Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.     

油菜籽来源抗氧化物质研究进展

油菜籽中含有多种具有抗氧化活性的物质，为抵御机体的氧化损伤起到了积极作用。本文综述了油菜籽来源的诸多抗氧化活性成分及抗氧化活性的评价方法。油菜籽主要的抗氧化成分有肽类、多酚类、多糖类、生育酚和其它抗氧化成分，其中的肽类主要有三种产生方式：非消化酶酶解、消化酶酶解和发酵。针对油菜籽来源的活性物质抗氧化活性的评价方法，包括体外测定方法（化学检测）和细胞内抗氧化实验和动物实验。综述还探讨了油菜籽抗氧化活性成分研究中存在的问题和解决方法，旨在为油菜籽活性成分的开发和利用提供参考。     

红曲发酵力测定方法的可行性研究

红曲米是福建红曲黄酒酿制过程中重要的糖化发酵剂。发酵力是衡量酿造用红曲品质的一个重要指标。研究通过酒精测定法和二氧化碳失重法测定酿造用红曲的发酵力,以评价2种发酵力测定方法的可行性。结果表明,二氧化碳失重法更适合于红曲米发酵力的测定。研究还对福建省20个红曲米的发酵力进行了测定,结果表明用二氧化碳生成量能够很好地表征红曲米发酵力的强弱。红曲米发酵力测定方法可行性分析对红曲米相关标准的制定具有重要意义。     

Changes of Morphological Parameter of Wheat Seed Pretreatment by the Biocontrol Agents Bacillus subtilis QM3 during Germination

Bacillus subtilis QM3, a potential biocontrol agent and useful plant-growth-promoting bacteria (PGRB) from yak dung in China with broad-spectrum and strong antifungal bioactivity, was studied to determine if it either stimulated or inhibited the germination of wheat seeds depending on the amount of bacteria. Germination rate (GR) and length of shoot (LS) increased at 10-fold dilution of QM3 fermentation liquid for 12 h (P < 0.05). Germination potentiality (GP) and root cap ration (RC) decreased at 100-fold dilution of QM3 for 0 h. Length of root (LR) and tips of root (TR) at day 7 remained at control levels at 0-fold dilution of fermentation liquid of QM3 for 24 h. The results obtained suggest that some plant diseases, environmental stresses (such as high salt and heavy metal contents), and difficulties in germination can be controlled or relieved by the use of QM3.     

Determination of vc and mating types of Cryphonectria parasitica isolates by multiplex PCR and their genetic diversity in 13 chestnut-growing provinces of Turkey

Vegetative compatibility (vc) and mating types and genetic diversity of Cryphonectria parasitica isolates were determined using 183 isolates obtained from 215 infected chestnut trees growing in 13 provinces of Turkey. Based on the cultural aspects, 143 of these isolates were evaluated as virulent whereas the remaining 40 isolates were hypovirulent. When vc types of 183 isolates were classically differentiated, 135 of them matched to EU-1 (82.3%), 29 of them to EU-12 (17.6%) vc type, whereas 19 of them did not match to the two. When molecular vic markers were used, all the isolates were assigned to two EU vc types; 149 to EU-1 (81.4%) and 34 (18.5%) to EU-12. Of the majority of the isolates, 134 (73.2%) had mating-type MAT-1, while 44 (24%) isolates had MAT-2 and 5 (2.8%) isolates had both mating types. The population analysis based on two DNA marker systems, Inter-Primer Binding Site and Start Codon Targeted Polymorphism, showed no intraspecific genetic variation among the C. parasitica isolates. The prevalence of two dominant vc types revealed by this study shows that biological control with hypovirulent EU-1 and EU-12 isolates will be significant for the country. The results might be helpful to chestnut breeders carrying out resistance breeding studies to manage this disease based on hypovirulence attributed to Cryphonectria hypovirus 1.     

鲤病毒病原的感染性测定

为查明养殖鲤（Cyprinus carpio）突然大批发病死亡的原因，对鲤病样品进行了细胞攻毒、空斑测定、电镜观察，以及鱼体感染等实验。先以患病鲤的组织匀浆液，经过滤后，分别接种到草鱼鳍条细胞（GCF）、鲤上皮瘤细胞（EPC）等14种培养细胞中。利用倒置显微镜观察显示，在1-2d内，该病鱼组织匀浆液可使其中9种细胞出现典型的细胞病变。收集出现病变的细胞液（即病毒悬液），进一步进行病毒滴度检测、空斑测定和鱼体感染实验。结果显示，在GCF细胞上的病毒滴度为10^7.3TCID50／mL；在FHM，TSB和GCO等细胞中可产生直径1～4mm的圆形空斑，空斑的大小与宿主细胞的种类和接种的病毒浓度有关。通过对感染细胞制备的超薄切片和病毒负染样品进行电镜观察，显示这是一类呈典型子弹头样的弹状病毒颗粒。感染了病毒悬液的鲫和鲤先后在第2天和第3天开始出现病症，间隔1～2d后发病的鱼开始死亡，至第14天，两种感染鱼的死亡率均达到83．3％。收集人工感染后濒死的鲫和鲤，分别制备组织匀浆液，回接感染鱼类培养细胞，24h内能使其出现与原发病鲤组织匀浆液所引起的类似的细胞病变。因此证实患病鲤是由病毒病原感染所致。[中国水产科学，2006，13（4）：617—623]     

应用酵母双杂交方法筛选与猪瘟病毒Npro蛋白相互作用的宿主蛋白

为研究猪瘟病毒(CSFV)与宿主细胞间的相互作用,本研究采用酵母双杂交方法以猪瘟病毒Npro蛋白为诱饵,从猪外周血单个核细胞(PBMC) cDNA表达文库中筛选与之相互作用的蛋白,经共转化试验表明,共筛选到12个与Npro蛋白相互作用的宿主蛋白,应用Cytoscape 2.8.2软件绘制了蛋白质间相互作用关系图,根据GO分析结果,这些蛋白分别参与细胞代谢、细胞生长、免疫等过程,经双分子荧光互补(BiFC)试验表明筛得的部分宿主蛋白与CSFV Npro具有相互作用.本研究中筛选得到的蛋白质对CSFV复制的影响有待进一步深入研究.     

传染性脾肾坏死病毒_ConA和L_省略_对体外鳜的头肾淋巴细胞转化的影响

采用MTT颜色反应法研究传染性脾肾坏死病毒（ISKNV）、伴刀豆球蛋白A（concanavallin A,ConA)和脂多糖(lipopolysaccharide,LPS）在离体状态下对健康和ISKNV感染后存活30d或60d的鳜的头肾淋巴细胞转化的影响。结果表明，ConA和LPS能促进健康鳜头肾淋巴细胞的转化，而对感染后存活30d或60d的鳜头肾淋巴细胞没有作用；纯化的ISKNV对健康鳜头肾淋巴细胞没有促进淋巴细胞转化的作用，但对ConA的作用有抑制，对感染后存活的30d或60d的鳜头肾淋巴细胞，纯化的ISKNV有一定的促进转化的作用，感染存活的鳜头肾中有对ISKNV的免疫记忆性T细胞。另外，用建立的ISKNV PCR检测方法对健康和ISKNV感染后存活的鲜组织进行了检测，结果显示，健康鳜为阴性，而ISKNV感染后存活的鱼，头肾、后肾、脾脏和部分鱼的心脏为阳性。ISKNV在鳜体内形成潜伏感染。     

中华鳖同工酶研究

采用垂直淀粉胶电泳对中华鳖脑、眼、心、肌、肝、肾和卵巢七种组织中的六种同工酶系统（ＬＤＨ、ＡＤＨ、ＩＤＨ、ＭＥ、Ｇ６ＰＤＨ、ＥＳＴ）进行研究，构建了中华鳖同工酶酶谱，并利用草鱼ＬＤＨ－Ｂ４抗体对中华鳖ＬＤＨ酶谱进行了免疫化学分析，确定了中华鳖ＬＤＨ同工酶酶谱和亚基组成，揭示了鱼类ＬＤＨ与爬行类ＬＤＨ在进化上的保守性