Accumulation of Health-Promoting Compounds in Upland Black Rice by Interacting Mycorrhizal and Endophytic Fungi.

There is an increasing interest in finding eco-friendly and safe approaches to increase agricultural productivity and deliver healthy foods. Arbuscular mycorrhizal fungi (AMF) and endophytic fungi (EPF) are important components of sustainable agriculture in view of their ability to increase productivity and various plant secondary metabolites with health-promoting effects. In a pot experiment, our main research question was to evaluate the additive and synergistic effects of an AMF and four root-endophytic fungi on plant performance and on the accumulation of health-promoting secondary compounds. Plant growth varied between the treatments with both single inoculants and co-inoculation of an AMF and four EPF strains. We found that inoculation with a single EPF positively affected the growth and biomass production of most of the plant-endophyte consortia examined. The introduction of AMF into this experiment (dual inoculation) had a beneficial effect on plant growth and yield. AMF, Rhizophagus variabilis KS-02 co-inoculated with EPF, Trichoderma zelobreve PBMP16 increased the highest biomass, exceeding the growth rate of non-inoculated plants. Co-inoculated R. variabilis KS-02 and T. zelobreve PBMP16 had significantly greater beneficial effects on almost all aspects of plant growth, photosynthesis-related parameters, and yield. It also promoted root growth quality and plant nutrient uptake. The phenolic compounds, anthocyanin, and antioxidant capacity in rice seeds harvested from plants co-inoculated with AMF and EPF were dramatically increased compared with those from non-inoculated plants. In conclusion, our results indicated that EPF and AMF contributed to symbiosis in Maled Phai cultivar and were coordinately involved in promoting plant growth performance under a pot trial.

The Potential of Endophytic Fungi for Enhancing the Growth and Accumulation of Phenolic Compounds and Anthocyanin in Maled Phai Rice (Oryza sativa L.).

Rice (Oryza sativa L.) is one of the most popular cereal crops, being consumed by almost half of the world's population. Among several cultivars widely distributed in Thailand, Maled Phai is a Thai pigmented-upland rice with exceptionally high nutritional value and high demand in the local Thai market. This study aimed to investigate the feasibility of producing plant growth-promoting properties (PGP) and enhancing the accumulation of phytochemicals in Maled Phai rice seeds of endophytic fungi isolated from upland black rice. Among a total of 56 isolates, the 4 most effective PGP isolates were identified as Trichoderma zelobreve PBMP16, Talaromyces pinophilus PBMP28, Aspergillus flavus KKMP34, and Trichoderma sp. PBMP4 based on their morphological characteristics and multigene (ITS, rpb2, tef-1, CaM, and BenA) phylogenetic analyses. These four endophytic fungi could promote plant growth parameters under greenhouse conditions. Outstandingly, upland rice inoculated with Tr. zelobreve PBMP16 had a significant increase in total seed weight, root length, phenolic compounds, anthocyanin, antioxidants, and N uptake, which were higher than those of the noninoculated control, and even better than the chemical fertilizer. Overall, this report shows that endophytic fungi efficiently promoted growth and increased the phenolic compounds, anthocyanin, and antioxidants of Maled Phai rice.

The isolation of rumen enterococci strains along with high potential utilizing cyanide.

Cyanogenic glycosides in forage species and the possibility of cyanide (CN) poisoning can have undesirable effects on ruminants. The literature estimates that unknown rumen bacteria with rhodanese activity are key factors in the animal detoxification of cyanogenic glycosides, as they are capable of transforming CN into the less toxic thiocyanate. Therefore, identifying these bacteria will enhance our understanding of how to improve animal health with this natural CN detoxification process. In this study, a rhodanese activity screening assay revealed 6 of 44 candidate rumen bacterial strains isolated from domestic buffalo, dairy cattle, and beef cattle, each with a different colony morphology. These strains were identified as belonging to the species Enterococcus faecium and E. gallinarum by 16S ribosomal DNA sequence analysis. A CN-thiocyanate transformation assay showed that the thiocyanate formation capacity of the strains after a 12 h incubation ranged from 4.42 to 25.49 mg hydrogen CN equivalent/L. In addition, thiocyanate degradation resulted in the production of ammonia nitrogen and acetic acid in different strains. This study showed that certain strains of enterococci substantially contribute to CN metabolism in ruminants. Our results may serve as a starting point for research aimed at improving ruminant production systems in relation to CN metabolism.

Promoting growth and production of sunchoke (Helianthus tuberosus) by co-inoculation with phosphate solubilizing bacteria and arbuscular mycorrhizal fungi under drought.

Due to different functions of phosphate solubilizing bacteria (PSB) and arbuscular mycorrhizal fungi (AMF), their potential synergistic effects on enhancing plant growth and yield are worth investigating, especially under adverse conditions. This work focused on the isolation of PSB and characterization for their plant growth promoting properties under drought. The most efficient P solubilizing bacterium was isolated and identified as Burkholderia vietnamiensis strain KKUT8-1. Then, a factorial experiment on the performance of sunchoke (Helianthus tuberosus) was set up with four factors, viz., PSB (presence or absence of KKUT8-1), AMF (presence or absence of Rhizophagus aggregatus), rock phosphate (RP; added or not) and moisture (well-watered (WW) or drought (DS) conditions). Sunchoke performance was enhanced by the presence of AMF, whereas addition of PSB had a positive effect on SPAD values and inulin concentration. Drought reduced plant performance, while addition of RP reduced photosynthetic rate. There was little evidence for synergistic effects between PSB and AMF, except for SPAD values and inulin concentration. Plants that were co-inoculated with AMF and PSB had highest SPAD value, shoot diameter, leaf area, leaf number, chlorophyll concentration, plant biomass, tuber production, root growth and total soluble sugar concentration. Co-inoculated plants also had increased plant water status, reduced electrolyte leakage, and reduced malondialdehyde and proline concentration. Strain KKUT8-1 is the first strain of B. vietnamiensis capable of promoting growth and yield of sunchoke. Enhanced production of sunchoke by a combination of AMF and PSB was much better than the application of RP. Our finding offers an opportunity to develop combinations of biological inoculants for increasing the growth and production of sunchoke under drought in the future.

Enhancement of growth and Cannabinoids content of hemp (Cannabis sativa) using arbuscular mycorrhizal fungi.

This study aimed to investigate the efficiency of arbuscular mycorrhizal fungi (AMF) to promote growth and cannabinoid yield of Cannabis sativa KKU05. A completely randomized design (CRD) was conducted with six replications for 60 days. Two different species of AMF, Rhizophagus prolifer PC2-2 and R. aggregatus BM-3 g3 were selected as inocula and compared with two non-mycorrhizal controls, one without synthetic fertilizer and one with synthetic NPK fertilizer. The unfertilized non-mycorrhizal plants had the lowest performance, whereas plants inoculated with R. aggregatus BM-3 g3 performed best, both in terms of plant biomass and concentrations of CBD and THC. There were no significant differences in plant biomass and cannabinoid concentrations between non-mycorrhizal plants that received synthetic fertilizer and mycorrhizal plants with inoculum of R. prolifer PC2-2. Our data demonstrate the great potential for cannabis cultivation without risking deterioration of soil structure, such as soil hardening and increased acidity, which might be induced by long-term use of synthetic fertilizer.

Bio-strengthening of cementitious composites from incinerated sugarcane filter cake by a calcifying bacterium Lysinibacillus sp. WH.

This study investigated Microbially Induced Calcite Precipitation (MICP) technology to improve the mechanical properties of cementitious composites containing incinerated sugarcane filter cake (IFC) using a calcifying bacterium Lysinibacillus sp. WH. Both IFC obtained after the first and second clarification processes, referred to as white (IWFC) and black (IBFC), were experimented. This is the first work to investigate the use of IBFC as a cement replacement. According to the X-ray fluorescence (XRF) results, the main element of IWFC and IBFC was CaO (91.52%) and SiO2 (58.80%), respectively. This is also the first work to investigate the use of IBFC as a cement replacement. We found that the addition of strain WH could further enhance the strength of both cementitious composites up to ~ 31%, while reduced water absorption and void. Microstructures of the composites were visualized using a scanning electron microscope (SEM). The cement hydration products were determined using X-ray diffraction (XRD) followed by Rietveld analysis. The results indicated that biogenic CaCO3 was the main composition in enhancing strength of the IBFC composite, whereas induce tricalcium silicate (C3S) formation promoting the strength of IWFC composite. This work provided strong evidence that the mechanical properties of the cementitious composites could be significantly improved through the application of MICP. In fact, the strength of IFC-based cementitious composites after boosting by strain WH is only 10% smaller than that of the conventional Portland cement. While using IFC as a cement substitute is a greener way to produce environmentally friendly materials, it also provides a solution to long-term agro-industrial waste pollution problems.

Kinetic model of a newly-isolated Lysinibacillus sp. strain YL and elastic properties of its biogenic CaCO3 towards biocement application.

BACKGROUND: Biocement, calcifying bacteria-incorporated cement, offers an environmentally-friendly way to increase the cement lifespan. This work aimed to investigate the potential use of Lysinibacillus sp. strain YL towards biocement application in both theoretical and experimental ways. METHODS AND RESULTS: Strain YL was grown using calcium acetate (Ca(C2 H3 O2 )2 ), calcium chloride (CaCl2 ) and calcium nitrate (Ca(NO3 )2 ). Maximum bacterial growth of ~0.09 hr-1 and the highest amount of CaCO3 precipitation of ~8.0 g/L were obtained when using Ca(C2 H3 O2 )2 . The SEM and XRD results confirmed that biogenic CaCO3 were calcites. The bulk, Young's and shear moduli of biogenic CaCO3 calculated via the VRH approximation were ~1.5-2.3 times larger than those of ordinary Portland cement. The Poisson's ratio was 0.382 and negative in some directions, suggesting its ductility and auxetic behaviors. The new model was developed to explain the growth kinetic of strain YL in the presence of Ca(C2 H3 O2 )2 , whose concentration was optimized for biocement experiments. Strain YL could increase the compressive strength of cement up to ~50% higher than that of the uninoculated cement. CONCLUSION: Strain YL is a promising candidate for biocement applications. This work represents the trials of experiments and models allowing quantitatively comparison with large-scale production in the future.

Co-Inoculation of an Endophytic and Arbuscular Mycorrhizal Fungus Improve Growth and Yield of Helianthus tuberosus L. under Field Condition.

Endophytic fungi (EPF) and arbuscular mycorrhizal fungi (AMF) symbioses can promote the growth and productivity of several types of plants. This work aimed to investigate the effect of co-inoculation of an EPF Exserohilum rostratum NMS1.5 and an AMF Glomus etunicatum UDCN52867 g.5 on the growth and yields of sunchoke (Helianthus tuberosus L.) compared to the effects of full-dose and half-dose chemical fertilizer (15-15-15) under field conditions. Several plant growth parameters of the co-inoculated plants were significantly higher than the other treatments. Remarkably, such an effect was relatively equal to that of the full-dose chemical fertilizers. Moreover, the co-inoculation of EPF and AMF significantly improved the tuber yield production, even better than the use of a chemical fertilizer. This is the first report to show that plant growth promoting effects of the co-inoculation of EPF and AMF were exceptionally greater than those of the chemical fertilizer. Therefore, our EPF and AMF could potentially be used as a biofertilizer for promoting the growth and yield of sunchoke in the fields.

Purification, characterization and partial amino acid sequences of thermo-alkali-stable and mercury ion-tolerant xylanase from Thermomyces dupontii KKU-CLD-E2-3.

We investigated the properties of the low molecular weight thermo-alkali-stable and mercury ion-tolerant xylanase production from Thermomyces dupontii KKU-CLD-E2-3. The xylanase was purified to homogeneity by ammonium sulfate, Sephadex G-100 and DEAE-cellulose column chromatography which resulted 27.92-fold purification specific activity of 56.19 U/mg protein and a recovery yield of 2.01%. The purified xylanase showed a molecular weight of 25 kDa by SDS-PAGE and the partial peptide sequence showed maximum sequence homology to the endo-1,4-ß-xylanase. The optimum temperature and pH for its activity were 80 °C and pH 9.0, respectively. Furthermore, the purified xylanase can maintain more than 75% of the original activity in pH range of 7.0-10.0 after incubation at 4 °C for 24 h, and can still maintain more than 70% of original activity after incubating at 70 °C for 90 min. Our purified xylanase was activated by Cu2+ and Hg2+ up to 277% and 235% of initial activity, respectively but inhibited by Co2+, Ag+ and SDS at a concentration of 5 mM. The Km and Vmax values of beechwood xylan were 3.38 mg/mL and 625 µmol/min/mg, respectively. Furthermore, our xylanase had activity specifically to xylan-containing substrates and hydrolyzed beechwood xylan, and the end products mainly were xylotetraose and xylobiose. The results suggested that our purified xylanase has potential to use for pulp bleaching in the pulp and paper industry.