed at isolation and identification of active substances from the extracts which exhibited promising activities, need to be carried out.

Oxidative DNA damage protective activity, antioxidant and anti-quorum sensing potentials of Moringa oleifera.

The aqueous extract of leaf (LE), fruit (FE) and seed (SE) of Moringa oleifera was assessed to examine the ability to inhibit the oxidative DNA damage, antioxidant and anti-quorum sensing (QS) potentials. It was found that these extracts could significantly inhibit the OH-dependent damage of pUC18 plasmid DNA and also inhibit synergistically with trolox, with an activity sequence of LE > FE > SE. HPLC and MS/MS analysis was carried out, which showed the presence of gallic acid, chlorogenic acid, ellagic acid, ferulic acid, kaempferol, quercetin and vanillin. The LE was with comparatively higher total phenolics content (105.04 mg gallic acid equivalents (GAE)/g), total flavonoids content (31.28 mg quercetin equivalents (QE)/g), and ascorbic acid content (106.95 mg/100 g) and showed better antioxidant activity (85.77%), anti-radical power (74.3), reducing power (1.1 ascorbic acid equivalents (ASE)/ml), inhibition of lipid peroxidation, protein oxidation, OH-induced deoxyribose degradation, and scavenging power of superoxide anion and nitric oxide radicals than did the FE, SE and standard alpha-tocopherol. Eventually, LE and FE were found to inhibit violacein production, a QS-regulated behavior in Chromobacterium violaceum 12472.