Developing Actinobacterial Endophytes as Biocontrol Products for Fusarium pseudograminearum in Wheat

Crown rot of wheat, caused by Fusarium pseudograminearum, results in millions of dollars of yield losses globally each year. Management strategies to control crown rot are limited and there are concerns about development of fungicide resistance so novel treatment strategies are desirable. A collection of endophytic Actinobacteria was screened for their ability to suppress the growth of F. pseudograminearum and the development of crown rot symptoms in wheat with the aim of identifying candidates that can be developed into biocontrol products. The ability of the Actinobacteria isolates to suppress the growth of three different F. pseudograminearum strains in vitro was assessed using agar-plate competition assays. Soil-free seedling assays were used to screen for suppression of development of early disease symptoms in the susceptible wheat (Triticum aestivum) cv. Tamaroi. Four of the isolates were tested in a glasshouse pot experiment to assess their ability to decrease disease symptoms and prevent yield losses in wheat cv. Tamaroi grown to maturity in an unsterilized soil. The screening of 53 isolates identified two Streptomyces isolates, MH71 and MH243, with very strong antifungal activity against F. pseudograminearum strains in agar-plate competition and seedling assays. In the glasshouse pot trial, plants treated with seed coatings of either MH71 or MH243 had > 24% lower disease severity than control plants infected with F. pseudograminearum. These two cultures show potential for development as biocontrol products because they are easy to culture, grow on relatively inexpensive media, produce highly durable spores and can be delivered to plants as a seed coat.

Soil microbes and oxygen influence the changes in bacterial community composition during swine carcass decomposition

Background Carcass decomposition is influenced by various factors such as temperature, humidity, microorganisms, invertebrates, and scavengers. Soil microbes play a significant role in the decomposition process. In this study, we investigated the changes in the bacterial community during carcass decomposition in soil with an intact microbial community and soil which was sterilized decomposed with and without oxygen access using 16s rRNA metagenomic sequencing. Results Based on the 16S rRNA metagenomic sequencing, a total of 988 operational taxonomic units (OTUs) representing 16 phyla and 533 genera were detected. The bacterial diversity varied across the based on the alpha diversity indices. The bacterial composition in the unsterilized soil – aerobic condition (U_A) and unsterilized soil – anaerobic condition (U_An) set-ups have higher alpha diversity than the other burial set-ups. Beta diversity analysis revealed a close association in the samples according to the burial type and decomposition day. Firmicutes was the dominant phylum across all samples regardless of the burial type and decomposition day. The bacterial community composition changed throughout the decomposition process in all burial set-up. Meanwhile, the genus Bacillus dominated the bacterial community towards the end of decomposition period. Conclusions Our results showed that bacterial community composition changed during carcass decomposition and was affected by the soil and oxygen access, with microorganisms belonging to phylum Firmicutes dominating the community.

Comparative effect of Rhizophagus irregularis strain on cassava root development and Phosphorus uptake under acidic soils conditions of Walungu territory, Eastern DR Congo.

Objective: In the highlands of South-Kivu province of DR Congo, cassava is grown on marginal land not suitable for other crops. Walungu territory for instance is dominated by acidic soils and has the highest nutrient depletion rate in the country. On such types of soil, nutrient depletion is accompanied by a decrease in the availability of phosphorus and many other nutrients. The use of Arbuscular mycorrhizal fungi (AMF) would therefore be an alternative to improve the phosphate nutrition of cassava under different soil conditions. Methodology and results: The experiment reported in the current study was conducted in pots under semi-controlled conditions. Ferrallitic soil from Walungu was used. Soil was sterilized or not and then inoculated with the AMF Rhizophagus irregularis (RI) strain. Results showed that inoculation with an exotic AMF strain (Rhizophagus iregularis) has significantly increased root abundance (number of roots) in both sterilized and unsterilized soil and root dry weight in sterilized soil only. However, in unsterilized soil, root dry weight decreased with AMF inoculation. In Walungu acidic soils, P supply could significantly influence the effect of mycorrhizal inoculation on root development and root abundance. AMF inoculation has significantly increased the shoot P concentration when P was supplied. In sterilized soil, mycorrhizal inoculation had a depressant effect on soil phosphorus concentration, especially when P was not supplied. Conclusion and application of results: Our results suggest that the introduced Rhizophagus iregularis strain increases P uptake in the rhizospheric soil, especially when phosphorus is not applied to the sterilized or unsterilized soil. The use of the Rhizophagus irregularis strain as a biofertilizer could improve phosphorus nutrition and root development in cassava. Keywords: Arbuscular mycorrhizae, Rhizophagus irregularis, P uptake, Cassava, Ferrallitic soil

Arbuscular Mycorrhizal Fungi Can Compensate for the Loss of Indigenous Microbial Communities to Support the Growth of Liquorice (Glycyrrhiza uralensis Fisch.)

Soil microorganisms play important roles in nutrient mobilization and uptake of mineral nutrition in plants. Agricultural management, such as soil sterilization, can have adverse effects on plant growth because of the elimination of indigenous microorganisms. Arbuscular mycorrhizal (AM) fungi are one of the most important beneficial soil microorganisms for plant growth. However, whether AM fungi can compensate for the loss of indigenous microbial communities to support plant growth and metabolism is largely unknown. In this study, a pot experiment was conducted to investigate the effects of AM fungi on plant growth and secondary metabolism in sterilized and unsterilized soil. We used liquorice (Glycyrrhiza uralensis Fisch.), an important medicinal plant as the host, which was inoculated with the AM fungus Rhizophagus irregularis or not and grown in unsterilized or sterilized soil. Plant photosynthesis traits, plant growth and nutrition level, concentrations of the secondary metabolites, and expression levels of biosynthesis genes were determined. The results showed that soil sterilization decreased plant growth, photosynthesis, and glycyrrhizin and liquiritin accumulation, and moreover, downregulated the expression of related biosynthesis genes. Inoculation with R. irregularis in sterilized soil offset the loss of indigenous microbial communities, resulting in plant growth and glycyrrhizin and liquiritin concentrations similar to those of plants grown in unsterilized soil. Thus, AM fungi could compensate for the loss of indigenous microbial communities by soil sterilization to support plant growth and secondary metabolism.

Integrated Effects of Co-Inoculation with Phosphate-Solubilizing Bacteria and N2-Fixing Bacteria on Microbial Population and Soil Amendment Under C Deficiency

The inoculation of beneficial microorganisms to improve plant growth and soil properties is a promising strategy in the soil amendment. However, the effects of co-inoculation with phosphate-solubilizing bacteria (PSB) and N2-fixing bacteria (NFB) on the soil properties of typical C-deficient soil remain unclear. Based on a controlled experiment and a pot experiment, we examined the effects of PSB (M: Bacillus megaterium and F: Pseudomonas fluorescens), NFB (C: Azotobacter chroococcum and B: Azospirillum brasilence), and combined PSB and NFB treatments on C, N, P availability, and enzyme activities in sterilized soil, as well as the growth of Cyclocarya Paliurus seedlings grow in unsterilized soil. During a 60-day culture, prominent increases in soil inorganic N and available P contents were detected after bacteria additions. Three patterns were observed for different additions according to the dynamic bacterial growth. Synergistic effects between NFB and PSB were obvious, co-inoculations with NFB enhanced the accumulation of available P. However, decreases in soil available P and N were observed on the 60th day, which was induced by the decreases in bacterial quantities under C deficiency. Besides, co-inoculations with PSB and NFB resulted in greater performance in plant growth promotion. Aimed at amending soil with a C supply shortage, combined PSB and NFB treatments are more appropriate for practical fertilization at intervals of 30–45 days. The results demonstrate that co-inoculations could have synergistic interactions during culture and application, which may help with understanding the possible mechanism of soil amendment driven by microorganisms under C deficiency, thereby providing an alternative option for amending such soil.

Repellent Effects of Methyl Anthranilate on Western Corn Rootworm Larvae (Coleoptera: Chrysomelidae) in Soil Bioassays

Methyl anthranilate (MA), a compound in maize roots that is repellent to western corn rootworm larvae (Diabrotica virgifera virgifera LeConte) was tested in behavioral bioassays in a soil environment. MA prevented larvae from locating roots of a maize seedling, and the repellency strengthened with increasing rates of MA. In a simple push–pull strategy between an MA-treated seedling and an untreated seedling, granules containing 0.1 mg/g MA pushed larvae to the untreated seedling. This push effect increased with dose, with 90% repellency observed for the highest dose tested (100 mg/g). Chemical analysis showed that MA concentrations remained high for 4 wk in dry, sterilized or unsterilized soil, but declined rapidly in moist soil. After 7 d, 50% less MA was recovered in moist, sterilized soil than in dry soil, and only a trace of MA remained in unsterilized moist soil, suggesting that both moisture and microbial activity contributed to the loss of MA. Various (MA) carrier granules were tested in bioassays after aging in moist soil. After 1 d, all of the MA granules were repellent at the 10 mg/g rate and clay granules were also effective at 1 mg/g. After 1 wk, only molecular sieve granules elicited repellency, but that activity disappeared after 2 wk. These results demonstrate that MA is repellent to western corn rootworm larvae in the soil environment and may have potential as a rootworm treatment if formulations can be developed that protect the material from decomposition in the soil.

BIOCONTROL OF FUSARIUM OXYSPORUM IN UNSTERILIZED SOIL BY NOVEL STREPTOMYCES CACAOI SUBSP CACAOI [M20]

Objective: To find bio fungicide from mangrove actinomycetes for controlling seed and soil borne pathogen-Fusarium oxysporum. Methods: A total of 25 actinomycetes were isolated by pour plate method. These were screened for fungicidal activity by agar plug method. The isolate M20 was characterised further for identification. The phytotoxicity study was done; biocontrol of Fusarium oxysporum with 10% culture filtrate was done using food poisoning technique. Volatile toxicity of isolate M20 was studied by inverted plate technique. The methanolic crude extract was subjected for UV–Vis spectral analysis for identifying the group of compound present.Results: The isolated M20 found to be better in antifungal activity. 10% culture filtrate actively inhibited the growth of Fusarium oxysporum (77.7%), 10% culture filtrate was taken as a standard concentration for biocontrol of Fusarium oxysporum using green gram as the test plants. The 15th-day green gram plants under treatment with the antagonist (A), antagonist+pathogen (A+P), antagonist+pathogen+rhizobium (A+P+R) yielded high biomass and better growth. The disease development by the pathogen in green gram was controlled by the antagonist. The compounds (pyrimidine nucleosides-neutral and acidic polyoxins (230 nm), (270-290 nm) and heptaene antifungal antibiotics (406-417 nm)) are preliminarily confirmed from the methanolic crude extract of the isolate M20-Streptomyces cacaoi subsp cacaoi.Conclusion: Since the isolate M20 controlled the growth and disease causing potentiality of Fusarium oxysporum, it can be effectively used to control seed and soil borne diseases that are caused by Fusarium oxysporum.