Functional Coupling of a Nematode Chemoreceptor to the Yeast Pheromone Response Pathway

Sequencing of the Caenorhabditis elegans genome revealed sequences encoding more than 1,000 G-protein coupled receptors, hundreds of which may respond to volatile organic ligands. To understand how the worm''s simple olfactory system can sense its chemical environment there is a need to characterise a representative selection of these receptors but only very few receptors have been linked to a specific volatile ligand. We therefore set out to design a yeast expression system for assigning ligands to nematode chemoreceptors. We showed that while a model receptor ODR-10 binds to C. elegans Gα subunits ODR-3 and GPA-3 it cannot bind to yeast Gα. However, chimaeras between the nematode and yeast Gα subunits bound to both ODR-10 and the yeast Gβγ subunits. FIG2 was shown to be a superior MAP-dependent promoter for reporter expression. We replaced the endogenous Gα subunit (GPA1) of the Saccharomyces cerevisiae (ste2Δ sst2Δ far1Δ) triple mutant (“Cyb”) with a Gpa1/ODR-3 chimaera and introduced ODR-10 as a model nematode GPCR. This strain showed concentration-dependent activation of the yeast MAP kinase pathway in the presence of diacetyl, the first time that the native form of a nematode chemoreceptor has been functionally expressed in yeast. This is an important step towards en masse de-orphaning of C. elegans chemoreceptors.     

Foliar application of melatonin delay leaf senescence in maize by improving the antioxidant defense system and enhancing photosynthetic capacity under semi-arid regions

Melatonin is an important plant growth regulator which plays a key role in plant growth and development. The objective of the current research was to evaluate the effect of foliar application of melatonin (MF) on photosynthetic efficiency, antioxidant defense mechanism, and its relation with leaf senescence in maize crop grown in a semi-arid region. A field experiment was conducted during 2017 and 2018 growth season, where melatonin was applied to the foliage at concentrations of 0 (MF₀), 25 (MF₁), 50 (MF₂), and 75 (MF₃) μM at the ninth leaf stage. Foliar application of melatonin significantly improved chlorophyll content, net photosynthetic rate, soluble sugar content, and soluble protein content during the process of leaf senescence. The application of melatonin also enhanced antioxidant enzyme activities including superoxide dismutase, catalase, and peroxidase, while reduced malondialdehyde and reactive oxygen species accumulation. Melatonin foliar application also increased total leaf area per plant, grains per ear, thousand grain weight and grain yield of maize crop in a semi-arid region. The application of melatonin significantly improved photosynthetic activity, antioxidant defense mechanism, and yield of maize crop in a semi-arid region, where the most effective treatment was MF2.

     

Seed priming improves the emergence potential,growth and antioxidant system of Moringa oleifera under saline conditions

Moringa oleifera is a multipurpose plant which is now being promoted as a fodder crop. The present study was conducted to induce the tolerance in moringa plants to emerge and grow under saline conditions. For this, moringa seeds were primed with aerated water (hydropriming) and moringa leaf extract (MLE) for 12 and 24 h and studied for its emergence, potential growth behaviour, mineral composition, chlorophyll contents and antioxidant activities in comparison with unprimed seeds to investigate the physiological changes in moringa plants under saline conditions. The seeds were sown in plastic pots filled with acid washed sand at four salinity levels (3, 6, 10, 14 dS m^?1) in a completely randomized design with three replications. It was found that salinity >6 dS m^?1 reduced the emergence, growth and vigour of moringa plants but hydropriming (12 h) enhanced moringa emergence at 10 dS m^?1 followed by MLE priming (12 h). Maximum aboveground biomass and photosynthetic pigments were recorded when the seeds were hydroprimed (12 h) but maximum root length and number of roots were found in MLE primed (12 h) moringa plants. Significant decrease in K⁺:Na⁺ ratio with increasing salinity levels resulted in low K⁺ and Mg²⁺ uptake and Na⁺ toxicity in moringa leaves which resulted in reduced chlorophyll contents at 14 dS m^?1 but a significant increase in chlorophyll a and b contents and total phenolics were found in hydroprimed seeds (12 h) while the antioxidant activities of superoxide dismutase, peroxidase and catalas were improved by MLE priming (12 h). This study concludes that moringa emergence and growth performance can be improved by hydropriming under saline conditions.     

TSGIT: An N‐ and C‐terminal tandem tag system for purification of native and intein‐mediated ligation‐ready proteins

A large variety of fusion tags have been developed to improve protein expression, solubilization, and purification. Nevertheless, these tags have been combined in a rather limited number of composite tags and usually these composite tags have been dictated by traditional commercially‐available expression vectors. Moreover, most commercially‐available expression vectors include either N‐ or C‐terminal fusion tags but not both. Here, we introduce TSGIT, a fusion‐tag system composed of both N‐ and a C‐terminal composite fusion tags. The system includes two affinity tags, two solubilization tags and two cleavable tags distributed at both termini of the protein of interest. Therefore, the N‐ and the C‐terminal composite fusion tags in TSGIT are fully orthogonal in terms of both affinity selection and cleavage. For using TSGIT, we streamlined the cloning, expression, and purification procedures. Each component tag is selected to maximize its benefits toward the final construct. By expressing and partially purifying the protein of interest between the components of the TSGIT fusion, the full‐length protein is selected over truncated forms, which has been a long‐standing problem in protein purification. Moreover, due to the nature of the cleavable tags in TSGIT, the protein of interest is obtained in its native form without any additional undesired N‐ or C‐terminal amino acids. Finally, the resulting purified protein is ready for efficient ligation with other proteins or peptides for downstream applications. We demonstrate the use of this system by purifying a large amount of native fluorescent mRuby3 protein and bacteriophage T7 gp2.5 ssDNA‐binding protein.     

Metallochaperone-like genes in Arabidopsis thaliana

A complete inventory of metallochaperone-like proteins containing a predicted HMA domain in Arabidopsis revealed a large family of 67 proteins. 45 proteins, the HIPPs, have a predicted isoprenylation site while 22 proteins, the HPPs, do not. Sequence comparisons divided the proteins into seven major clusters (I-VII). Cluster IV is notable for the presence of a conserved Asp residue before the CysXXCys, metal binding motif, analogous to the Zn binding motif in E. coli ZntA. HIPP20, HIPP21, HIPP22, HIPP26 and HIPP27 in Cluster IV were studied in more detail. All but HIPP21 could rescue the Cd-sensitive, ycf1 yeast mutant but failed to rescue the growth of zrt1zrt2, zrc1cot1 and atx1 mutants. In Arabidopsis, single and double mutants did not show a phenotype but the hipp20/21/22 triple mutant was more sensitive to Cd and accumulated less Cd than the wild-type suggesting the HIPPs can have a role in Cd-detoxification, possibly by binding Cd. Promoter-GUS reporter expression studies indicated variable expression of these HIPPs. For example, in roots, HIPP22 and HIPP26 are only expressed in lateral root tips while HIPP20 and HIPP25 show strong expression in the root vasculature.     

Distinct Phenotypes Caused by Mutation of MSH2 in Trypanosome Insect and Mammalian Life Cycle Forms Are Associated with Parasite Adaptation to Oxidative Stress

BackgroundDNA repair mechanisms are crucial for maintenance of the genome in all organisms, including parasites where successful infection is dependent both on genomic stability and sequence variation. MSH2 is an early acting, central component of the Mismatch Repair (MMR) pathway, which is responsible for the recognition and correction of base mismatches that occur during DNA replication and recombination. In addition, recent evidence suggests that MSH2 might also play an important, but poorly understood, role in responding to oxidative damage in both African and American trypanosomes.Conclusions/SignificanceTaken together, these results indicate MSH2 displays conserved, dual roles in MMR and in the response to oxidative stress. Loss of the latter function results in life cycle dependent differences in phenotypic outcomes in T. brucei MSH2 mutants, most likely because of the greater burden of oxidative stress in the insect stage of the parasite.     

Investigation of Plasmonic Bandgap for 1D Exposed and Buried Metallic Gratings

Plasmonics - A simulation study for the opening of plasmonic bandgap (PBG) with control over it by varying the slit width (SW) for exposed and buried 1D metallic gratings has been reported by using...     

Cross-linked stem bromelain: A more stabilized active preparation

Cross-linking of the protease stem bromelain (bromelain) with 0.25 and 1.25% glutaraldehyde (GTA) results in the formation of a large molecular mass, multimeric and soluble aggregate having comparable activity to the unmodified bromelain. Both 0.25 and 1.25% GTA cross-linked (CL) bromelain preparations were more stable against urea, guanidine hydrochloride (GdnHCl) and temperature-induced inactivation, and exhibited slightly better storage stability compared to the unmodified protease. Such a high molecular weight, soluble, active and stable preparation may be useful in industry, i.e. in the textile industry for improving the properties of a fabric without loss of fabric strength and shape.     

Oligomerisation of C. elegans Olfactory Receptors,ODR-10 and STR-112, in Yeast

It is widely accepted that vertebrate G-Protein Coupled Receptors (GPCRs) associate with each other as homo- or hetero-dimers or higher-order oligomers. The C. elegans genome encodes hundreds of olfactory GPCRs, which may be expressed in fewer than a dozen chemosensory neurons, suggesting an opportunity for oligomerisation. Here we show, using three independent lines of evidence: co-immunoprecipitation, bioluminescence resonance energy transfer and a yeast two-hybrid assay that nematode olfactory receptors (ORs) oligomerise when heterologously expressed in yeast. Specifically, the nematode receptor ODR-10 is able to homo-oligomerise and can also form heteromers with the related nematode receptor STR-112. ODR-10 also oligomerised with the rat I7 OR but did not oligomerise with the human somatostatin receptor 5, a neuropeptide receptor. In this study, the question of functional relevance was not addressed and remains to be investigated.     

Optimum leaf excision increases the biomass accumulation and seed yield of maize plants under different planting patterns

Without developing new agronomic practices, present rates of improvement in seed yields of cereal crops globally are insufficient to fulfil the estimated increasing food demand for 2050 and beyond. Intercropping is one of the agricultural practices that can lead to greater crop yields. However, there exists leaf redundancy for maize in intercropping systems, and the top canopy leaves shade more competent leaves at middle strata of maize plants. Therefore, this work aimed to elucidate the effect of leaf excision treatments in maize to understand the optimum leaf area of maize plants under a maize–soybean relay‐intercropping system (MS_R) and a sole cropping system (SM). The effects of four‐leaf excision treatments (T₁, 0; T₂, 2; T₃, 4; T₄, 6 leaves excised from the top of maize plants until 7 days after silking) on light interception, leaf area index (LAI), photosynthetic characteristics, total biomass accumulation at blistering stage (BS), dough stage (DS) and physiological maturity (PM), and seed yield of maize were investigated through field experiments for 2 years under MS_R and SM. Results showed that, under MS_R and SM, as compared to control (T₁), optimum excision of leaves (T₂) from the top of maize plants significantly improved the light interception (by 25, 18 and 16% at BS, DS and PM, respectively) to lower strata leaves and accelerated the biomass partitioning to maize seeds (by 13 and 12% at DS and PM, respectively). Importantly, plants under T₂ exhibited higher green leaf area than control, that is, excision the top two leaves led to an increase in LAI at PM by 10%, suggesting that leaf senescence under T₂ was delayed which enhanced the photosynthetic rate at PM by 7% in 2017 and 6% in 2018. Relative to T₁, maize under T₂ produced 19 and 13% higher maize yield under MS_R and SM, respectively, and relay‐cropped maize had 90% of SM seed yield. These results suggest that by manipulating the canopy structure of maize plants we can enhance the biomass accumulation and seed yield of maize crops under MS_R and SM.