Impact of a 5-year winter cover crop rotational system on the molecular diversity of arbuscular mycorrhizal fungi colonizing roots of subsequent soybean

The impact of winter cover crops, specifically wheat (Triticum aestivum L.), red clover (Trifolium pratense L.), and rapeseed (Brassica napus L.) or winter fallow, on community composition of arbuscular mycorrhizal fungi (AMF) in subsequent soybean roots was investigated in a 5-year field trial on andosolic soils in Japan. Soybean roots were sampled at full-flowering and analyzed for AMF communities using a partial LSU rDNA region. Phylogenetic analysis detected 22 AMF phylotypes, including eight Glomus, three Gigaspora, two Scutellospora, three Acaulospora, two Rhizophagus, and one of Funneliformis, Diversispora, Paraglomus, and an unknown glomeromycete in the roots. The 5-year rotation of different winter cover crops or winter fallow did not impact the molecular diversity of AMF communities colonizing the roots of subsequent soybean. In all of the rotations, Glomus and Gigaspora phylotypes were common to soybean roots over the 5-year period. Redundancy analysis (RDA) demonstrated that AMF communities in the roots of subsequent soybean were not significantly different among winter cover crop rotations or fallow. However, AMF communities in soybean roots were clearly influenced by rotation year suggesting that climate or other environmental factors were more important than winter cover cropping system management.     

Biochemical safety evaluation of transgenic rice seeds expressing T cell epitopes of Japanese cedar pollen allergens

Transgenic rice seeds, which express a hybrid peptide comprising seven predominant human T cell epitopes (7Crp) derived from Japanese cedar pollen allergens, have been shown to function as an effective edible vaccine for the control of pollen allergen-induced responses. In this study, we characterized biochemical properties of transgenic seeds expressing the 7Crp peptide. The levels of chemical compositions, such as carbohydrate, protein, lipid, amino acid, fatty acid, mineral, and vitamin, were substantially equivalent between transgenic 7Crp and its nontransgenic counterpart seeds. The contents of three major allergenic proteins in transgenic seeds were not enhanced by expression of the 7Crp peptide when compared with those of nontransgenic seeds. The 7Crp peptide expressed in seeds was susceptible to simulated gastric/intestinal fluids. N-Glycosylation was not observed in the 7Crp peptide sequence. These results indicate that transgenic 7Crp seeds are substantially equivalent to nontransgenic parental seeds except for the presence of the 7Crp peptide. Keywords: Food safety assessment; transgenic rice seed; edible vaccine; peptide-based immunotherapy; Japanese cedar pollinosis.     

Evaluation of the genetic structure of sika deer (Cervus nippon) in Japan's Kanto and Tanzawa mountain areas,based on microsatellite markers

The browsing habits of sika deer (Cervus nippon) in Japan have caused serious ecological problems. Appropriate management of sika deer populations requires understanding the different genetic structures of local populations. In the present study, we used 10 microsatellite polymorphisms to explore the genetic structures of sika deer populations (162 individuals) living in the Kanto region. The expected heterozygosity of the Tanzawa mountain range population (Group I) was lower than that of the populations in the Kanto mountain areas (Group II). Our results suggest that moderate gene flow has occurred between the sika deer populations in the Kanto mountain areas (Group II), but not to or from the Tanzawa mountain range population (Group I). Also, genetic structure analysis showed that the Tanzawa population was separated from the other populations. This is probably attributable to a genetic bottleneck that developed in the Tanzawa sika deer population in the 1950s. However, we found that the Tanzawa population has since recovered from the bottleneck situation and now exhibits good genetic diversity. Our results show that it is essential to periodically evaluate the genetic structures of deer populations to develop conservation strategies appropriate to the specific structures of individual populations at any given time.     

Diffusion of chemicals into archaeological waterlogged hardwoods obtained from the Thang Long Imperial Citadel site,Vietnam

This study shows a method to easily develop suitable impregnation procedure of chemicals for preserving the archaeological waterlogged wood (WW) using diffusion constant. The steady-state diffusion coefficients of polyethylene glycol 4000 (PEG4000), trehalose, and keratin through eight archaeological waterlogged wood species were obtained from the total amounts of the chemicals diffused through the wood samples. The diffusion coefficients were strongly affected by the anatomical structures of the wood species and degree of wood deterioration. Within the same species of wood, the highest diffusion rates were measured in the longitudinal direction, followed by those in the radial and tangential directions. The longitudinal diffusion coefficients were 1.5- to 8.1-times higher than the radial diffusion coefficients, which were in turn approximately 1.1- to 5.7-times higher than the tangential diffusion coefficients. The diffusion rates were found to be inversely proportional to the molecular weight of the diffusing chemical. The diffusivity of trehalose through the waterlogged woods was found to be higher than those of keratin and PEG4000. This study provides a potential method to easily estimate chemical diffusion coefficients for dip-diffusion treatments.     

Relationship between crack propagation and the stress intensity factor in cutting parallel to the grain of hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>)

The method of digital image correlation (DIC) was applied to the digital image of orthogonal cutting parallel to the grain of hinoki, and the strain distribution near the cutting edge was evaluated. The wood fracture associated with chip generation was considered as mode I fracture, and the stress intensity factor K_I for fracture mode I was calculated from the strain distribution according to the theory of linear elastic fracture mechanics for the anisotropic material. The calculated K_I increased prior to crack propagation and decreased just after the crack propagation. The change in K_I before and after crack propagation, ΔK_I, decreased in accordance with the crack propagation length, although the variance in ΔK_I should depend on the relationships between the resolution of DIC method and the dimensions of cellular structure. The calculated K_I in this study was almost on the same order as reported in the literatures. It was also revealed, for the case of chip generation Type 0 or I, the stress intensity factor for fracture mode II could be negligible due to the higher longitudinal elastic properties of wood in the tool feed direction than the one radial ones, and the mode I fracture was dominant.     

Trichomes: interaction sites of tomato leaves with biotrophic powdery mildew pathogens

The present study aimed to explore the possibility of using the type I trichomes of tomato (Solanum lycopersicum) to monitor the infection processes of powdery mildews by microscopy. Individual trichome cells of two tomato genotypes were inoculated with pathogenic and non-pathogenic powdery mildew species, Pseudoidium neolycopersici, Erysiphe trifoliorum and Podosphaera xanthii. On the trichome cells of the tomato cultivar Moneymaker, hyphae of the pathogenic Pseudoidium neolycopersici (isolates KTP-03 and KTP-04) grew vigorously; whereas hyphal growth of the non-pathogenic Erysiphe trifoliorum and Podosphaera xanthii ceased after appressorium formation, which was associated with papilla formation and hypersensitive cell death, respectively. Similar infection processes of the tested powdery mildews were seen in Moneymaker epidermal cells. Therefore, tomato trichomes are suitable for analysing, at individual cell level, the infection processes of different pathotypes of powdery mildews and for observing the cytological responses of plants by non-pathogenic powdery mildews. On the other hand, it was observed that both isolates KTP-03 and KTP-04 failed to produce conidiophores on the hyphae elongating on Moneymaker trichomes. Similarly, no conidiophores were produced on the hyphae elongating on trichomes of Solanum peruvianum LA2172, which is resistant to KTP-03 and susceptible to KTP-04. Interestingly, delayed cell death occurred in LA2172 epidermal cells, which were attacked by KTP-03 hyphae elongating from trichomes and conidiophores were formed on new hyphae growing from the leaf epidermal cells. Thus, leaf trichomes and epidermal cells of the wild tomato species LA2172 reacted differently to the avirulent isolate KTP-03.     

Analysis of PPP1R11 expression in granulosa cells during developmental follicles of yak and its effects on cell function

The aims of this study were to analyse the protein phosphatase 1 regulatory subunit 11 (PPP1R11) expression and cellular localization in yak follicles and investigate its effects on cell proliferation, apoptosis and oestrogen secretion in granulosa cells (GCs). Ten healthy and non-pregnant female yaks (4-year-old) were used as experimental animals. The mRNA relative expression level of PPP1R11 in GCs from small (<3.0 mm), medium (3.0–5.9 mm) and large (6.0–9.0 mm) follicles was detected by RT-qPCR, and the cellular localization of PPP1R11 protein was detected by immunohistochemistry staining (IHC). After isolation, culture and identification of yak GCs in vitro, si-PPP1R11 and si-NC (negative control) were transfected into GCs. RT-qPCR and immunofluorescence staining were used to evaluate the interference efficiency, and ELISA was performed to detect oestrogen concentration. Then, EdU staining and TUNEL staining were conducted to analyse cell proliferation and apoptosis. In addition, the oestrogen synthesis, proliferation- and apoptosis-related genes were detected by RT-qPCR after knockdown PPP1R11. The results showed that PPP1R11 is mainly located in ovarian GCs, and the expression levels of PPP1R11 in GCs from large follicles were significantly higher than that from medium and small follicles. Transfection of si-PPP1R11 into GCs could significantly inhibit the expression of PPP1R11. Interestingly, the oestrogen secretion ability and the expression level of oestrogen pathway-related genes (STAR, CYP11A1, CYP19A1 and HSD17B1) were also significantly downregulated. Moreover, the proportion of positive cells was decreased, and cellular proliferation-related genes (PCNA, CCNB1 and CDC25A) were significantly downregulated after knockdown PPP1R11. However, the proportion of apoptotic cells was increased, and apoptosis-related genes (BAX, CASP3 and P53) were significantly upregulated. Taken together, this study was the first revealed the expression and cellular localization of PPP1R11 in yak follicles. Interference PPP1R11 could reduce oestrogen secretion, inhibit proliferation and promote apoptosis in GCs, which provided a basis for further studies on the regulatory mechanism of PPP1R11 in follicle development.