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31.
A typhoon event catastrophically destroyed a 45-year-old Japanese larch plantation in southern Hokkaido, northern Japan in September 2004, and about 90% of trees were blown down. Vegetation was measured to investigate its regeneration process and CO2 flux, or net ecosystem production (NEP), was measured in 2006–2008 using an automated chamber system to investigate the effects of typhoon disturbance on the ecosystem carbon balance. Annual maximum aboveground biomass (AGB) increased from 2.7 Mg ha−1 in 2006 to 4.0 Mg ha−1 in 2007, whereas no change occurred in annual maximum leaf area index (LAI), which was 3.7 m2 m−2 in 2006 and 3.9 m2 m−2 in 2007. Red raspberry (Rubus idaeus) had become dominant within 2 years after the typhoon disturbance, and came to account for about 60% and 50% of AGB and LAI, respectively. In comparison with CO2 fluxes measured by the eddy covariance technique in 2001–2003, for 4.5 months during the growing season, the sum of gross primary production (GPP) decreased on average by 739 gC m−2 (64%) after the disturbance, whereas ecosystem respiration (RE) decreased by 501 gC m−2 (51%). As a result, NEP decreased from 159 ± 57 gC m−2 to −80 ± 30 gC m−2, which shows that the ecosystem shifted from a carbon sink to a source. Seasonal variation in RE was strongly correlated to soil temperature. The interannual variation in the seasonal trend of RE was small. Light-saturated GPP (Pmax) decreased from 30–45 μmol m−2 s−1 to 8–12 μmol m−2 s−1 during the summer season through the disturbance because of large reduction in LAI.  相似文献   
32.
Epigenetic alteration is an emerging paradigm underlying the long-term effects of chemicals on gene functions. Various chemicals, including organophosphate insecticides and heavy metals, have been detected in the human fetal environment. Epigenetics by DNA methylation and histone modifications, through dynamic chromatin remodeling, is a mechanism for genome stability and gene functions. To investigate whether such environmental chemicals may cause epigenetic alterations, we studied the effects of selected chemicals on morphological changes in heterochromatin and DNA methylation status in mouse ES cells (ESCs). Twenty-five chemicals, including organophosphate insecticides, heavy metals and their metabolites, were assessed for their effect on the epigenetic status of mouse ESCs by monitoring heterochromatin stained with 4¢,6-diamino-2-phenylindole (DAPI). The cells were surveyed after 48 or 96 h of exposure to the chemicals at the serum concentrations of cord blood. The candidates for epigenetic mutagens were examined for the effect on DNA methylation at genic regions. Of the 25 chemicals, five chemicals (diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se) and octachlorodipropyl ether (S-421)) caused alterations in nuclear staining, suggesting that they affected heterochromatin conditions. Hg and Se caused aberrant DNA methylation at gene loci. Furthermore, DEP at 0.1 ppb caused irreversible heterochromatin changes in ESCs, and DEP-, Hg- and S-421-exposed cells also exhibited impaired formation of the embryoid body (EB), which is an in vitro model for early embryos. We established a system for assessment of epigenetic mutagens. We identified environmental chemicals that could have effects on the human fetus epigenetic status.  相似文献   
33.
This study attempted to establish a culture model to recreate the milk production pathway in bovine mammary epithelial cells (BMECs). BMECs were isolated from Holstein cows (nonlactating, nonpregnant, and parous) and were stored by cryopreservation. To separate the apical and basolateral compartments, BMECs were cultured on a cell culture insert with a collagen gel in the presence of bovine pituitary extract and dexamethasone to induce milk production and tight junction (TJ) formation. The culture model showed the secretion of the major milk components, such as β‐casein, lactose, and triglyceride, and formed less‐permeable TJs in BMECs. Moreover, the TJs were distinctly separated from the apical and basolateral membranes. Glucose transporter‐1, which transports glucose into the cytoplasm through the basolateral membrane, localized in the lateral membrane of BMECs. Toll‐like receptor‐4, which binds to lipopolysaccharide in the alveolar lumen in mastitis, localized in the apical membrane. Beta‐casein was mainly localized near the Golgi apparatus and the apical membrane. Moreover, milk components were almost secreted into the upper chamber of the cell culture insert. These findings indicate that this model has clear cell polarity as well as in vivo and is effective to study of milk production and the blood–milk barrier in lactating BMECs.  相似文献   
34.
This study was conducted to investigate the effect of seven concentrations of Cas9 protein (0, 25, 50, 100, 200, 500, and 1,000 ng/µl) on the development and gene editing of porcine embryos. This included the target editing and off‐target effect of embryos developed from zygotes that were edited via electroporation of the Cas9 protein with guide RNA targeting Myostatin genes. We found that the development to blastocysts of electroporated zygotes was not affected by the concentration of Cas9 protein. Although the editing rate, which was defined as the ratio of edited blastocysts to total examined blastocysts, did not differ with Cas9 protein concentration, the editing efficiency, which was defined as the frequency of indel mutations in each edited blastocyst, was significantly decreased in the edited blastocysts from zygotes electroporated with 25 ng/µl of Cas9 protein compared with that of blastocysts from zygotes electroporated with higher Cas9 protein concentrations. Moreover the frequency of indel events at the two possible off‐target sites was not significantly different with different concentrations of Cas9 protein. These results indicate that the concentration of Cas9 protein affects gene editing efficiency in embryos but not the embryonic development, gene editing rate, and non‐specific cleavage of off‐target sites.  相似文献   
35.
For decades, seedling peaches have been used as the standard rootstock in California almond orchards. Vigorous, deep rooted trees are needed in almond orchards for maximum yields and to withstand the annual tree-shaking at harvest. Currently, researchers are actively evaluating rootstocks for almonds in field trials and in various screening protocols. In this study, seedling rootstocks, obtained from male-sterile advanced generation peach-almond (PEAL) hybrid mother trees, were compared with ‘Nemaguard’ peach seedlings for emergence in the nursery row, trunk caliper at propagation time, and end of season dormant above ground tree weight. Seedling emergence was affected significantly (P ≤ 0.05) by seed source, as was trunk caliper and end of season dormant above ground tree weight. Trunk caliper and dormant above ground tree weight were also affected significantly (P ≤ 0.05) by planting year. Results obtained in this study demonstrate the enhanced first year growth from seedlings of advanced generation PEAL hybrids, as compared to ‘Nemaguard’ seedlings. Seedling emergence in the rootstock bed was not affected significantly by planting year. Advanced generation PEAL hybrid seedlings were ready for June-budding at an earlier date compared to ‘Nemaguard’ seedlings, providing the potential for larger-sized finished nursery stock by the end of the growing season. Due to the male-sterile status of the advanced generation PEAL mother trees, bloom periods of several root-knot nematode resistant rootstock cultivars were examined for their degree of synchronicity with the mother trees. The examined rootstocks and mother trees varied in both chill hour and post-chill heat requirements necessary to effect bloom. ‘Flordaguard’ peach rootstock began bloom in advance of the male-sterile mother trees, whereas the bloom period of ‘Tsukuba No. 4’ occurred well after, suggesting they would not be effective synchronous pollenizers for consistent hybrid seed production. Based on more limited flowering period data, better bloom synchronicity was achieved with a Tsukuba No. 4 X Flordaguard hybrid.  相似文献   
36.
Greenhouse trials were carried out in order to test the efficacy of different seed treatments as alternatives to chemicals against Colletotrichum lindemuthianum cause of anthracnose on bean and Ascochyta spp. cause of Ascochyta blights on pea, respectively. Resistance inducers, commercially formulated microorganisms, non-formulated selected strains of different microorganisms (fungi, bacteria and yeasts) and plant extracts were applied as dry or liquid seed treatments on naturally infested seeds. Seedling emergence and disease incidence and/or severity were recorded. Almost all seed treatments turned out to be ineffective in controlling the Ascochyta infections, which is in line with the literature stating that these pathogens are difficult to control. The only alternative treatments that gave some control of Ascochyta spp. were thyme oil and a strain of Clonostachys rosea. The resistance inducers tested successfully controlled infections of bean by C. lindemuthianum. Among the formulated microorganisms, Bacillus subtilis-based formulations provided the best protection from anthracnose. Some strains of Pseudomonas putida, a disease-suppressive, saprophytic strain of Fusarium oxysporum and the mustard powder-based product Tillecur also proved to be effective against bean anthracnose. However, among the resistance inducers as well as among the other groups, certain agents caused a significant reduction of plant emergence. Different alternative seed treatments can therefore be used for the control of C. lindemuthianum on bean, while on pea only thyme oil and a strain of Clonostachys rosea showed some effectiveness against Ascochyta spp.  相似文献   
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Feeding systems such as grazing affect the fatty acid profile of bovine milk fat. In addition, milk fat is formed as the product of fatty acid metabolism in cow bodies before being secreted into milk. However, how grazing influences milk fatty acid profile through the metabolism has not been completely characterized. When fatty acid concentrations in Holstein milk were compared between grazing and non‐grazing periods, α‐linolenic acid was significantly higher in the grazing period than in the non‐grazing period. This could be explained with an increase in α‐linolenic acid feeding with grazing. α‐linolenic acid had a linear positive correlation with conjugated linoleic acid (9c,11t‐18:2) (CLA) and vaccenic acid (VA) during the grazing period, whereas CLA had higher correlation with linoleic acid rather than with α‐linolenic acid during the non‐grazing period. These data indicate that the high content of dietary α‐linolenic acid affects CLA and VA formation in milk of grazing periods via α‐linolenic acid metabolism into VA.  相似文献   
40.
Recently, we established the GEEP (“gene editing by electroporation of Cas9 protein”) method, in which the CRISPR/Cas9 system, consisting of a Cas9 protein and single guide RNA (sgRNA), is introduced into pig zygotes by electroporation and thus induces highly efficient targeted gene disruption. In this study, we examined the effects of sgRNA on the blastocyst formation of porcine embryos and evaluated their genome‐editing efficiency. To produce an animal model for diabetes, we targeted PDX‐1 (pancreas duodenum homeobox 1), a gene that is crucial for pancreas development during the fetal period and whose monoallelic disruption impairs insulin secretion. First, Cas9 protein with different sgRNAs that targeted distinct sites in the PDX‐1 exon 1 was introduced into in vitro‐fertilized zygotes by the GEEP method. Of the six sgRNAs tested, three sgRNAs (sgRNA1, 2, and 3) successfully modified PDX‐1 gene. The blastocyst formation rate of zygotes edited with sgRNA3 was significantly (< 0.05) lower than that of control zygotes without the electroporation treatment. Our study indicates that the GEEP method can be successfully used to generate PDX‐1 mutant blastocysts, but the development and the efficiency of editing the genome of zygotes may be affected by the sgRNA used for CRISPR/Cas9 system.  相似文献   
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