Screening of endophytic bacteria against fungal plant pathogens

Bacterial endophytes were found from 6 plant leaves among 35 plant leaves screened. Two of the isolated bacteria showed antagonistic activity against fungal plant pathogens. An isolate named KL1 showed the clear inihibition against plant pathogens, Fusarium oxysporum and Rhizoctonia solani, on PDA as well as TSA plate. Supernatant of the bacterial culture also showed the clear inhibition against the fungal growth on the plate and the antibiotic substance was identified as iturin A by HPLC analysis. KL1 was identified as Bacillus sp. from the 16S rRNA gene analysis. Very thin hyphae of R. solani was miccroscopically observed when the fungus was co-cultivated with KL1.     

Feasible conditions for Japanese woody biomass utilization

Environmental Science and Pollution Research - There is an abundance of woody biomass in Japan. However, its economic feasibility is limited. There have been several discussions on whether...     

Identifying a breeding habitat of a critically endangered fish, <Emphasis Type="Italic">Acheilognathus typus</Emphasis>, in a natural river in Japan

Freshwater biodiversity has been severely threatened in recent years, and to conserve endangered species, their distribution and breeding habitats need to be clarified. However, identifying breeding sites in a large area is generally difficult. Here, by combining the emerging environmental DNA (eDNA) analysis with subsequent traditional collection surveys, we successfully identified a breeding habitat for the critically endangered freshwater fish Acheilognathus typus in the mainstream of Omono River in Akita Prefecture, Japan, which is one of the original habitats of this species. Based on DNA cytochrome B sequences of A. typus and closely related species, we developed species-specific primers and a probe that were used in real-time PCR for detecting A. typus eDNA. After verifying the specificity and applicability of the primers and probe on water samples from known artificial habitats, eDNA analysis was applied to water samples collected at 99 sites along Omono River. Two of the samples were positive for A. typus eDNA, and thus, small fixed nets and bottle traps were set out to capture adult fish and verify egg deposition in bivalves (the preferred breeding substrate for A. typus) in the corresponding regions. Mature female and male individuals and bivalves containing laid eggs were collected at one of the eDNA-positive sites. This was the first record of adult A. typus in Omono River in 11 years. This study highlights the value of eDNA analysis to guide conventional monitoring surveys and shows that combining both methods can provide important information on breeding sites that is essential for species’ conservation.     

Rate of hexabromocyclododecane decomposition and production of brominated polycyclic aromatic hydrocarbons during combustion in a pilot-scale incinerator

Here, we examined the incineration of extruded polystyrene containing hexabromocyclododecane(HBCD) in a pilot-scale incinerator under various combustion temperatures(800–950℃) and flue gas residence times(2–8 sec). Rates of HBCD decomposition ranged from 99.996%(800℃, 2 sec) to 99.9999%(950℃, 8 sec); the decomposition of HBCD, except during the initial stage of combustion(flue gas residence time 2 sec), followed a pseudo-first-order kinetics model. An Arrhenius plot revealed that the activation energy and frequency factor of the decomposition of HBCD by combustion were 14.2 kJ/mol and 1.69 sec~(-1),respectively. During combustion, 11 brominated polycyclic aromatic hydrocarbons(BrPAHs)were detected as unintentional by-products. Of the 11 BrPAHs detected, 2-bromoanthracene and 1-bromopyrene were detected at the highest concentrations. The mutagenic and carcinogenic BrPAHs 1,5-dibromoanthracene and 1-bromopyrene were most frequently detected in the flue gases analyzed. The total concentration of BrPAHs exponentially increased(range,87.8–2,040,000 ng/m~3) with increasing flue gas residence time. Results from a qualitative analysis using gas chromatography/high-resolution mass spectrometry suggest that bromofluorene and bromopyrene(or fluoranthene) congeners were also produced during the combustion.     

Effect of temperature on growth and maturation pattern of <Emphasis Type="Italic">Caprella mutica</Emphasis> (Crustacea,Amphipoda): does the temperature–size rule function in caprellids?

Many ectothermal marine animals mature at larger sizes in lower temperature environments and at smaller sizes in higher temperature environments. This phenomenon is called the temperature–size rule. To examine whether this rule controls the appearance of large adults in a winter population of caprellids, individuals of Caprella mutica were reared at different temperatures. Caprellids at 5°C died at instar III before they reached maturity. In contrast, the animals reared at 10, 15 and 20°C lived to higher instars and reached maturity within their lifetime. Somatic growth pattern did not change between 10 and 20°C. Maturation instar of males was not affected by temperature. This indicates that the appearance of large adults in winter is not a result of a change in somatic growth pattern with temperature change. However, female maturation size becomes larger due to a delay in maturation at lower temperature. This, in turn, indicates that the temperature–size rule plays a role in the mechanism controlling the appearance of large female adults in winter populations.     

Differences of two polychaete species reflected in enzyme activities

Polychaetes constitute most of the benthic macroinvertebrates in estuarine and coastal environments. We investigated the utilization of organic matter in two polychaete species, Capitella sp. I and Perinereis nuntia brevicirris, living in different coastal habitats. The protease activity of Capitella sp. I (89.7 μg mg⁻¹) was about 10 times that of P. nuntia brevicirris (8.0 μg mg⁻¹). High cellulase (endo-β-1,4-glucanase) activity was detected in P. nuntia brevicirris (3.2 μg mg⁻¹), whereas scarcely any was detected in Capitella sp. I. We isolated cDNA clones of protease mRNA from Capitella sp. I and of cellulase mRNA from P. nuntia brevicirris. The high protease activity of Capitella sp. I enabled it to survive in the sediment under a fish farm, where it degrades organic matter. In contrast, the high cellulase activity of the estuary-dwelling P. nuntia brevicirris allowed it to degrade organic matter originating from terrestrial areas.     

Morphological and habitat divergence in the intertidal limpet Patelloida pygmaea

In the intertidal limpet Patelloida pygmaea, two distinctive morphs, the forms pygmaea and conulus, have been recorded. The former possesses a flat elongated shell, and the latter has an extremely high round shell. It has been observed in the field that pygmaea is found on oyster shells Crassostrea gigas. The form conulus uses an unusual substrate for attachment. It is found on the living shells of the intertidal gastropod Batillaria cumingi. Although conulus is normally found only on shells of Batillaria, it can also be found on oyster shells when pygmaea and Batillaria shells are not present in nature. An electrophoretic analysis of allozymes showed that these two forms are reproductively isolated from each other and coexist without gene exchange on the same mudflat. Laboratory experiments showed that pygmaea prefers oyster shells and conulus prefers Batillaria shells as substrates for attachment when both oyster and Batillaria shells are present. The form pygmaea did not attach to Batillaria shells, even when only Batillaria shells were available. However, conulus also attached to oyster shells when Batillaria shells were not available. The proportion of individuals of conulus that attached to oyster shells decreased significantly when pygmaea was attaching to the oyster shells. These results suggest that pygmaea is ecologically more specialized to living on oyster shells than conulus.     

Production characteristics of lipopeptide antibiotics in biofilm fermentation of Bacillus subtilis

In biofilm fermentation, from the very early moments, surfactin was produced along with the biofilm development in the lipopeptide antibiotic production medium by using Bacillus subtilis. However, almost no iturin A was produced in its first 24 hours of cultivation and the production of iturin A began much later. Volumes of the nutrient medium and available surface area of the biofilm reactors were found to be important with the relative production of these two antibiotics. Production of iturin A was increased from 12 mg to about 50 mg per reactor when the culture size was increased from 5 mL to 20 mL, as the depth of the medium was increased. The production level was saturated thereafter with larger volumes. On the other hand, surfactin production was remained similar, which was about 10 mg per reactor, from all the 5 mL to 80 mL of biofilm culture. Optimized temperature for iturin A and surfactin production was observed at 25 and 37°C, respectively. In the biofilm fermentation, production of surfactin was increased when the incubation temperature was increased within the temperature range of 25 to 37°C, on the other hand, iturin A production was gradually decreased with the increase of the incubation temperature.