In vitro production of thiophenes using hairy root cultures of Tagetes erecta (L.)

Marigold (Tagetes spp.) is a source of thiophenes, which are a group of heterocyclic sulfurous compounds  possessing strong biocidal activity, thus making Tagetes plants very useful as natural source of agents for controlling pathogens such as nematodes, insects, fungi and bacteria. Hairy root cultures of Tagetes erecta L. were developed using Agrobacterium rhizogenes mediated transformation. The hairy root clones showed great variations in growth pattern and total thiophene content (0.31 to 0.96 mg/gfw). Four types of thiophenes that accumulated in root cultures of Tagetes were butenenylbithiophene (BBT),  hydroxybutenenylbithiophene (BBTOH), acetoxybutenenylbithiophene (BBTOAc) and α-terthienyl (α-T). Total thiophene contents in these isolated rhizoclones were found tobe four fold higher than that of wild type hairy root cultures. The developed method of producing hairy cultures of T. erecta can be used for producing thiophenes at large scale.Key words: Agrobacterium rhizogenes, marigold, rhizoclones, biocidal

In vitro Direct Regeneration and Agrobacterium Tumefaciens mediated in planta Transformation of Ocimum sanctum L.

An in vitro regeneration system for propagation has been successfully developed for a valuable medicinal and aromatic plant ‘Ocimum sanctum L’. In the present study, petiole explants, from in-vitro grown cultures of O. sanctum, was used for direct regeneration. The developed protocol employed 98% of regeneration frequency in addition to 9.6 shoots per explant when cultured on Murashige and Skoog (MS) medium fortified with 3 mg/L benzylamino purine (BAP) and 1 mg/L Naphthalene acetic acid (NAA). Furthermore, Agrobacterium tumefaciens Mediated genetic Transformation (ATMT) protocol (transient and stable) was also developed using LBA4404 strain harboring pBI121 with uid-A and neomycin phosphotransferase genes. The regenerated transformants were shifted on MS with kanamycin (50 mg/L) and afterwards placed on the half-strength MS medium. The validation was done through polymerase chain reaction (PCR) with neomycin phosphotransferase-II (npt-II) & β-glucoronidase (uid-A) gene primers. The maximum stable transformation frequency of 70% ± 0.35 was achieved. Hence, it is apparent that the established protocols i.e. in vitro direct regeneration and ATMT are appropriate for integrating novel enzymes/genes through high throughput techniques such as gene tagging, and targeted gene replacement to modulate the primary as well as secondary metabolic flux towards desired agronomic product or trait in planta

Smart city-ranking of major Australian cities to achieve a smarter future

© 2020 by the authors. A Smart City is a solution to the problems caused by increasing urbanization. Australia has demonstrated a strong determination for the development of Smart Cities. However, the country has experienced uneven growth in its urban development. The purpose of this study is to compare and identify the smartness of major Australian cities to the level of development in multi-dimensions. Eventually, the research introduces the openings to make cities smarter by identifying the focused priority areas. To ensure comprehensive coverage of all aspects of the smart city's performance, 90 indicators were selected to represent 26 factors and six components. The results of the assessment endorse the impacts of recent government actions taken in different urban areas towards building smarter cities. The research has pointed out the areas of deficiencies for underperforming major cities in Australia. Following the results, appropriate recommendations for Australian cities are provided to improve the city's smartness

Identification of Major Inefficient Water Consumption Areas Considering Water Consumption, Efficiencies, and Footprints in Australia

Due to population growth, climatic change, and growing water usage, water scarcity is expected to be a more prevalent issue at the global level. The situation in Australia is even more serious because it is the driest continent and is characterized by larger water footprints in the domestic, agriculture and industrial sectors. Because the largest consumption of freshwater resources is in the agricultural sector (59%), this research undertakes a detailed investigation of the water footprints of agricultural practices in Australia. The analysis of the four highest water footprint crops in Australia revealed that the suitability of various crops is connected to the region and the irrigation efficiencies. A desirable crop in one region may be unsuitable in another. The investigation is further extended to analyze the overall virtual water trade of Australia. Australia’s annual virtual water trade balance is adversely biased towards exporting a substantial quantity of water, amounting to 35 km3, per trade data of 2014. It is evident that there is significant potential to reduce water consumption and footprints, and increase the water usage efficiencies, in all sectors. Based on the investigations conducted, it is recommended that the water footprints at each state level be considered at the strategic level. Further detailed analyses are required to reduce the export of a substantial quantity of virtual water considering local demands, export requirements, and production capabilities of regions

Qualitative analysis of essential oil of Rosmarinus officinalis L. cultivated in Uttaranchal Hills, India

Gas chromatography and gas chromatography and mass spectroscopy analysis of essentialoil hydrodistilled from fresh aerial part of rosemary (Rosmarinus officinalis) cultivated inUttaranchal, India, resolved into 33 compounds representing 96.29% of the essential oilaccording to their chromatographic retention indices and mass spectra.  The major compoundsof the oil were, camphor (26.40%), 1, 8-cineole (23.40%), α-pinene (9.94%), camphene (5.83%),myrcene (4.86%), bornyl acetate (3.97%), verbenone (3.32%), limonene (3.08%), borneol(2.05%) and α-terpineol (2.68%). &nbsp

Design and implementation of Adaptive Neuro-Fuzzy Inference system for the control of an uncertain Ball on Beam Apparatus

Controlling an uncertain mechatronic system is challenging and crucial for its automation. In this regard, several control-strategies are developed to handle such systems. However, these control-strategies are complex to design, and require in-depth knowledge of the system and its dynamics. In this study, we are testing the performance of a rather simple control-strategy (Adaptive Neuro-Fuzzy Inference System) using an uncertain Ball and Beam System. The custom-designed apparatus utilizes image processing technique to acquire the position of the ball on the beam. Then, desired position is achieved by controlling the beam angle using Adaptive Neuro-Fuzzy and PID control. We are showing that adaptive neuro-fuzzy control can effectively handle the system uncertainties, which traditional controllers (i.e., PID) cannot handle

Prediction of the amount of sediment Deposition in Tarbela Reservoir using machine learning approaches

Tarbela is the largest earth-filled dam in Pakistan, used for both irrigation and power production. Tarbela has already lost around 41.2% of its water storage capacity through 2019, and WAPDA predicts that it will continue to lose storage capacity. If this issue is ignored for an extended period of time, which is not far away, a huge disaster will occur. Sedimentation is one of the significant elements that impact the Tarbela reservoir’s storage capacity. Therefore, it is crucial to accurately predict the sedimentation inside the Tarbela reservoir. In this paper, an Artificial Neural Network (ANN) architecture and multivariate regression technique are proposed to validate and predict the amount of sediment deposition inside the Tarbela reservoir. Four input parameters on yearly basis including rainfall (Ra), water inflow (Iw), minimum water reservoir level (Lr), and storage capacity of the reservoir (Cr) are used to evaluate the proposed machine learning models. Multivariate regression analysis is performed to undertake a parametric study for various combinations of influencing parameters. It was concluded that the proposed neural network model estimated the amount of sediment deposited inside the Tarbela reservoir more accurately as compared to the multivariate regression model because the maximum error in the case of the proposed neural network model was observed to be 4.01% whereas in the case of the multivariate regression model was observed to be 60.7%. Then, the validated neural network model was used for the prediction of the amount of sediment deposition inside the Tarbela reservoir for the next 20 years based on the time series univariate forecasting model ETS forecasted values of Ra, Iw, Lr, and Cr. It was also observed that the storage capacity of the Tarbela reservoir is the most influencing parameter in predicting the amount of sediment

Development of a Hydrodynamic-Based Flood-Risk Management Tool for Assessing Redistribution of Expected Annual Damages in a Floodplain

Despite spending ample resources and procedural development in flood management, flood losses are still increasing worldwide. The losses caused by floods and costs incurred on management are two components of expected annual damages (EAD) due to floods. This study introduces a generalized approach for risk-based design where a range of probable floods are considered before and after a flood mitigation measure is implemented. The proposed approach is customized from the ISO Guide 31000 along with additional advantages of flood risk visualization. A Geographic Information System (GIS)-based design of a flood-protection dike is performed to exhibit the risk redistribution. The Chenab River is selected for the existing dike system. Detailed hazard behaviour and societal vulnerability are modelled and visualized for a range of all probable floods before and after the implementation of flood-protection dikes. EAD maps demonstrate the redistribution of induced and residual risks. It can be concluded that GIS-based EAD maps not only facilitate cost-effective solutions but also provide an accurate estimate of residual risks after the mitigation measures are applied. EAD maps also indicate the high-risk areas to facilitate designing secondary measures

Root tip-dependent, active riboflavin secretion by Hyoscyamus albus hairy roots under iron deficiency

Hyoscyamus albus hairy roots with/without an exogenous gene (11 clones) were established by inoculation of Agrobacterium rhizogenes. All clones cultured under iron deficient condition secreted riboflavin from root tips into the culture medium and the productivity depended on the number and size of root tips among the clones, although the addition of sucrose was essential for riboflavin production. A decline of pH was observed before riboflavin production and root development using either a root tip or propagated roots: propagated roots were employed for further work due to their lesser variation. Additions of proton-pump inhibitors, N,N’-dicyclohexylcarbodiimide (DCCD) at 100 and 10 μM and erythrosine B at 100 μM, suppressed the pH decline at 100 and 10 μM accompanied by inhibition of riboflavin secretion and root growth; at 10 μM of erythrosine B, pH decline occurred with a moderate delay, but both growth and riboflavin efflux were inhibited. Neither inhibition of the pH decline nor riboflavin production was observed at 1 μM. To examine the necessity of acidification and riboflavin secretion by the roots themselves, artificial pH reduction of culture medium with organic acids and the addition of exogenous riboflavin with/without pH reduction were performed. When hairy roots were cultured in iron-deficient medium acidified with citric acid (pH 4.0) or malic acid (pH 3.7), pH increased rapidly to around 5 overnight, following which riboflavin production and root growth occurred. Addition of riboflavin did not affect riboflavin secretion by the roots, but acidification with citric acid (pH 4.0) helped achiever greater riboflavin production and earlier pH elevation. These results indicate that riboflavin efflux does not directly connected to active pH reduction, and more significantly active riboflavin secretion occurs by internal requirement in H. albus hairy roots under iron deficiency

HCHL expression in hairy roots of Beta vulgaris yields a high accumulation of p-hydroxybenzoic acid (pHBA) glucose ester, and linkage of pHBA into cell walls.

As part of a study to explore the potential for new or modified bio-product formation, Beta vulgaris (sugar beet) has been genetically modified to express in root-organ culture a bacterial gene of phenylpropanoid catabolism. The HCHL gene, encoding p-hydroxycinnamoyl-CoA hydratase/lyase, was introduced into B. vulgaris under the control of a CaMV 35S promoter, using Agrobacterium rhizogenes LBA 9402. Hairy root clones expressing the HCHL gene, together with non-expressing clones, were analysed and revealed that one expression-positive clone accumulated the glucose ester of p-hydroxybenzoic acid (pHBA) at about 14% on a dry weight basis. This is the best yield achieved in plant systems so far. Determination of cell-wall components liberated by alkaline hydrolysis confirmed that the ratio of pHBA to ferulic acid was considerably higher in the HCHL-expressing clones, whereas only ferulic acid was detected in a non-expressing clone. The change in cell-wall components also resulted in a decrease in tensile strength in the HCHL-expressing clones