基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

羊IL-1Ra基因全长cDNA克隆及生物信息学分析

试验旨在对羊白介素1受体颉颃因子（interleukin-1 receptor antagonist，IL-1Ra）基因进行全长cDNA克隆及生物学分析。根据布鲁氏菌感染羊白细胞层SSH cDNA文库中的IL-1Ra基因序列信息及已知核苷酸序列（GenBank登录号：KC425613.1）设计引物，利用RT-PCR结合RACE方法扩增并克隆IL-1Ra基因，对其进行序列及相关分子特性分析。结果表明，羊IL-1Ra基因全长为1 228 bp，可编码174个氨基酸，含有1个完整的IL-1保守结构域和1个IL-1Ra结构域（PHA02651结构域）。IL-1Ra分子质量为19 765.8 u，等电点（pI）为5.72，其分子式为C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁，且含有信号肽。二级结构分析显示，IL-1Ra分子存在较多的β折叠及受体结合位点，与三级结构预测结果一致，且与人/鼠IL-1Ra分子的空间结构相似度极高，达到90%以上。羊IL-1Ra有IL-1特有的三叶草结构，其酶结合结构域位于TYR⁴⁷至GLU⁶⁶之间。将羊IL-1Ra与牛、虎鲸、宽吻海豚、猪、家犬、人、鼠等14个物种进行蛋白质同源性比对，发现在各物种间存在5个高度同源的半胱氨酸位点。系统进化树分析发现，羊IL-1Ra基因全长cDNA所编码的氨基酸序列同山羊、牛单独形成一个分支。本研究结果为今后深入研究IL-1Ra基因的生物学功能奠定了基础。     

竹柳等4个柳树无性系造纸材性测定分析

对3年生竹柳、苏柳172、苏柳795、苏柳799进行造纸材性测定,结果表明:4个柳树无性系的木材基本密度在0.324~0.368 g/cm3之间,纤维长度在0.866 9~0.928 8 mm之间,纤维宽度在21.349~22.683μm之间。4个柳树无性系木材纤维的长宽比、壁腔比及腔径比等指标均符合造纸原料要求,而且纤维长度、宽度均分布比较均匀。     

响应曲面法优化草莓汁花色苷促溶技术的研究

为了加速溶出草莓汁中的花青毒,以哈尼草莓为试验材料,原花青素含量为检测指标,依据中心组合响应面设计,研究果胶酶处理对草莓中色素物质溶出的影响。结果表明:果胶酶处理的最佳工艺条件为果胶酶添加浓度为(质量比)1.70‰,45℃处理2.87h;方差分析结果表明模型高度显著(P=0.0002),失拟性不显著(P0.1)。该模型可进行草莓汁加工中花青素溶出的分析及预测。通过该技术处理,可以得到花青素含量高、颜色较好的草莓汁。     

Analysis of the independent-and interactive-photo-thermal effects on soybean flowering

Soybean(Glycine max(L.) Merr.) is a typical short-day and warm season plant, and the interval between emergence and flowering has long been known to be regulated by environmental factors, primarily photoperiod and temperature. While the effects of photoperiod and temperature on soybean flowering have been extensively studied, a dissection of the component photo-thermal effects has not been documented for Chinese germplasm. Our objective of the current study was to evaluate the independent- and interactive-photo-thermal responses of 71 cultivars from 6 ecotypes spanning the soybean production regions in China. These cultivars were subjected in pot experiments to different temperature regimes by planting in spring(low temperature(LT)) and summer(high temperature(HT)), and integrating with short day(SD, 12 h), natural day(ND, variable day-length), and long day(LD, 16 h) treatments over two years. The duration of the vegetative phase from emergence to first bloom(R1) was recorded, and the photo-thermal response was calculated. The outcome of this characterization led to the following conclusions:(1) There were significant differences in photo-thermal response among the different ecotypes. High-latitude ecotypes were less sensitive to the independent- and interactive-photo-thermal effects than low-latitude ecotypes; and(2) there was an interaction between photoperiod and temperature, with the effect of photoperiod on thermal sensitivity being greater under the LD than the SD condition, and with the effect of temperature on photoperiodic sensitivity being greater under the LT than the HT condition. The strengths and limitations of this study are discussed in terms of implications for current knowledge and future research directions. The study provides better understanding of photo-thermal effects on flowering in soybean genotypes from different ecotypes throughout China and of the implications for their adaptation more broadly.     

牡丹5个管家基因的克隆及其在系统进化分析中的应用

利用RT-PCR技术从牡丹组7个野生种和1个栽培种中分别克隆得到泛素延伸蛋白基因(ubiquitin,UBI)、素环蛋白基因(cyclophilin,CYP)、肌动蛋白基因(Actin)、葡萄糖六磷酸脱氢酶基因(glucose-6-phosphate 1-dehydrogenase,G6PD)和β–微管蛋白基因(beta-tubulin,TUB)等5个管家基因的编码序列(coding sequence,CDS)。序列比对分析发现,5个管家基因序列均相对保守,其在牡丹7个野生种和1个栽培种中的保守性在99.04%~99.69%。利用5个管家基因分别构建了其在牡丹组8份材料中的系统进化树。5个系统进化树均将8份材料分成两组,即革质花盘亚组和肉质花盘亚组;由于5个管家基因核苷酸序列各自的保守性不同,在区分亚组成员间略有差异。综合分析5个管家基因构建的系统进化树可知,杨山牡丹(Paeonia ostii)和栽培牡丹(P.suffruticosa‘Luoyanghong’)常聚为一支,二者亲缘关系较近,因而推测杨山牡丹可能参与了栽培牡丹的形成;紫斑牡丹(P.rockii)和四川牡丹(P.decomposita)一直聚为一支,推测二者亲缘关系较近;黄牡丹(P.lutea)和紫牡丹(P.delavayi)一直聚为一支,推测这二者亲缘关系较近。狭叶牡丹(P.potaninii)与黄牡丹和紫牡丹分化较早,结合前人研究结果建议将狭叶牡丹作为一个独立的种。     

Ultrastructural impairment of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice

AIM: To investigate the ultrastructural changes of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice. METHODS: BALB/c mice were randomly divided into diabetic group and control group. The expression of insulin and platelet-endothelial cell adhesion molecule-1 (CD31) in islet microvessels was detected by immunohistochemical staining. The ultrastructural changes of islet β cells and islet microvessels were observed under transmission electron microscope. RESULTS: Compared with control group, the number of islet β cells, ratio of β cells/α cells, average number of secretory granules in β cells and insulin expression area per islet in diabetic group were significantly decreased (P<0.01). Besides, diabetic group had fewer microvessels with lower expression of CD31 (P<0.01). Mitochondria in islet microvascular endothelial cells and pericytes in diabetic group were swelling. The basement membrane of islet microvessels became thicker in diabetic group (P<0.01). CONCLUSION: Islet microvascular endothelial cells were impaired in type 1 diabetic mice.     

Establishment and Optimization of INH Epitope Peptide Vaccine for Detection of Antibody

In this study, an indirect ELISA method was established to detect inhibin hormone (INH) epitope peptide vaccine antibody, it would provide oretical reference for the determination of Fine-wool sheep after active immune body INH epitope peptide vaccine antibody. On the basis of the predecessors, using indirect ELISA method to determinate serum INH epitope peptide antibody levels of sheep, and through control different experimental conditions to look for the best experimental conditions. Through explorating the experimental conditions, finally, the testing experiment conditions were determined, which was blocked solution with skimmed milk powder, INH and GnIH synthetic peptides dilution degrees for 20 000 times, the optimum reaction time was 60 min, the best color action time was 15 min. In this experiment, a kind of method to detection antibody in the body after INH active immune sheep was built, it would provide a reference for future research.     

Physiological characteristics of high yield under cluster planting: photosynthesis and canopy microclimate of cotton

Cotton produces more biomass and economic yield when cluster planting pattern (three plants per hole) than in a traditional planting pattern (one plant per hole), even at similar plant densities, indicating that individual plant growth is promoted by cluster planting. The causal factors for this improved growth induced by cluster planting pattern, the light interception, canopy microclimate and photosynthetic rate of cotton were investigated in an arid region of China. The results indicated that the leaf area index and light interception were higher in cluster planting, and significantly different from those in traditional planting during the middle and late growth stages. Cotton canopy humidity at different growth stages was increased but canopy temperatures were reduced by cluster planting. In the later growth stage of cluster planting, the leaf chlorophyll content was higher and the leaf net photosynthetic rate and canopy photosynthetic rate were significantly increased in comparing with traditional planting pattern. We concluded that differences in canopy light interception and photosynthetic rate were the primary factors responsible for increased biomass production and economic yield in cluster planting compared with the traditional planting of cotton.