Effects of seasonal variations and pond age on trace elements and their correlations with plankton productivity in commercial freshwater crayfish (cherax cainii austin, 2002) earthen ponds

Seasonal variations can affect the concentration of trace elements, and the change in their concentrations can affect the natural productivity of freshwater aquaculture ponds. Hence, we studied the seasonal variations of the 12 pre‐selected trace elements (Co, Cu, Fe, Mn, Zn, Se, Ca, Mg, P, S, Al, Si) and their relationships with primary and secondary productivity in two aged ponds, stocked with three different life stages of marron (Cherax cainii), for a period of 1 year. Trace element analysis was performed by using (Agilent, ICP‐OES). Except Co and Se, all trace elements, and primary and secondary productivity were influenced by seasonal variation. The pond age significantly influenced the concentrations of some trace elements. On a seasonal basis, trace elements were positively correlated with the plankton abundance, species diversity, and wet and dry plankton weights. Seasonal variations and pond age affected the dissolved concentrations of trace elements and plankton productivity.     

Universal endogenous gene controls for bisulphite conversion in analysis of plant DNA methylation

Background

Protein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, plant materials usually have to be ground and homogenized to physically break the robust cell wall, but this step is laborious and time-consuming when a large number of samples are handled at once.

Results

We developed a chemical method for lysing Arabidopsis cells without grinding. In this method, plants are boiled for just 10 minutes in a solution containing a Ca²⁺ chelator and detergent. Cell extracts prepared by this method were suitable for SDS-PAGE and immunoblot analysis. This method was also applicable to genomic DNA extraction for PCR analysis. Our method was applied to many other plant species, and worked well for some of them.

Conclusions

Our method is rapid and economical, and allows many samples to be prepared simultaneously for protein analysis. Our method is useful not only for Arabidopsis research but also research on certain other species.     

High Resolution Melt (HRM) analysis is an efficient tool to genotype EMS mutants in complex crop genomes

Background

Targeted Induced Loci Lesions IN Genomes (TILLING) is increasingly being used to generate and identify mutations in target genes of crop genomes. TILLING populations of several thousand lines have been generated in a number of crop species including Brassica rapa. Genetic analysis of mutants identified by TILLING requires an efficient, high-throughput and cost effective genotyping method to track the mutations through numerous generations. High resolution melt (HRM) analysis has been used in a number of systems to identify single nucleotide polymorphisms (SNPs) and insertion/deletions (IN/DELs) enabling the genotyping of different types of samples. HRM is ideally suited to high-throughput genotyping of multiple TILLING mutants in complex crop genomes. To date it has been used to identify mutants and genotype single mutations. The aim of this study was to determine if HRM can facilitate downstream analysis of multiple mutant lines identified by TILLING in order to characterise allelic series of EMS induced mutations in target genes across a number of generations in complex crop genomes.

Results

We demonstrate that HRM can be used to genotype allelic series of mutations in two genes, BraA.CAX1a and BraA.MET1.a in Brassica rapa. We analysed 12 mutations in BraA.CAX1.a and five in BraA.MET1.a over two generations including a back-cross to the wild-type. Using a commercially available HRM kit and the Lightscanner? system we were able to detect mutations in heterozygous and homozygous states for both genes.

Conclusions

Using HRM genotyping on TILLING derived mutants, it is possible to generate an allelic series of mutations within multiple target genes rapidly. Lines suitable for phenotypic analysis can be isolated approximately 8-9 months (3 generations) from receiving M₃ seed of Brassica rapa from the RevGenUK TILLING service.     

Lactic Acid Fermentation of β-Carotene Rich Sweet Potato (<Emphasis Type="Italic">Ipomoea batatas</Emphasis> L.) into Lacto-juice

Lacto-juices processed by lactic acid fermentation bring about a change in the beverage assortment for their high nutritive value, vitamins and minerals which are beneficial to human health when consumed. Sweet potato roots (non-boiled/ fully-boiled) were fermented with Lactobacillus plantarum MTCC 1407 at 28 ± 2°C for 48 h to make lacto- juice. During fermentation both analytical [pH, titratable acidity, lactic acid, starch, total sugar, reducing sugar (g/kg roots), total phenol and β-carotene (mg/kg roots)] and sensory (texture, taste, aroma, flavour and after taste) analyses of sweet potato lacto-juice were evaluated. The fermented juice was subjected to panelist evaluation for acceptability. There were no significant variations in biochemical constituents (pH, 2.2–3.3; lactic acid, 1.19–1.27 g/kg root; titratable acidity, 1.23–1.46 g/kg root, etc.) of lacto-juices prepared from non-boiled and fully-boiled sweet potato roots except β-carotene concentration [130 ± 7.5 mg/kg (fully-boiled roots) and 165 ± 8.1 mg/kg (non-boiled roots)]. The panelist evaluation scores ranged from 3–4.8 (in a hedonic scale of 1–5) from moderate liking to very much liking of sweet potato lacto-juice. Principal component analyses reduced the eight original analytical variables to three independent components (factors), which accounted for 99.9% of the total variations. Similarly, five original sensory variables were reduced to two independent components, which accounted for 83.1% of the total variations.     

Equilibrium and Kinetic Studies on Biosorption of Heavy Metals by Leaf Powder of Paper Mulberry (Broussonetia papyrifera)

Paper mulberry (Broussonetia papyrifera) leaf powder was used to remove heavy metal ions from aqueous solutions. The specific uptakes of Cu (II), Pb (II), and Cd (II) by the leaf powder were 43.40?±?0.2, 43.9?±?0.5, and 30.65?±?0.9 mg g^?1, respectively, when 500 mg L^?1 of the metal solutions were used. The data fitted well to the Langmuir isotherm. The process followed the pseudo-second-order kinetic equation and intraparticle diffusion played an important role in the adsorption process. On the basis of the calculated thermodynamic parameters such as standard enthalpy (??H°), entropy (??S°) and free energy change (??G°), it was inferred that the sorption process was endothermic and spontaneous in nature. The surface properties of the leaf powder (revealed by scanning electron microscopic observations) were suitable for the metal adsorption process. Energy dispersive X-ray fluorescence analysis confirmed the sequestration of the metal ions by the leaf powder. Fourier transform infrared spectroscopy implicated that different functional groups on the leaf powder were involved in the metal adsorption process. The results obtained from this study implicated that the B. papyrifera leaf powder was a good choice as a metal adsorbent. This abundantly available natural and eco-friendly biosorbent could be effectively used to develop a technology in the future.     

Enhanced soybean (Glycine max L.) plant growth and nodulation by Bradyrhizobium japonicum-SB1 in presence of Bacillus thuringiensis-KR1

Abstract

Nodulation and subsequent nitrogen fixation are important factors that determine the productivity of soybean (Glycine max L.). The beneficial effects of nodulation can be enhanced when rhizobial inoculation is combined with plant-growth-promoting bacteria (PGPB). The PGPB strain Bacillus thuringiensis-KR1, originally isolated from the nodules of Kudzu vine (Pueraria thunbergiana), was found to promote growth of soybean plants (variety VL Soya 2) under Jensen's tube and growth pouch conditions, when co-inoculated with Bradyrhizobium japonicum-SB1. Co-inoculation with Bacillus thuringiensis-KR1 (at a cell density of 10 cfu) provided the highest and most consistent increase in nodule number, shoot weight, root weight, root volume, and total biomass, over rhizobial inoculation and control, under both conditions. The results demonstrate the potential benefits of using nonrhizobial nodule occupants of wild legumes for the co-inoculation of soybean, with Bradyrhizobium japonicum-SB1, in order to achieve plant-growth promotion and increased nodulation.     

Antibiofilm potential of a tropical marine Bacillus licheniformis isolate: role in disruption of aquaculture associated biofilms

Microbial biofilms are important in aquaculture industries as they resist antibiotic treatments. In this study, we have investigated the antibiofilm potential of a tropical marine culture Bacillus licheniformis D1 (containing an antimicrobial protein BLDZ1) against two aquaculture associated pathogens namely, Vibrio harveyi and Pseudomonas aeruginosa. Both the test cultures formed biofilms on polystyrene and glass surfaces. The cell free supernatant (CFS) of B. licheniformis inhibited V. harveyi and P. aeruginosa biofilms on polystyrene surfaces up to around 80% and 78% respectively. In addition, the CFS disrupted pre‐formed biofilms of test cultures by about 73%. Fluorescence and scanning electron microscope analysis confirmed the antibiofilm potential of the CFS. The cell free supernatant displayed antiadhesive activity that inhibited the initial attachment of the bacteria during the process of biofilm formation. In addition, the CFS exhibited antimicrobial activity and mediated cell death via cytoplasmic membrane disruption.     

Effects of an oil-based substrate (The Water Cleanser™) and bacterial additives on nitrogen and phosphorous dynamics in freshwater crayfish (Cherax cainii,Austin and Ryan 2002) aquaculture

Two experiments were conducted to evaluate the efficacy of an oil-based substrate, The Water Cleanser? (TWC). The first experiment studied the effects two substrates of different oil composition (TWC, TWC?+), and a commercial bacterial additive (Bio-Aid) on concentrations of nitrogen and phosphorous in indoor aquaria. The second experiment studied the effects of TWC, a bacterial additive, and a combination (TWC?+?B) on concentrations of nitrogen and phosphorous, phytoplankton abundance and diversity in outdoor freshwater crayfish (Cherax cainii, Austin and Ryan, Invertebr Syst 16:357–367, 2002) tanks. In the first experiment, the concentration of TAN decreased more rapidly with Bio-Aid, whilst the maximum concentrations of NO₂-N and NO₃-N were reduced with the substrates. The concentration of orthophosphate was reduced in aquaria with TWC?+?. In the second experiment, concentrations of TAN, NO₂-N, NO₃-N and total phosphate were not significantly affected by TWC. After addition of TWC?+?B, there was a significant decrease in the concentrations of nitrate and total phosphate, and a higher abundance of phytoplankton was maintained than with other treatments. Additionally, a population Bacillus sp. was found on the substrate surface. TWC had no adverse effects on phytoplankton abundance or diversity, or C. cainii physiology, weight gain or survival. TWC and TWC?+?were effective bioremediators of eutrophic water, whilst a combination of TWC?+?B was effective in short term bioremediation and in promoting phytoplankton abundance in C. cainii tank culture.