Effect of winter wheat cover cropping with no-till cultivation on the community structure of arbuscular mycorrhizal fungi colonizing the subsequent soybean

Winter cover crops increase the amount of indigenous arbuscular mycorrhizal fungi (AMF) in the soil, providing beneficial effects such as enhancement of phosphorus uptake by the subsequent crop. However, its impact on the AMF community structure is not well understood. In the present study, we aimed to reveal the effect of winter wheat cover cropping with no-till cultivation on the AMF community structures in soil and roots of the subsequent soybean. For this purpose, we conducted a field experiment consisting of two treatments, no-till soybean cultivation after winter wheat cover cropping (NTWC) and conventional soybean cultivation after winter fallow management as a control (CONT). At the flowering stage of soybean, higher AMF colonization of soybean roots was observed in the NTWC plots compared with the CONT plots. Additionally, aboveground biomass and phosphorus uptake of soybean in the NTWC plots were significantly higher than those in the CONT plots. Molecular community analyses based on PCR-denaturing gradient gel electrophoresis (DGGE) of AMF 18S rRNA genes indicated that the AMF community structures in the soil and soybean root of the NTWC plots were clearly different from those of the CONT plots. The DGGE profiles showed that the wheat cover cropping preferentially increased some phylotypes belonging to Glomeraceae and Claroideoglomeraceae. In addition, most of the phylotypes were characteristically observed in the subsequent soybean root of the NTWC plots, strongly suggesting that these phylotypes colonizing the cover crop wheat were taken over by the subsequent soybean. Our study revealed the significant effect of winter cover cropping with no-till cultivation on the structure of AMF community colonizing the subsequent soybean.     

Characterization of bagasse binderless particleboard manufactured in high-temperature range

This paper describes the features of binderless particleboard manufactured from sugarcane bagasse, under a high pressing temperature of 200–280 °C. Mechanical properties [i.e., modulus of rupture (MOR) and elasticity (MOE) in dry and wet conditions, internal bonding strength (IB)] and dimensional stability [i.e., thickness swelling (TS)] of the board were evaluated to investigate the effect of high pressing temperature. Recycled chip binderless particleboards were manufactured under the same conditions for comparison, and particleboards bonded with polymeric methylene diphenyl diisocyanate (PMDI) resin were manufactured as reference material. The target density was 0.8 g/cm³ for all of the boards. The results showed that the mechanical properties and dimensional stability of both types of binderless boards were improved by increasing the pressing temperature. Bagasse showed better performance than that of recycled chip as a raw material in all evaluations. Bagasse binderless particleboard manufactured at 260 °C had an MOE value of 3.5 GPa, which was equivalent to the PMDI particleboard, and a lower TS value of 3.7 % than that of PMDI particleboard. The MOR retention ratio under the dry and wet conditions was 87.0 %, while the ratio for the PMDI particleboard was only 54.6 %. The obtained results showed the possibility of manufacturing high-durability binderless particleboard, with good dimensional stability and water resistance, which previously were points of weakness for binderless boards. Manufacturing binderless boards under high temperature was effective even when using particles with poor contact area, and it was possible to express acceptable properties to allow the manufacture of particleboards. Further chemical analysis indicated a contribution of a saccharide in the bagasse to the improvement of the board properties.     

A single gene transfer of gibberellin biosynthesis gene cluster increases gibberellin production in a Fusarium fujikuroi strain with gibberellin low producibility

Fusarium fujikuroi, the causative agent of bakanae disease in rice, produces many kinds of secondary metabolites. Recently, two phylogenetic subgroups (F and G groups) of Japanese F. fujikuroi have been identified and found to have differences in their gibberellin (GA) and fumonisin production. G-group F. fujikuroi produces large amounts of GA, but is a fumonisin nonproducer. F-group produces large amounts of fumonisin, but is a GA low or nonproducer. We investigated the cause of low GA production in the F-group. Genetic mapping suggests that low GA production in the F-group strain Gfc0825009 is due to a GA gene cluster for GA biosynthesis. Analysis of the nucleotide and amino acid sequences of the genes in the GA gene cluster showed >98.4% homology between the F-group strain Gfc0825009 and the G-group strain Gfc0801001. Following a 7-day culture under low nitrogen conditions, we found that expression of P450-1, P450-4, and P450-2 in the cluster increased in the G-group strain and not in the F-group strain. We hypothesized that complementation by GA genes in the G-group strain would be required to increase GA production in the F-group strain. However, we found that this occurred with a single gene complementation of DES, P450-1, P450-4, or P450-2. Simultaneous increase in the expression of P450-1, P450-4, and P450-2 were detected in the complementary transformants. Moreover, the same phenomenon was observed by reintegration of its own P450-1. Our results suggest the presence of unknown regulatory mechanisms of the GA gene cluster in F. fujikuroi.     

The effects of puroindoline b on the ultrastructure of endosperm cells and physicochemical properties of transgenic rice plant

Endosperm texture is an important factor governing the end-product quality of cereals. The texture of wheat (Triticum aestivum L.) endosperm is controlled by puroindoline a and b genes which are both absent in rice (Oryza sativa L.). It has been reported that the endosperm texture of rice can be modified by puroindoline genes. The mechanism, however, by which puroindolines affect the ultrastructure of rice endosperm cells remains to be investigated. In this study, we observed the ultrastructure of endosperm cells and the morphology of isolated starch granules of the transgenic rice expressing the puroindoline b gene. SEM and TEM observations indicated that compound starch granules were embedded within the matrix material in non-transgenic rice, Nipponbare, whereas they were surrounded by spaces in the transgenic rice. The morphology and size of each starch granule were not different between non-transgenic and the transgenic rice. However, the transgenic rice flour showed smaller particle size, higher starch damage, and lower viscosity during gelatinization than that of non-transgenic rice. These results confirm that puroindoline b reduces the grain hardness in rice. Moreover, the results also suggest that puroindoline b functions at the surface of compound starch granules, and not on polygonal starch granules in rice endosperm.     

Shading of net cage is an effective control measure against skin fluke Neobenedenia girellae infection in chub mackerel Scomber japonicus

The skin fluke Neobenedenia girellae has become a serious problem in Japan since the 1990s. Present control methods focus on the removal of the attached parasite and these post-infection treatments are often labor intensive, time consuming, and/or stressful to fish. Chub mackerel Scomber japonicus are highly susceptible to N. girellae. However, because of their sensitive nature, bath treatments may cause mortality. In this study, we investigated the efficacy of cage shading to reduce skin fluke infection and the frequency of conventional post-infection treatments. Juvenile mackerel were reared in cages with or without shade for 3 months and their skin fluke infections were monitored. We performed either freshwater baths or oral administration of praziquantel if fluke intensity exceeded the given criteria. In unshaded cages, 3 total bath treatments or 6 total drug treatments were conducted. In contrast, no treatment was required for the shaded cage. The overall fluke intensity in the shaded cage was less than half that of the unshaded cages, despite the lack of treatments. This study demonstrated for the first time the practical use of shading in fish farms to reduce skin fluke infection.     

Seedling blight of Glycyrrhiza uralensis caused by Pythium myriotylum,P. aphanidermatum and P. spinosum and identifying primary inoculum sources using multiplex PCR detection

Pythium species, isolated from seedlings of Glycyrrhiza uralensis with blight, were identified as P. myriotylum, P. aphanidermatum, and P. spinosum on the basis of morphological characteristics and sequences of the internal transcribed spacer regions of rDNA. In pathogenicity tests, the isolates of the three Pythium species caused blight, producing the original disease symptoms. The primary inoculum source was determined using a multiplex PCR to detect the pathogen. All the Pythium species were detected in the soils of fields with the diseased plants and in soils of adjacent field soils.     

Structure and activity of soil-inhabiting methanotrophic communities in northern forest of Japan

In a Japanese forest, CH₄ uptake rate and methanotrophic community structure in the soil were investigated at four sites of different vegetation. At two of these sites, undergrowth was dominated by Sasa senanensis, and that of another was dominated by Sasa kurilensis. At the rest site, undergrowth had been removed artificially. The tree-layer composition differed between the two sites with S. senanensis, but tree layer of the other two sites were dominated by the same species. At the site lacking undergrowth, observed CH₄ uptake rate was twice as high as that at the other sites. Under laboratory conditions, soil sample from the site lacking undergrowth exhibited CH₄ consumption rate higher than that of the adjacent site with the same dominating tree species. The community structures of methanotrophs were investigated with denaturing gradient gel electrophoresis (DGGE) of the gene encoding particulate methane monooxygenase (pmoA). The banding patterns observed were different depending on the type of undergrowth vegetation. The sequences of the DGGE bands were closely related to each other and belonged to the “upland soil cluster alpha” (USCα). These results imply possible close relationship between the undergrowth vegetation and methanotrophic communities in forest soils.     

PCR primers for identification of opine types of <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> in Japan

The polymerase chain reaction (PCR) is a rapid, precise method for detecting and identifying pathogenic bacteria. In addition to the published primers for identification of Agrobacterium tumefaciens up to species level, two sets of primers were designed to identify the nopaline and octopine types of Agrobacterium tumefaciens. The RBF-RBR primer set designed based on the nopaline type T-DNA right border detected the nopaline type A208 and R225f strains, and the ocsF-ocsR primer set derived from the ocs gene of the octopine type A. tumefaciens detected the octopine type A348 strain. After polymerase Chain reaction (PCR) amplification by the RBF-RBR primers, the A208 and R225f strains could be differentiated from each other by restriction fragment length polymorphism digestion using the restriction enzymes DraI and XbaI. Multiple colonies can be screened at one time in a single PCR tube with satisfactory efficiency, thereby allowing rapid detection of pathogenic A. tumefaciens. Following a rough screening by classical biovar medium and -methyl-d-glucoside medium, the developed PCR system was introduced to identify isolates collected from soil and crown gall samples. Of 42 isolates determined to be A. tumefaciens, 7 were found to be octopine type; all the rest were R225f type.     

Comparisons of single versus multiple bacterial species on biological control of anthurium blight

ABSTRACT Effects of single versus multiple biological control agents (BCAs) on suppression of bacterial blight of anthurium were studied using a bioluminescent strain (V108LRUH1) of Xanthomonas campestris pv. dieffenbachiae. When five BCAs (GUT3, GUT4, GUT5, GUT6, and GUT9) were coinoculated in various combinations with V108LRUH1 into filter-sterilized guttation fluids of anthurium plants, a mixture of all five strains or four strains without GUT9 was most inhibitory to V108LRUH1. None of the individual BCAs inhibited V108LRUH1 in the guttation fluid. When BCAs were sprayed at congruent with10(8) CFU/ml on the foliage of a susceptible cultivar 1 day prior to inoculation with V108LRUH1, GUT6 alone and any mixtures containing GUT6 were highly effective in suppressing wound invasion and subsequent leaf infection by V108LRUH1. When tested on several cultivars that differed in susceptibility to the disease, the mixture of five strains or four strains without GUT9 consistently suppressed leaf infection regardless of the cultivars. In some cultivars, BCAs completely suppressed both wound and hydathode invasion by V108LRUH1, resulting in no infection in many leaves. These results indicate that application of bacterial mixtures provides anthurium cultivars with bacterial communities suppressive to X. campestris pv. dieffenbachiae. The results also suggest that selecting an effective mixture of BCAs first and then removing ineffective strains may be a better general approach to finding the most effective BCAs than finding individual strains and combining them.