Genome-wide association studies and genomic selection assays made in a large sample of cacao (Theobroma cacao L.) germplasm reveal significant marker-trait associations and good predictive value for improving yield potential.

A genome-wide association study (GWAS) was undertaken to unravel marker-trait associations (MTAs) between SNP markers and phenotypic traits. It involved a subset of 421 cacao accessions from the large and diverse collection conserved ex situ at the International Cocoa Genebank Trinidad. A Mixed Linear Model (MLM) in TASSEL was used for the GWAS and followed by confirmatory analyses using GAPIT FarmCPU. An average linkage disequilibrium (r2) of 0.10 at 5.2 Mb was found across several chromosomes. Seventeen significant (P ≤ 8.17 × 10-5 (-log10 (p) = 4.088)) MTAs of interest, including six that pertained to yield-related traits, were identified using TASSEL MLM. The latter accounted for 5 to 17% of the phenotypic variation expressed. The highly significant association (P ≤ 8.17 × 10-5) between seed length to width ratio and TcSNP 733 on chromosome 5 was verified with FarmCPU (P ≤ 1.12 × 10-8). Fourteen MTAs were common to both the TASSEL and FarmCPU models at P ≤ 0.003. The most significant yield-related MTAs involved seed number and seed length on chromosome 7 (P ≤ 1.15 × 10-14 and P ≤ 6.75 × 10-05, respectively) and seed number on chromosome 1 (P ≤ 2.38 × 10-05), based on the TASSEL MLM. It was noteworthy that seed length, seed length to width ratio and seed number were associated with markers at different loci, indicating their polygenic nature. Approximately 40 candidate genes that encode embryo and seed development, protein synthesis, carbohydrate transport and lipid biosynthesis and transport were identified in the flanking regions of the significantly associated SNPs and in linkage disequilibrium with them. A significant association of fruit surface anthocyanin intensity co-localised with MYB-related protein 308 on chromosome 4. Testing of a genomic selection approach revealed good predictive value (genomic estimated breeding values (GEBV)) for economic traits such as seed number (GEBV = 0.611), seed length (0.6199), seed width (0.5435), seed length to width ratio (0.5503), seed/cotyledon mass (0.6014) and ovule number (0.6325). The findings of this study could facilitate genomic selection and marker-assisted breeding of cacao thereby expediting improvement in the yield potential of cacao planting material.

Switching the solubility of electroactive ionic liquids for designing high energy supercapacitor and low potential biosensor.

Ionic liquids are regarded as one of the most prodigious materials for sustainable technological developments with superior performance and versatility. Hence, in this study, we have reported the design and synthesis of electroactive disubstituted ferrocenyl ionic liquids (Fc-ILs) with two different counter anions and demonstrated the significance of their anion tuneable physicochemical characteristics towards multifunctional electrochemical applications. The Fc-IL synthesized with chloride counter anion (Fc-Cl-IL) displays water-solubility and can be used as a redox additive in the fabrication of supercapacitor. Supercapacitor device with Fc-Cl-IL based redox electrolyte exhibits outstanding energy and power densities of 91 Wh kg-1 and 20.3 kW kg-1, respectively. Meanwhile, ferrocenyl IL synthesized with perchlorate anion (Fc-ClO4-IL) exhibits water-insolubility and can serve as a redox mediator towards construction of a glucose biosensor. The biosensor comprising Fc-ClO4-IL is able to detect glucose at an exceptionally lower potential of 0.2 V, with remarkable sensitivity and selectivity. This study implies that the introduction of electroactive ILs could afford supercapacitor devices with high energy and power densities and biosensors with less detection potential.

Water insoluble, self-binding viologen functionalized ionic liquid for simultaneous electrochemical detection of nitrophenol isomers.

Herein, a water insoluble viologen functionalized ionic liquid (Vio-IL) was designed for the simultaneous electrochemical detection of 2-nitrophenol and 4-nitrophenol. Carboxyl functionalized benzimidazolium based ILs were hierarchically synthesized and the water insoluble carboxylic IL was covalently attached with aminopropylmethyl viologen through DCC coupling to obtain water insoluble Vio-IL gel. Vio-IL was immobilized by simple dropcasting over a multiwalled carbon nanotube deposited screen printed carbon electrode to obtain Vio-IL/MWCNT/SPE. The Vio-IL/MWCNT/SPE exhibited two sets of well-resolved redox peaks corresponding to V2+ to V+• and V+• to V0 redox couple. Furthermore, Vio-IL/MWCNT/SPE has shown excellent electrocatalytic activity towards the reduction of 2- and 4-nitrophenol individually, and also for the simultaneous determination of 2- and 4-nitrophenol. The linear range, sensitivity and detection limit for simultaneous detection of 2-nitrophenol at Vio-IL/MWCNT/SPE were found to be 4-494 µM, 0.884 µA µM-1 cm-2, 1.5 µM and that for 4-nitrophenol were 2-259 µM, 2.615 µA µM-1 cm-2, 0.70 µM. The Vio-IL/MWCNT/SPE established excellent sensitivity and selectivity towards the simultaneous determination of 2- and 4-nitrophenol together with impressive stability and reproducibility. The exemplary analytical parameters achieved are due to the rational immobilization of the mediator covalently in the highly conducting IL/MWCNT backbone which maintained the mediator characteristics effectively.

Rationally designed naphthyl substituted amine functionalized ionic liquid platform for covalent immobilization and direct electrochemistry of hemoglobin.

Herein, we have designed and demonstrated a facile and effective platform for the covalent anchoring of a tetrameric hemoprotein, hemoglobin (Hb). The platform comprises of naphthyl substituted amine functionalized gel type hydrophobic ionic liquid (NpNH2-IL) through which the heme protein was covalently attached over a glassy carbon electrode (Hb-NpNH2-IL/GCE). UV-vis and FT-IR spectral results confirmed that the Hb on NpNH2-IL retains its native structure, even after being covalently immobilized on NpNH2-IL platform. The direct electron transfer of redox protein could be realized at Hb-NpNH2-IL/GCE modified electrode and a well resolved redox peak with a formal potential of -0.30 V and peak separation of 65 mV was observed. This is due to the covalent attachment of highly conducting NpNH2-IL to the Hb, which facilitates rapid shuttling of electrons between the redox site of protein and the electrode. Further, the fabricated biosensor favoured the electrochemical reduction of bromate in neutral pH with linearity ranging from 12 to 228 µM and 0.228 to 4.42 mM with a detection limit and sensitivities of 3 µM, 430.7 µA mM-1 cm-2 and 148.4 µA mM-1 cm-2 respectively. Notably, the fabricated biosensor showed good operational stability under static and dynamic conditions with high selectivity and reproducibility.

Identification of Resistant Cultivars for Sheath Blight and use of AMMI Models to Understand Genotype and Environment Interactions.

One hundred and one rice genotypes were evaluated for response to sheath blight disease under greenhouse and lowland irrigated field conditions in Guyana. The level of resistance varied from highly resistant to resistant in 14 genotypes over five experimental trials. These genotypes were also observed with low area under the disease progress curve values and slow blighting reactions against artificial inoculation of the pathogen. Genotypes GR1568-31-9-1-1-2-1 and cultivar Rustic had susceptible reactions in all experiments. Additive main effect and multiplicative interaction analysis was used to study the genotype and environment interactions. The analysis revealed that 52.98% of the total sum of square was attributed to genotype effect, 7.50% was attributable to environment effect, and 39.52% was attributable to genotype by environment interaction (G × E) effects. The G × E was almost as large as the genotype effect, thus indicating significant differences of genotypes across the testing environments. This revealed that resistance was slightly influenced by the G × E. The genotypes that showed stable resistance in all environments in this study could be used for breeding the sheath blight resistance in rice.

Aldehyde functionalized ionic liquid on electrochemically reduced graphene oxide as a versatile platform for covalent immobilization of biomolecules and biosensing.

An aldehyde functionalized ionic liquid, (3-(3-formyl-4-hydroxybenzyl)-3-methylimidazolium hexafluorophosphate) (CHO-IL) has been employed herein as a multiple host platform for the covalent immobilization of mediator as well as enzyme. The CHO-IL was immobilized on electrochemically reduced graphene oxide (ERGO) through the π-π stacking of hydroxybenzyl and imidazolium groups with ERGO and subjected to further covalent attachment of Azure A (Azu-A) mediator or glucose oxidase (GOx) enzyme. Electroactive, water soluble organic dye Azu-A was effectively immobilized to the host IL through simple Schiff base reaction. Azu-A was rendered leak-free in the electrode setup and also responded well for the amperometric determination of H2O2 over a linear range of 0.03-1mM with a detection limit and sensitivity of 11.5µM and 133.2µAmM-1cm-2, respectively. Further, attempts were made to explore the CHO-IL platform for the covalent immobilization of GOx enzyme which served well in retaining the enzyme nativity, reactivity and stability. Under optimized conditions, mediatorless GOx biosensor developed based on direct electrochemistry has exhibited an impressive analytical signal towards glucose detection in the linear range of 0.05-2.4mM with a detection limit and sensitivity of 17µM and 17.7µAmM-1cm-2, respectively. The reliability of the proposed Azu-A based chemical sensor and GOx based biosensor towards the determination of H2O2 and glucose in the real samples have been demonstrated. The remarkable analytical parameters and long term stability of both the sensors could be envisioned as a result of facile immobilization platform and immobilization strategy.

Electrocatalytic Conversion of Carbon Dioxide and Nitrate Ions to Urea by a Titania-Nafion Composite Electrode.

CO2 and nitrate ions were successfully converted to urea by a TiO2 -Nafion nanocomposite electrode under ambient conditions. The composite electrode was constructed by dropcasting the mixture of P25 titania and Nafion solution on an indium-doped tin oxide (ITO) electrode. When the electrode was electrolyzed in CO2 -saturated 0.1 m KNO3 (pH 4.5) solution at -0.98 V versus Ag/AgCl, urea was formed with a Faradaic efficiency of 40 %. The other reduced products obtained were NH3 , CO, and H2 .

A bioinspired ionic liquid tagged cobalt-salophen complex for nonenzymatic detection of glucose.

The development of efficient and cost effective nonenzymatic biosensors with remarkable sensitivity, selectivity and stability for the detection of biomolecules, especially glucose is one of the major challenges in materials- and electrochemistry. Herein, we report the design and preparation of nonenzymatic biosensor based on an ionic liquid tagged cobalt-salophen metal complex (Co-salophen-IL) immobilized on electrochemically reduced graphene oxide (ERGO) for the detection of glucose via an electrochemical oxidation. The bioinspired Co-salophen-IL complex has been synthesized and immobilized on ERGO, which was previously deposited on a screen printed carbon electrode (SPE) to form the Co-salophen-IL/ERGO/SPE nonenzymatic biosensor. The electrochemical behaviour of this modified electrode was studied using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Notably, the Co-salophen-IL/ERGO/SPE biosensor exhibited excellent electrocatalytic activity towards glucose oxidation in 0.1M NaOH, based on which an amperometric sensor has been developed. The modified electrode has shown prominent performance towards glucose detection over a wide linear range from 0.2µM to 1.8mM with a detection limit and sensitivity of 0.79µM and 62µAmM-1 respectively. The detection was carried out at 0.40V and such a less working potential excludes the interference from the coexisting oxidizable analytes. The role of Co-salophen, IL and ERGO in the electrocatalytic activity has been systematically investigated. Furthermore, the biosensor demonstrated high stability with good reproducibility.

Combination of endophytic Bacillus and Beauveria for the management of Fusarium wilt and fruit borer in tomato.

BACKGROUND: Most of the approaches for biocontrol of pests and diseases have used a single biocontrol agent as antagonist to a single pest or pathogen. This accounts for the inconsistency in the performance of biocontrol agents. The development of a bioformulation possessing a mixture of bioagents could be a viable option for the management of major pests and diseases in crop plants. RESULTS: A bioformulation containing a mixture of Beauveria bassiana (B2) and Bacillus subtilis (EPC8) was tested against Fusarium wilt and fruit borer in tomato under glasshouse and field conditions. The bioformulation with B2 and EPC8 isolates effectively reduced the incidence of Fusarium wilt (Fusarium oxysporum f. sp. lycopersici) and fruit borer (Helicoverpa armigera) under glasshouse and field conditions compared with the individual application of B2 and EPC8 isolates and control treatments. In vitro studies showed a higher larval mortality of H. armigera when fed with B2 + EPC8-treated leaves. Further, plants treated with the B2 + EPC8 combination showed a greater accumulation of defence enzymes such as lipoxygenase, peroxidase and polyphenol oxidase against wilt pathogen and fruit borer pest than the other treatments. Moreover, a significant increase in growth parameters and yield was observed in tomato plants treated with B2 + EPC8 compared with the individual bioformulations and untreated control. CONCLUSION: The combined application of Beauveria and Bacillus isolates B2 and EPC8 effectively reduced wilt disease and fruit borer attack in tomato plants. Results show the possibility of synchronous management of tomato fruit borer pest and wilt disease in a sustainable manner.

Reduction of CO2 to CO at low overpotential in neutral aqueous solution by a Ni(cyclam) complex attached to poly(allylamine).

Less is more: Nickel cyclam complexes are known as efficient catalysts for the electrochemical reduction of CO(2) to CO, despite having overvoltages of more than 0.6 V. Incorporating [Ni(cyclam)](2+) into poly(allylamine) through axial coordination of pyridine enables the electrochemical reduction of CO(2) to CO near the thermodynamic potential of -0.78 V (vs. Ag/AgCl) at pH 8 with a current efficiency of 90 %.

PGPR and entomopathogenic fungus bioformulation for the synchronous management of leaffolder pest and sheath blight disease of rice.

BACKGROUND: The biological control of plant pests and diseases using a single organism has been reported to give inconsistent and poor performance. To improve the efficacy, bioformulations were developed possessing mixtures of bioagents. RESULTS: Bioformulations combining Pseudomonas fluorescens Migula strains Pf1 and AH1 and Beauveria bassiana (Balsamo) Vuill. isolate B2 were developed and tested for their efficacy against leaffolder pest and sheath blight disease on rice under glasshouse and field conditions. The combination of Pf1, AH1 and B2 effectively reduced the incidence of leaffolder insect and sheath blight disease on rice compared with other treatments. An in vitro assay of leaffolder preference to rice leaf tissues treated with Pf1 + AH1 + B2 biformulation showed variation from normal growth and development of leaffolder larvae. Plants treated with the Pf1 + AH1 + B2 combination showed a greater accumulation of enzymes, lipoxygenase and chitinase activity against leaffolder insect compared with other treatments. Similarly, the plants showed a higher accumulation of defence enzymes, peroxidase and polyphenol oxidase activity against sheath blight pathogen in Pf1 + AH1 + B2 treatment compared with the untreated control. The bioformulation mixture attracted the natural enemy population of leaffolder under field conditions. In addition, a significant increase in rice grain yield was observed in Pf1 + AH1 + B2 treatment compared with the untreated control. CONCLUSION: The combination of P. fluorescens strains and B. bassiana isolate effectively reduced the incidence of leaffolder insect and sheath blight disease on rice plants and showed the possibility of controlling both pest and disease using a single bioformulation.

Pseudomonas-induced defence molecules in rice plants against leaffolder (Cnaphalocrocis medinalis) pest.

Fluorescent pseudomonad strains Pf1, TDK1 and PY15 individually and in combination were evaluated against leaffolder, Cnaphalocrocis medinalis Guen., in rice under in vitro, glasshouse and field conditions. Among the various treatments used, a combination of Pf1, TDK1 and PY15 strains effectively reduced the incidence of leaffolder pest in rice plants to an extent comparable with chlorpyrifos-methyl. In addition, morphogenesis of the insect pest in all stages, larval, pupal and adult, was greatly affected by a combination of Pseudomonas Pf1, TDK1 and PY15 strains. Further, the induction of defence-related molecules was demonstrated. An increased accumulation of defence molecules such as chitinase and proteinase inhibitors was observed with a combined Pf1, TDK1 and PY15 treatment compared with all other treatments. Western blot analysis of chitinase revealed the extra induction of 18, 28 and 35 kDa isoforms in rice plants treated with a mixture of Pf1, TDK1 and PY15 strains against leaffolder pest. The study revealed that a combination of fluorescent pseudomonad strains affects the development of leaffolder pest by inducing defence molecules in rice plants which in turn enhance resistance to leaffolder attack.