Genetic variation among selected pure lines from Turkish barley landrace ‘Tokak’ in yield-related and malting quality traits

Aim of study: Improvement of barley cultivars for malting traits suffers from narrow genetic pool in barley for these traits. Landraces are resources that could be used for this purpose. The present study was conducted to determine the variation for malting quality traits within a Turkish barley landrace. Area of study: The study was undertaken in Tokat, a province in Black Sea Region of Turkey. Material and methods: Twenty-five diverse lines, out of 42 unique genotypes previously identified in ‘Tokak’ landrace (PI 470281) based on DNA markers, were evaluated for malting quality traits along with the malting barley cv. ‘Tokak 157/37’ in four field trials. Thousand-seed weight, test weight, grain yield, lodging, malt extract percentage, diastatic power, alpha amylase and malt beta glucanase activities, malt protein and starch contents were determined. Main results: Principal component analysis of malting quality traits revealed that thousand-seed weight, alpha amylase activity, beta glucanase activity and diastatic power were the most discriminatory traits for the lines. As the average of four trials, 15 of the 25 lines evaluated had higher grain yields and 10 of 25 lines had higher malt extract percentages than the standard cultivar ‘Tokak 157/37’. Malt extract was highest in Line 59 in all environments, and this line also had the highest values for beta glucanase activity and starch content. Line 215 had highest values for alpha amylase activity. Lines 59 and 215 clearly had superior malting quality. Research highlights: These lines could harbor novel alleles for these traits to be used in malting barley improvement.

Evaluation of Cellulolytic Endo-1,4-β-D-Glucanase Activity in the Digestive Fluid of Adult Phytophagous Beetle Hoplasoma unicolor

Insects of the taxonomic order Coleoptera are recognised for considerable cellulolytic activity in their digestive fluid. The cellulolytic activity of the gut fluid in Hoplasoma unicolor, a member of Coleoptera, however, remains unexplored. In this study, we, for the first time, report the qualitative and quantitative analysis of cellulolytic activity in the digestive fluid of this insect. The cellulolytic endo-1,4-β-D-glucanase activity was confirmed in the supernatant of the insect’s digestive fluid by agar plate assay using carboxymethyl cellulose as the substrate. To determine the optimum pH, enzyme activity was further assessed in an acidic pH range of 5 to 6, and the highest activity was observed at pH 5.3. For quantitative analysis, endoglucanase activity was measured using 3,5-dinitrosalicylic acid method which revealed that the specific activity of the gut sample was 0.69 (±0.01) units per mg of protein. For further characterisation of the cellulases in the sample, SDS-PAGE and zymogram analysis were carried out. Two active cellulolytic bands were detected on the zymogram suggesting the presence of two distinct endoglucanases which completely disappeared upon heating the sample at 55°C. Our study, therefore, highlights prospect of the gut fluid of H. unicolor as an important source of cellulase enzymes that merits further investigations into their extensive characterisation for potential industrial applications.

Development of Active Packaging to Extend the Shelf Life of Agaricus bisporus by Using Plasma Technology

In this study, a preservation package that can extend the shelf life of Agaricus bisporus was developed using plasma modification combined with low-density polyethylene (LDPE), collagen (COL), and carboxymethyl cellulose (CMC). Out results showed that the selectivity of LDPE to gas can be controlled by plasma modification combined with coating of different concentrations of CMC and COL. Packaging test results applied to A. bisporus showed that 3% and 5% of CMC and COL did not significantly inhibit polyphenol oxidase and β-1,3-glucanase, indicating no significant effect on structural integrity and oxidative browning. The use of 0.5% and 1.0% CMC and COL can effectively inhibit the polyphenol oxidase and β-1,3-glucanase activity of A. bisporus, leading to improved effects in browning inhibition and structural integrity maintenance. P-1.0COL can effectively maintain gas composition in the package (carbon dioxide: 10–15% and oxygen: 8–15%) and catalase activity during storage, thereby reducing the oxidative damage caused by respiration of A. bisporus. The current study confirmed that the use of plasma modification technology combined with 1.0% COL can be used in preservation packaging by regulating the respiration of A. bisporus, thus extending its shelf life from 7 to 21 days.

Biocontrol Potential of Neem Leaf-Based Vermicompost as Indicated by Chitinase, Protease and Β-1,3-Glucanase Activity

The rising concern regarding the negative impact of synthetic pesticides has led to the search for alternative means of pest control. Vermicomposting the mixture of oil palm empty fruit bunch and neem (Azadirachta indica) leaves, with the latter known to have pesticidal value, is therefore of great interest and significance to be studied. The present study was conducted to evaluate the chitinase, protease and β-1,3-glucanase activity of neem leaf-based vermicompost as an indication of its biocontrol properties. The total microbial population of different composition of the vermicompost was also investigated. The results showed that at 10% neem composition, an increment in microbial population, chitinase and protease activities was observed in the end product. A higher concentration of neem exerted a suppressive effect on the microbial population as well as enzymatic activity. This study suggested that the addition of an appropriate composition of neem leaves as one of the raw materials for vermicomposting would potentially enhance the performance of vermicompost as biofertilizer as well as biopesticide.

Arcopilus aureus MaC7A as a New Source of Resveratrol: Assessment of Amino Acid Precursors, Volatiles, and Fungal Enzymes for Boosting Resveratrol Production in Batch Cultures

The chemical factors that regulate the synthesis of resveratrol (RV) in filamentous fungi are still unknown. This work reports on the RV production by Arcopilus aureus MaC7A under controlled conditions and the effect of amino acid precursors (PHE and TYR), monoterpenes (limonone, camphor, citral, thymol, menthol), and mixtures of hydrolytic enzymes (Glucanex) as elicitors for boosting fungal RV. Batch cultures with variable concentrations of PHE and TYR (50–500 mg L−1) stimulated RV production from 127.9 ± 4.6 to 221.8 ± 5.2 mg L−1 in basic cultures developed in PDB (pH 7) added with 10 g L−1 peptone at 30 °C. Maximum levels of RV and biomass were maintained during days 6–8 under these conditions, whereas a dramatic RV decrease was observed from days 10–12 without any loss of biomass. Among the tested volatiles, citral (50 mg L−1) enhanced RV production until 187.8 ± 2.2 mg L−1 in basic cultures, but better results were obtained with Glucanex (100 mg L−1; 198.3 ± 7.6 mg L−1 RV). Optimized batch cultures containing TYR (200 mg L−1), citral (50 mg L−1), thymol (50 mg L−1), and Glucanex (100 mg L−1) produced up to 237.6 ± 4.7 mg L−1 of RV. Our results suggest that low concentrations of volatiles and mixtures of isoenzymes with β-1, 3 glucanase activity increase the biosynthesis of fungal RV produced by A. aureus MaC7A in batch cultures.

Fed-Batch Cultivation and Adding Supplements to Increase Yield of β-1,3-1,4-Glucanase by Genetically Engineered Escherichia coli

The aim of this study was to analyze the major influence factors of culture medium on the expression level of β-1,3-1,4-glucanase, and to further develop an optimized process for the extracellular production of β-glucanase at a bioreactor scale (7 L) with a genetically engineered Escherichia coli (E. coli) JM109-pLF3. In this study, batch cultivation and fed-batch cultivation including the constant rate feeding strategy and the DO-stat (DO: Dissolved Oxygen) feeding strategy were conducted. At a 7 L bioreactor scale for batch cultivation, biomass reached 3.14 g/L and the maximum β-glucanase activity was 506.94 U/mL. Compared with batch cultivation, the addition of glycerol, complex nitrogen and complete medium during fed-batch cultivation increased the production of biomass and β-1,3-1,4-glucanase. The maximum biomass and β-glucanase activity, which were 7.67 g/L and 1680 U/mL, respectively, that is, 2.45 and 3.31 times higher than those obtained with batch cultivation, were obtained by feeding a complex nitrogen source at a constant rate of 1.11 mL/min. Therefore, these nutritional supplements and strategies can be used as a reference to enhance the production of other bioproducts from E. coli.

Application of a robust microplate assay to determine induced β-1,3-glucanase and chitinase activity in the cotton plant

Resistance is induced in cotton plants as the result of either viral infection or exogenous application of elicitors. Induced resistance can be evaluated by determining the production of β-1,3-glucanase and chitinase in plants as a biochemical parameter. The assays being used for the determination of chitinase and β-1,3-glucanase activity are laborious and not cost-effective, as the reducing sugars produced by the substrates colloidal chitin and laminarin are very expensive. The concentration of both substrates was standardized and reduced to 0.25% from 4% in a modified microplate assay, which appeared to be more effective. The amount of β-1,3-glucanase and chitinase produced was significant and determined by the new modified assay. The sensitivity of the microplate assay was significantly raised approximately one- to twofold.

Epiphytic fungi induced pathogen resistance of invasive plant Ipomoea cairica against Colletotrichum gloeosporioides

Background Ipomoea cairica (L.) Sweet is a destructive invasive weed in South China but rarely infected with pathogens in nature. Its pathogen resistance mechanism is largely unknown at present. Some non-pathogenic isolates of Fusarium oxysporum and Fusarium fujikuroi are prevalent on many plant species and function as pathogen resistance inducers of host plants. The objective of the present research is to investigate whether the symbiosis between the both fungi and I. cairica is present, and thereby induces pathogen resistance of I. cairica. Methods Through field investigation, we explored the occurrence rates of F. oxysporum and F. fujikuroi on leaf surfaces of I. cairica plants in natural habitats and compared their abundance between healthy leaves and leaves infected with Colletotrichum gloeosporioides, a natural pathogen. With artificial inoculation, we assessed their pathogenicity to I. cairica and studied their contribution of pathogen resistance to I. cairica against C. gloeosporioides. Results We found that F. oxysporum and F. fujikuroi were widely epiphytic on healthy leaf surfaces of I. cairica in sunny non-saline, shady non-saline and sunny saline habitats. Their occurrence rates reached up to 100%. Moreover, we found that the abundance of F. oxysporum and F. fujikuroi on leaves infected with C. gloeosporioides were significantly lower than that of healthy leaves. With artificial inoculation, we empirically confirmed that F. oxysporum and F. fujikuroi were non-pathogenic to I. cairica. It was interesting that colonization by F. fujikuroi, F. oxysporum alone and a mixture of both fungi resulted in a reduction of C. gloeosporioides infection to I. cairica accompanied by lower lesion area to leaf surface area ratio, increased hydrogen peroxide (H2O2) concentration and salicylic acid (SA) level relative to the control. However, NPR1 expression, chitinase and β-1,3-glucanase activities as well as stem length and biomass of I. cairica plant only could be significantly improved by F. oxysporum and a mixture of both fungi but not by F. fujikuroi. In addition, as compared to colonization by F. oxysporum and a mixture of both fungi, F. fujikuroi induced significantly higher jasmonic acid (JA) level but significantly lower β-1,3-glucanase activity in leaves of I. cairica plants. Thus, our findings indicated the symbiosis of epiphytic fungiF. fujikuroi and F. oxysporum induced systemic resistance of I. cairica against C. gloeosporioides. F. oxysporum played a dominant role in inducing pathogen resistance of I. cairica. Its presence alleviated the antagonism of the JA signaling on SA-dependent β-1,3-glucanase activity and enabled I. cairica plants to maintain relatively higher level of resistance against C. gloeosporioides.

The antifungal peptide CGA-N12 inhibits cell wall synthesis of Candida tropicalis by interacting with KRE9

CGA-N12, an antifungal peptide derived from chromogranin A, has specific antagonistic activity against Candida spp., especially against Candida tropicalis, by inducing cell apoptosis. However, the effect of CGA-N12 on the Candida cell wall is unknown. The Candida protein KRE9, which possesses β-1,6-glucanase activity, was screened by affinity chromatography after binding to CGA-N12. In this study, the effect of CGA-N12 on KRE9 and the interaction between CGA-N12 and KRE9 was studied to clarify the effect of CGA-N12 on C. tropicalis cell wall synthesis. The effect of CGA-N12 on recombinant KRE9 β-1,6-glucanase activity was investigated by analyzing the consumption of glucose. The results showed that CGA-N12 inhibited the activity of KRE9. After C. tropicalis was treated with CGA-N12, the structure of the C. tropicalis cell wall was damaged. The interaction between CGA-N12 and KRE9 was analyzed by isothermal titration calorimetry (ITC). The results showed that their interaction process was involved an endothermic reaction, and the interaction force was mainly hydrophobic with a few electrostatic forces. The results of the fluorescence resonance energy transfer (FRET) assay showed that the distance between CGA-N12 and KRE9 was 7 ∼ 10 nm during their interaction. Therefore, we concluded that the target of CGA-N12 in the C. tropicalis cell membrane is KRE9, and that CGA-N12 weakly binds to KRE9 within a 7 ∼ 10 nm distance and inhibits KRE9 activity.

Epiphytic fungi induced pathogen resistance of invasive plant Ipomoea cairica against Colletotrichum gloeosporioides

Background. Ipomoea cairica (L.) Sweet is a destructive invasive weed in South China but rarely infected with pathogens in nature. Its pathogen resistance mechanism is largely unknown at present. Some non-pathogenic isolates of Fusarium oxysporum and Fusarium fujikuroi are prevalent on many plant species and function as pathogen resistance inducers of host plants. The objective of the present research is to investigate whether the symbiosis between the both fungi and I. cairica is present, and thereby induce pathogen resistance of I. cairica. Methods. Through field investigation, we explored the occurrence rates of F. oxysporum and F. fujikuroi on leaf surfaces of I. cairica plants in natural habitats and compared their abundance between healthy leaves and leaves infected with Colletotrichum gloeosporioides, a natural pathogen. With artificial inoculation, we assessed their pathogencity to I. cairica and study their contribution of pathogen resistance to I. cairica against C. gloeosporioides. Results. We found that F. oxysporum and F. fujikuroi were widely epiphytic on healthy leaf surfaces of I. cairica in sunny non-saline, shady non-saline and sunny saline habitats. Their occurrence rates reached up to 100%. Moreover, we found that the abundance of F. oxysporum and F. fujikuroi on leaves infected with C. gloeosporioides were significantly lower than that of healthy leaves. With artificial inoculation, we empirically confirmed that F. oxysporum and F. fujikuroi were non-pathogenic to I. cairica. It was interesting that colonization by F. fujikuroi, F. oxysporum alone and a mixture of both fungi resulted in a reduction of C. gloeosporioides infection to I. cairica accompanied by lower lesion area to leaf surface area ratio, increased H2O2 concentration and salicylic acid (SA) level relative to the control. However, NPR1 expression, chitinase and β -1,3-glucanase activities as well as stem length and biomass of I. cairica plant only could be significantly improved by F. oxysporum and a mixture of both fungi but not by F. fujikuroi. In addition, as compared to colonization by F. oxysporum and a mixture of both fungi, F. fujikuroi induced significantly higher jasmonic acid (JA) level but significantly lower β -1,3-glucanase activity in leaves of I. cairica plants. Thus, our findings indicated the symbiosis of epiphytic fungi F. fujikuroi and F. oxysporum facilitated the fitness of I. cairica via the induced systemic resistance of host plant against C. gloeosporioides. F. oxysporum played a dominant role in inducing pathogen resistance of I. cairica. Its presence alleviated the antagonism of the JA signaling on SA-dependent β -1,3-glucanase activity and enabled I. cairica plants to maintain relatively higher level of resistance against C. gloeosporioides.