家兔黏膜免疫器官肥大细胞在巴氏杆菌感染中的特征——组织化学与电镜观察

对5例巴氏杆菌感染家兔的空肠与圆小囊肥大细胞（mast cell，MC）进行了组织化学与电镜的观察，发现在细菌感染导致的病变组织边缘、病变组织内血管附近结缔组织中及变性细胞周围组织的上皮下与黏膜下层都出现了大量甲苯胺蓝（Toluidine Blue，TB）阳性肥大细胞。电镜下，MC有的直接与变性细胞密切接触，有的则和血管内皮紧密相触，在此部位的血管内可见有淋巴细胞贴附填充。MC胞浆内充盈大量的特征性颗粒，有时这些颗粒向细胞表面突出形成边缘空隙，即形成所谓的脱颗粒管道，将颗粒内容物逐渐排除细胞外，遗留空腔。     

Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.     

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.     

Amino Acid-Coupled Bromophenols and a Sulfated Dimethylsulfonium Lanosol from the Red Alga Vertebrata lanosa

Vertebrata lanosa is a red alga that can commonly be found along the shores of Europe and North America. Its composition of bromophenols has been studied intensely. The aim of the current study was therefore to further investigate the phytochemistry of this alga, focusing more on the polar components. In total, 23 substances were isolated, including lanosol-4,7-disulfate (4) and the new compounds 3,5-dibromotyrosine (12), 3-bromo-5-sulfodihydroxyphenylalanine (13), 3-bromo-6-lanosyl dihydroxyphenylalanine (14), 3-(6′-lanosyl lanosyl) tyrosine (15) and 5-sulfovertebratol (16). In addition, 4-sulfo-7-dimethylsulfonium lanosol (7) was identified. While, in general, the dimethylsulfonium moiety is widespread in algae, its appearance in bromophenol is unique. Moreover, the major glycerogalactolipids, including the new ((5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic acid 3′-[(6′’-O-α-galactopyranosyl-β-D-galactopyranosyl)]-1-glycerol ester (23), and mycosporine-like amino acids, porphyra-334 (17), aplysiapalythine A (18) and palythine (19), were identified.     

北美栅锈菌和松杨栅锈菌可视化检测体系建立

目的 为实现北美栅锈菌和松杨栅锈菌快速有效的鉴别及鉴定。 方法 本研究根据两种锈菌的28SrDNA基因序列设计若干组LAMP引物。经筛选得到的引物以北美栅锈菌、松杨栅锈菌、图拉斯叉钩丝壳菌、芍药白粉菌、梨胶锈菌、羊肚菌、金针菇的基因组DNA为模板进行LAMP反应并完成特异性检测;首先建立初始LAMP反应体系,再进一步优化LAMP反应体系的组分和反应条件,加入羟基萘酚蓝实现可视化检测;最后确定检测体系灵敏度。 结果 表明,筛选出的引物具有特异性;25 μL北美栅锈菌LAMP检测体系最佳Mg2＋浓度为6 mmol·L−1,最佳内外引物比例为8：1,最佳dNTPs浓度为1.2 mmol·L−1;同样地,25 μL松杨栅锈菌LAMP检测体系最佳Mg2＋浓度为4 mmol·L−1,最佳内外引物比例为6：1,最佳dNTPs浓度为1 mmol·L−1。加入160 μM羟基萘酚蓝（HNB）可清晰地指示反应结果,两种检测体系在61 ℃条件下,分别反应30 min和40 min可实现目视判断结果,且灵敏度分别可达34 fg·μL−1和60 fg·μL−1。 结论 通过建立两种锈菌的可视化LAMP-HNB检测体系可实现对北美栅锈菌和松杨栅锈菌进行区分及鉴定,为快速鉴定和区分重要杨树锈病提供了技术支撑和实践参考。     

补充不同光质对温室黄瓜生长发育、光合和前期产量的影响

 【目的】研究温室黄瓜对不同光质的反应及其反应机理。【方法】对处于弱光条件下的温室黄瓜进行补充红光（650 nm）和蓝光（450 nm）处理,以补充白光为对照,补光时间10 h（7：30－17：30）,测定不同光质对温室黄瓜生长发育、叶绿素含量、叶片光合特性和前期产量的影响。【结果】补充红光处理提高了温室黄瓜冠层日最高温度、日最低温度、株高、叶面积、叶绿素含量、叶片光合速率、干物质产量,促进同化产物向营养器官分配。蓝光处理提高了温室黄瓜叶片的比叶面积、气孔导度,促进同化产物向果实分配,提高了商品果数量和商品果总产量。【结论】补充红光和蓝光由于提高了温室黄瓜叶片的光合速率、叶面积和比叶面积,从而增加了温室黄瓜的干物质产量,补充蓝光促进了同化产物向果实分配,从而提高了产量。     

一株腐败希瓦氏菌的筛选鉴定及其染料脱色研究

[目的]研究一株腐败希瓦氏菌对染料的脱色效果。[方法]从印染废水中筛选到一株对偶氮染料活性艳红X-3B和蒽醌染料活性艳蓝KN-R均具有高效降解作用的菌株,对其进行了菌种鉴定,并分析了该菌在不同培养条件下对两种染料的脱色效果。[结果]生理生化鉴定和16S r DNA序列鉴定结果表明,该菌为希瓦氏菌属腐败希瓦氏菌,命名为S.putrefaciens 4H。该菌在pH=8、37℃下静置培养时,对两种染料的脱色效果最好。该菌6 h内可将50 mg/L活性艳红X-3B完全降解脱色,在含有活性艳红X-3B的LB平板上可形成水解圈,推测其能产生水解活性艳红X-3B的胞外酶;该菌对50 mg/L活性艳蓝KN-R的脱色率48 h时可达81.84%,且脱色时染料先被细胞吸附沉淀后再随着细胞的生长代谢逐渐降解,其脱色机制还有待研究。[结论]该研究为印染废水的处理提供了理论依据。     

高致病性猪蓝耳病防控

随着人们生活方式的变化,养猪户也变得越来越多,不仅能为其带来更多的经济收入,也可提高其生活质量和水平。调查数据显示,很多养殖户在养猪中通常会因很多因素致使猪群感染疾病,倘若不能及时处理,势必会对自身带来较为严重的经济损失,甚至会加大疫病的扩散。该文主要对高致病性猪蓝耳病防控新理念展开分析,并提出一些可行的对策。     

钼蓝比色法测定笃斯越橘成熟果实中还原型抗坏血酸含量及体系优化

以笃斯越橘(Vaccinium uliginosum)成熟果实为试材,以钼蓝比色法测定笃斯越橘果实还原型抗坏血酸(Reduced ascorbic acid,As A)含量进行条件优化。结果表明,3%偏磷酸-乙酸用量为100μL、5%硫酸和5%钼酸铵用量均为200μL、以去离子水补充至1 500μL,30℃干热恒温浴显色40 min,取出室温下放置1 h后,在700 nm下测定,所得数据稳定,准确性适合用于笃斯越橘果实As A含量的测定;测得含量为(40.20±6.23)mg/100 g FW。