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1.
A method of authenticating anchovy (Engraulis encrasicholus L.) and gilt sardine (Sardinella aurita) semipreserves (salt-cured and fillets in oil) has been developed by polymerase chain reaction (PCR) followed by sequence and restriction site analysis. The amplification of a fragment of the cytochrome b gene by universal primers produced a 376 base pairs (bp) fragment in all samples analyzed. Digestion of PCR products with XhoI, TaqI, AluI, and HinfI endonucleases yielded species-specific profiles distinguishing anchovy from gilt sardine. Therefore, the restriction length fragment polymorphism (RLFP) technique can be used to determine the species identity of anchovy and gilt sardine in semipreserves.  相似文献   
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Although an antioxidant mechanism has been involved in the beneficial effects of ferulic acid in human diseases, there are few reports on the antioxidant properties of this compound in isolated membranes and intact cells. Here, we evaluated the ability of ferulic acid in inhibiting lipid peroxidation in rat liver microsomal membranes and reactive oxygen species production in NIH-3T3 fibroblasts, induced by both tert-BOOH and AAPH. We also compared its antioxidant efficiency with that of other antioxidants, such as alpha-tocopherol, beta-carotene, and ascorbic acid, added alone or in combination. Ferulic acid acted as a potent antioxidant in our models, being more effective in protecting from tert-BOOH than from AAPH. Moreover, the compound was the most effective among the antioxidants tested. Synergistic interactions were observed when the compound was used in combination with the other antioxidants, suggesting that they can cooperate in preserving physiological integrity of cells exposed to free radicals.  相似文献   
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The US3 protein is a unique protein kinase only present in the Alphaherpesvirinae subfamily of the herpesviruses. Studies performed with several alphaherpesviruses demonstrated that the US3 protein is involved in cytoskeleton modifications during viral infection and displays anti-apoptotic activity. However, the US3 protein of BoHV-5 has not been studied up to now. As reported for other alphaherpesviruses, our results showed that BoHV-5 US3 confers resistance against apoptosis and induces cytoskeletal reorganization leading to cell rounding, actin stress fiber breakdown and cell projections that interconnect cells. The expression of a kinase-dead version of BoHV-5 US3 showed that the anti-apoptotic activity and the induction of cell projections are kinase-dependent whereas kinase activity is not absolutely required for actin stress fiber breakdown. Besides, the kinase-dead version of US3, but not the wild type protein, was found excluded from the nucleus. These results constitute the first report on the BoHV-5 US3 functions, and highlight that there are functional differences and similarities among US3 proteins of different alphaherpesviruses.  相似文献   
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Olive fruits are seriously deteriorated by pre and postharvest damage due to the attack of insects, such as Bactrocera olaea, which strongly alters the quality of olives. Defence response in olive fruits injured both by pathogens and by mechanical damages has been associated with the enzyme β-glucosidase, which specifically hydrolyses oleuropein, producing highly reactive aldehyde molecules. In situ detection of ~-glucosidase activity in olive fruit tissues following injury, which simulates Bactrocera oleae punctures, is reported. The assay was performed in two cultivars showing different degrees of susceptibilities to fly infestation. In both cultivars, the histochemical assay for β-glucosidase showed that within 20 min after the injury, a strong ~-glucosidase activity could be observed in the damaged tissues. Thereafter a progressive enzyme inactivation occurred starting from tissues around the boundary of the injury with decrease of the enzyme activity and stopped after 3 h. Whereas the mass of active cells reached a distance of (300±50) μm from the edge of the injury. Biochemical analyses showed that in extracts of the injured fruit, β-glucosidase activity rapidly increased within 20 min from injury, thereafter decreasing and reaching values comparable with those in intact fruits. Following puncture, the oleuropein contents did not change significantly in the high susceptibility cultivar, whereas it rapidly decreased in the cultivar showing low susceptibility. The results strongly suggest that olive fruits susceptible towards fly infestation could be related to the ability of the oleuropein-degrading-β-glucosidase to produce the highly reactive molecules in the damaged tissues. As a consequence of puncture, high level of peroxidase activity was detected. This feature also suggested that this enzyme could play a key role in the defence response against insect injuries.  相似文献   
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Olive fruits are seriously deteriorated by pre and postharvest damage due to the attack of insects, such as Bactrocera olaea, which strongly alters the quality of olives. Defence response in olive fruits injured both by pathogens and by mechanical damages has been associated with the enzyme β-glucosidase, which specifically hydrolyses oleuropein, producing highly reactive aldehyde molecules. In situ detection of β-glucosidase activity in olive fruit tissues following injury, which simulates Bactrocera oleae punctures, is reported. The assay was performed in two cultivars showing different degrees of susceptibilities to fly infestation. In both cultivars, the histochemical assay for β-glucosidase showed that within 20 min after the injury, a strong β-glucosidase activity could be observed in the damaged tissues. Thereafter a progressive enzyme inactivation occurred starting from tissues around the boundary of the injury with decrease of the enzyme activity and stopped after 3h. Whereas the mass of active celsreached a distance of (300±50)μm from the edge of the injury. Biochemical analyses showed that in extracts of the injured fruit, β-glucosidase activity rapidly increased within 20 min from injury, thereafter decreasing and reaching values comparable with those in intact fruits. Following puncture, the oleuropein contents did not change significantly in the high susceptibility cultivar, whereas it rapidly decreased in the cultivar showing low susceptibility. The results strongly suggest that olive fruits susceptible towards fly infestation could be related to the ability of the oleuropein-degrading-β-glucosidase to produce the highly reactive molecules in the damaged tissues. As a consequence of puncture, high level of peroxidase activity was detected. This feature also suggested that this enzyme could play a key role in the defence response against insect injuries.  相似文献   
9.
Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.  相似文献   
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本试验研究了从"Redhaven"桃果肉中分离得到的两种拮抗酵母菌株L1和L8对三种核果类褐腐病原菌(Monilinia laxa,Monilinia fructicola及Monilinia fructigena)的离体和活体拮抗效果。这两种拮抗菌株是经初步验证能够减轻桃和油桃的褐腐病症状后筛选出的,并最终通过分子生物学及形态学手段鉴定为Aureobasidium pullulans(出芽短梗霉菌)。通过活体接种三种褐腐病菌的试验研究发现,两种拮抗菌的高压灭菌悬液及无菌过滤液对果实褐腐病的发生均无抑制作用,而含有L1和L8菌株的菌悬液可完全抑制M.laxa和M.fructicola引起的褐腐病的发生,并使M.fructigena引起的褐腐病发病率分别降低70%和90%。在另一项试验中,将油桃用两种拮抗菌液处理后接种三种褐腐病菌,然后将其在0℃下贮藏21 d,再在20℃下放置7 d。结果表明,低温抑制了褐腐病的发生,处理和对照果实均无褐腐病的症状。但在20℃下放置7 d后,对照果实的褐腐病发病率均超过了45%,而LI和L8处理分别可以将接种M.fructigena病原菌果实褐腐病的发病率降低89.8%和91.2%,说明两种拮抗菌的处理可完全抑制接种M.laxa和M.fructicola引起的果实褐腐病的发生。两种拮抗酵母菌株的最佳抑菌浓度为108CFU.mL-1,L1在107CFU.mL-1浓度时也具有较好的抑菌效果。L8菌株在107CFU.mL-1浓度时对由M.fructigena和M.fructicola引起的果实褐腐病无抑制活性,处理与对照果实无差异。而对于接种了M.laxa的油桃果实,与对照相比,107CFU.mL-1浓度的L8菌株却可显著降低褐腐病的发病率。尽管在0℃和20℃下贮藏的油桃果实伤口处的L1和L8菌的数量增长较慢,但已经足够控制果实褐腐病的发生。综上所述,本项研究确定了可作为开发采后生物杀菌剂活性成分的两种A.pullulans拮抗菌株,并用于抑制引起核果类作物采后严重经济损失的三种Monilinia真菌。  相似文献   
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