Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

Background  

Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data.     

Modeling the water budget in a deep caldera lake and its hydrologic assessment: Lake Ikeda, Japan

The hydrologic assessment of a lake water budget can be helpful in achieving proper water management and sustainable water use. A model to analyze a lake water budget was developed and verified for Lake Ikeda, Japan. Lake evaporation was estimated by numerical analyses of lake water temperature and the lake energy budget. Inflow from the lake catchment area and leakage from the lake bottom were estimated based on the tank model and Darcy's law, and the model parameters were optimized by the shuffled complex evolution method. The estimated monthly lake evaporation rate is consistent with the evaporation rate estimated by the energy budget Bowen ratio method based on in situ data from 2004 to 2005. Moreover, the calculated time series of daily lake levels agrees well with those of measured lake levels during 1983 to 1999. Thus, the model is useful for evaluating the lake water budget. Numerical analysis reveals seasonal and annual variation characteristics in the water budget components. Precipitation, inflow from the catchment area, and river water supply are generally high during the rainy season from June to July with substantial annual variation. Lake evaporation is greatest in October and least in April, but the annual variation is relatively small. Agricultural water use is relatively high from April to September. There are no marked seasonal changes in leakage and drinking water use. The lake level is generally highest in September and lowest in March, which is characterized by seasonal changes in water budget components. The model was also applied to 17-year simulations under hypothetical hydrologic conditions to examine the effect of water use and agricultural water management on the lake level. Results indicate that river water supply, provided under the agricultural water management system, effectively compensates for the decrease in lake water resulting from agricultural water use.     

Integrated management of irrigation water and fertilizers for wheat crop using field experiments and simulation modeling

The reported study aimed at developing an integrated management strategy for irrigation water and fertilizers in case of wheat crop in a sub-tropical sub-humid region. Field experiments were conducted on wheat crop (cultivar Sonalika) during the years 2002–2003, 2003–2004 and 2004–2005. Each experiment included four fertilizer treatments and three irrigation treatments during the wheat growth period. During the experiment, the irrigation treatments considered were I₁ = 10% maximum allowable depletion (MAD) of available soil water (ASW); I₂ = 40% MAD of ASW; I₃ = 60% MAD of ASW. The fertilizer treatments considered in the experiments were F₁ = control treatment with N:P₂O₅:K₂O as 0:0:0 kg ha⁻¹, F₂ = fertilizer application of N:P₂O₅:K₂O as 80:40:40 kg ha⁻¹; F₃ = fertilizer application of N:P₂O₅:K₂O as 120:60:60 kg ha⁻¹ and F₄ = fertilizer application of N:P₂O₅:K₂O as 160:80:80 kg ha⁻¹. In this study CERES-wheat crop growth model of the DSSAT v4.0 was used to simulate the growth, development and yield of wheat crop using soil, daily weather and management inputs, to aid farmers and decision makers in developing strategies for effective management of inputs. The results of the investigation revealed that magnitudes of grain yield, straw yield and maximum LAI of wheat crop were higher in low volume high frequency irrigation (I₁) than the high volume low frequency irrigation (I₃). The grain yield, straw yield and maximum LAI increased with increase in fertilization rate for the wheat crop. The results also revealed that increase in level of fertilization increased water use efficiency (WUE) considerably. However, WUE of the I₂ irrigation schedule was comparatively higher than the I₁ and I₃ irrigation schedules due to higher grain yield per unit use of water. Therefore, irrigation schedule with 40% maximum allowable depletion of available soil water (I₂) could safely be maintained during the non-critical stages to save water without sacrificing the crop yield. Increase in level of fertilization increases the WUE but it will cause environmental problem beyond certain limit. The calibrated CERES-wheat model could predict the grain yield, straw yield and maximum LAI of wheat crop with considerable accuracy and therefore can be recommended for decision-making in similar regions.     

Integrated effect of transplanting date, cultivar and irrigation on yield, water saving and water productivity of rice (Oryza sativa L.) in Indian Punjab: Field and simulation study

Individual effect of different field scale management interventions for water saving in rice viz. changing date of transplanting, cultivar and irrigation schedule on yield, water saving and water productivity is well documented in the literature. However, little is known about their integrated effect. To study that, field experimentation and modeling approach was used. Field experiments were conducted for 2 years (2006 and 2007) at Punjab Agricultural University Farm, Ludhiana on a deep alluvial loamy sand Typic Ustipsamment soils developed under hyper-thermic regime. Treatments included three dates of transplanting (25 May, 10 June and 25 June), two cultivars (PR 118 inbred and RH 257 hybrid) and two irrigation schedules (2-days drainage period and at soil water suction of 16 kPa). The model used was CropSyst, which has already been calibrated for growth (periodic biomass and LAI) of rice and soil water content in two independent experiments. The main findings of the field and simulation studies conducted are compared to any individual, integrated management of transplanting date, cultivar and irrigation, sustained yield (6.3-7.5 t ha⁻¹) and saved substantial amount of water in rice. For example, with two management interventions, i.e. shifting of transplanting date to lower evaporative demand (from 5 May to 25 June) concomitant with growing of short duration hybrid variety (90 days from transplanting to harvest), the total real water saving (wet saving) through reduction in evapotranspiration (ET) was 140 mm, which was almost double than managing the single, i.e. 66 mm by shifting transplanting or 71 mm by growing short duration hybrid variety. Shifting the transplanting date saved water through reduction in soil water evaporation component while growing of short duration variety through reduction in both evaporation and transpiration components of water balance. Managing irrigation water schedule based on soil water suction of 16 kPa at 15-20 cm soil depth, compared to 2-day drainage, did not save water in real (wet saving), however, it resulted into apparent water saving (dry saving). The real crop water productivity (marketable yield/ET) was more by 17% in 25th June transplanted rice than 25th May, 23% in short duration variety than long and 2% in irrigation treatment of 16 kPa soil water suction than 2-days drainage. The corresponding values for the apparent crop water productivity (marketable yield/irrigation water applied) were 16, 20 and 50%, respectively. Pooled experimental data of 2 years showed that with managing irrigation scheduling based on soil water suction of 16 kPa at 15-20 cm soil depth, though 700 mm irrigation water was saved but the associated yield was reduced by 277 kg ha⁻¹.     

Mineral content of three olive cultivars irrigated with treated industrial wastewater

The reuse of saline treated industrial wastewater generated by textile firms mixed with municipal domestic effluent for irrigation was used to asses its effect on the mineral content of three olive (Olea europaea L.) cultivars under greenhouse and field conditions during two complete vegetative cycles. Chemical analysis of the treated wastewater indicated that the element concentrations fall within the permissible range of irrigation water used for plants. However, little impermissible accumulation of Na and Mg higher than the recommended maximum concentration was observed. Irrigation water with six electrical conductivities (EC = 0.78, 1.0, 2.0, 3.0, 4.0 and 5.0 dS m⁻¹ in treatments T₀, T₁, T₂, T₃, T₄, T₅, respectively) were compared in the greenhouse experiment. The olive trees in the field experiment were trickle irrigated with potable water and treated wastewater (average EC = 4.2 dS m⁻¹). The results of the greenhouse experiment showed that leaf N, Cu, Mn, Fe, Pb, and Na contents increased with increasing salinity of the treated wastewater. This increase was accompanied with a decrease in K and Mg contents. Leaf Ca and Cl concentrations were not considerably affected. Ion analysis in roots indicated that the contents of P, Na, Cl, Mn, and Pb increased while K decreased as treated wastewater salinity increased. Consequently, in most cases T₄ and T₅ gave a highly significant increase or decrease in accumulation of the previously mentioned minerals. A considerable variation in the studied cultivars was noticed. ‘Nabali’ was considered the most tolerant cultivar for the high salinity levels of the treated wastewater; its transporting selectivity of Na from root to leaf was higher and more Na was retained in the roots. Tissue analysis of leaves indicated that the element concentrations were within the adequate levels except those of Fe in ‘Nabali’ and ‘Manzanillo’, Na in ‘Improved Nabali’ and Cu in ‘Nabali’ and ‘Manzanillo’. In view of these findings, the negligible accumulation of minerals in leaves and roots indicated that this kind of textile effluent can be used as a valid alternative for irrigation of olive orchards with continuous monitoring of mineral levels.     

Purification and characterization of a 630 kDa bacterial killing metalloprotease (KilC) isolated from plaice Pleuronectes platessa (L.), epidermal mucus

Antibacterial chemicals in the mucus of fish such as lysozyme, lectins, peptides and proteases provide an efficient first line of defence against pathogens. This study shows that there are at least three antibacterial proteins in plaice skin mucus in addition to lysozyme. One of these proteins is responsible for approximately 74% of the antibacterial activity and is a 630 kDa protease complex designated KilC (bacterial killing metalloprotease C). Purified KilC kills the bacteria Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa efficiently. The protease activity of KilC is dependent upon the divalent cation Mg(2+) and shows pH dual optima of 5.0 and 8.0. The enzyme has a temperature optimum of 25 degrees C and is made up of at least five different sized peptides. Studies with protease inhibitors show that the catalytic site of KilC may be cysteine- or serine protease-like. KilC may kill bacterial cells by acting directly upon the bacteria or by producing low molecular weight bioactive compounds such as peptides.     

Enhancing the mineral and vitamin content of wheat and maize through plant breeding

More than half of the world's population suffers micronutrient undernourishment. The main sources of vitamins and minerals (iron, zinc, and vitamin A) for low-income rural and urban populations are staple foods of plant origin that often contain low levels or low bioavailability of these micronutrients. Biofortification aims to develop micronutrient-enhanced crop varieties through conventional plant breeding. HarvestPlus, the CGIAR's biofortification initiative, seeks to breed and disseminate crop varieties with enhanced micronutrient content that can improve the nutrition of the “hard to reach” (by fortification or supplementation programmes) rural and urban poor in targeted countries/regions. In attempting to enhance micronutrient levels in maize and wheat through conventional plant breeding, it is important to identify genetic resources with high levels of the targeted micronutrients, to consider the heritability of the targeted traits, to explore the availability of high throughput screening tools and to gain a better understanding of genotype by environment interactions. Biofortified maize and wheat varieties must have the trait combinations which encourage adoption such as high yield potential, disease resistance, and consumer acceptability. When defining breeding strategies and targeting micronutrient levels, researchers need to consider the desired micronutrient increases, food intake and retention and bioavailability as they relate to food processing, anti-nutritional factors and promoters. Finally, ex ante studies are required to quantify the burden of micronutrient deficiency and the potential of biofortification to achieve a significant improvement in human micronutrient status in the deficient target population in order to determine whether a biofortification program is cost-effective.     

Effects of transition season management on soil N dynamics and system N balances in rice–wheat rotations of Nepal

In the low-input rice–wheat production systems of Nepal, the N nutrition of both crops is largely based on the supply from soil pools. Declining yield trends call for management interventions aiming at the avoidance of native soil N losses. A field study was conducted at two sites in the lowland and the upper mid-hills of Nepal with contrasting temperature regimes and durations of the dry-to-wet season transition period between the harvest of wheat and the transplanting of lowland rice. Technical options included the return of the straw of the preceding wheat crop, the cultivation of short-cycled crops during the transition season, and combinations of both. Dynamics of soil N_min, nitrate leaching, nitrous oxide emissions, and crop N uptake were studied throughout the year between 2004 and 2005 and partial N balances of the cropping systems were established. In the traditional system (bare fallow between wheat and rice) a large accumulation of soil nitrate N and its subsequent disappearance upon soil saturation occurred during the transition season. This nitrate loss was associated with nitrate leaching (6.3 and 12.8 kg ha⁻¹ at the low and high altitude sites, respectively) and peaks of nitrous oxide emissions (120 and 480 mg m⁻² h⁻¹ at the low and high altitude sites, respectively). Incorporation of wheat straw at 3 Mg ha⁻¹ and/or cultivation of a nitrate catch crop during the transition season significantly reduced the build up of soil nitrate and subsequent N losses at the low altitude site. At the high altitude site, cumulative grain yields increased from 2.35 Mg ha⁻¹ with bare fallow during the transition season to 3.44 Mg ha⁻¹ when wheat straw was incorporated. At the low altitude site, the cumulative yield significantly increased from 2.85 Mg ha⁻¹ (bare fallow) to between 3.63 and 6.63 Mg ha⁻¹, depending on the transition season option applied. Irrespective of the site and the land use option applied during the transition season, systems N balances remained largely negative, ranging from −37 to −84 kg N ha⁻¹. We conclude that despite reduced N losses and increased grain yields the proposed options need to be complemented with additional N inputs to sustain long-term productivity.     

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.)

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.