Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

青藏高原地区积雪变化及特征分析

利用我国高原地区1963-2002年的逐月积雪日数资料,以及1972-2011年的格点积雪月频率资料,分析、对比了两种资料在该地区的时空演变特征。结果表明：同时段的(1972-2002)站点资料与格点资料相比,空间分布其高值区与积雪日数增值区均偏西北,偏北程度大致为3~4°,偏西程度大致为2~3°,且中心更为明显。在青藏高原西侧(85°E以西),积雪日数呈整体下降趋势。两种资料的积雪日数EOF分析存在一定的偏差,这与其对应的空间分布差异及气候趋势空间分布存在的偏差相吻合。     

甜菜BvGSTU9基因的生物信息学及在镉胁迫下的表达分析

为了进一步研究甜菜谷胱甘肽转移酶BvGSTU9 (LOC104894060)在重金属胁迫过程中的功能。本研究以‘780016B/12优’为实验材料,对该基因序列特征、结构、功能进行预测分析,并利用qPCR检测该基因在不同浓度镉胁迫下的表达量变化。结果显示甜菜BvGSTU9基因全长925 bp,开放阅读框675 bp,编码了由224个氨基酸组成的不稳定膜外蛋白。BvGSTU9与菠菜、藜麦的氨基酸序列相似性较高,与系统发育进化树分析结果基本相符。二级和三级结构预测表明该基因主要由α-螺旋、β-折叠、延伸链及无规则卷曲组成。qPCR显示BvGSTU9基因在不同浓度的镉胁迫下均受到不同程度的诱导,因此可以推断甜菜BvGSTU9基因无论从结构还是功能上,与镉逆境胁迫存在着一定的应答关系。研究结果也为甜菜耐重金属镉机制研究提供参考依据。     

Effects of Altitude Change on Nutritional Quality and Elymus nutans Regularity in Qinghai-Tibet Plateau

In order to reveal the effects of different altitudes on the nutritional quality of wild forage in Qinghai-Tibet Plateau,this study used Elymus nutans as the tested plants to evaluate the changes of nutrient contents and the correlation between the nutrient contents and altitudes.Four sampling sites were set up at altitudes of 2 562,2 660,2 775 and 2 905 m with the vertical distance among each sampling point was about 100 m.The results showed that the crude protein,ash and phosphorus contents of Elymus nutans decreased significantly with an increase in altitudes.The crude protein contents decreased by 1.87 % with each 100 m increase in altitudes.The variation rule of crude protein contents with altitudes in the simulated curve was "y=–0.0187x+63.244 (R~2=0.9557)".The crude ash contents decreased by 1.77% with each 100 m increase in altitudes.The variation rule of crude ash contents with altitudes in the simulated curve was "y=–0.0177x+56.144 (R~2=0.978)".Neutral detergent fiber (NDF) and acid detergent fiber (ADF) showed an overall increasing trend with the increase of altitudes,but the most obvious increase was at 2 775 m.The contents of crude fat and Ca did not change regularly with altitudes.     

基于BP神经网络的人机联合率定SWMM研究

为解决雨洪管理模型(SWMM)在率定过程中参数复杂、过程繁琐等问题。以西宁市某地块为例建立SWMM模型,利用Morris筛选法进行参数灵敏度分析,并根据灵敏度分析的结果进行人工率定;另外利用BP神经网络对模型进行率定,并结合参数灵敏度对其进行优化。对3种率定方案进行分析,结果表明:水文水力模块参数的相对灵敏度基本一致,其中灵敏度较大的参数为子汇水区面积(Area)、不透水率(Imperv)和不透水区洼地蓄积量(Destore-Imperv),并且不同降雨条件下模型参数的灵敏度存在差异。经过优化后的BP神经网络参数率定方法的模型模拟效果最好,纳什系数最大。结合灵敏度优化BP神经网络的人机联合率定方法一方面能提高BP神经网络率定的准确性,另一方面又能提高传统人工率定的效率。     

人工饲粮对幼年工蜂生长发育及生理代谢的影响

本试验利用液相色谱-质谱联用技术(LC-MS)对幼年工蜂进行代谢组学分析,旨在探究不同人工饲粮对幼年工蜂生长发育及生理代谢的影响,为蜂王邮寄的人工饲粮配制提供一定理论依据。试验选取1日龄的意大利蜜蜂(Apis mellifera ligustica)300只,分为5组,分别饲喂5种人工饲粮,每组30只工蜂。对照组饲粮:蔗糖粉∶蜂蜜=3∶1;试验组A饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶5∶1;试验组B饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶3∶1;试验组C饲粮:蔗糖粉∶花粉∶蜂蜜=6∶5∶1;试验组D饲粮:蔗糖粉∶花粉∶蜂蜜=6∶3∶1。用5种人工饲粮进行室内喂养,记录蜜蜂7 d内的死亡数量;解剖试验组A和对照组第1、3、5、7天的工蜂咽下腺,测定其平均重量、颜色和饱满度;采用LC-MS方法检测试验组A和对照组蜜蜂饲喂7 d后的代谢物差异,对检测数据进行模式识别分析,筛选并鉴别差异代谢物。结果表明:1)对照组和试验组A蜜蜂第1~5天时均没有出现死亡,在第6和7天时试验组A蜜蜂死亡数量均显著低于对照组(P<0.05)。试验组B、C、D的蜜蜂在6 d内全部死亡。2)试验组A第7天时的蜜蜂咽下腺平均重量显著高于对照组(P<0.05),咽下腺颜色为全部乳白色,且咽下腺小体饱满。3)采用主成分分析(PCA)、偏最小二乘法判别方法(PLS-DA)和正交偏最小二乘方-判别分析(OPLS-DA)分析代谢组数据,结果显示试验组A和对照组明显分离,共鉴定出23个差异代谢物,包括氨基酸、脂类、糖类等,其中有10种代谢物上调,13种代谢物下调。差异代谢物通路分析发现,差异极显著的通路有牛磺酸和亚牛磺酸生物合成通路、赖氨酸降解通路、戊糖磷酸盐代谢通路、戊糖和葡萄糖醛酸相互转化通路(P<0.01),差异显著的通路有氨基酸的生物合成通路,色氨酸代谢通路,碳代谢通路,丁酸代谢通路,丙氨酸、天冬氨酸和谷氨酸代谢通路,丙酮酸代谢通路(P<0.05)。由此可见,试验组A的饲粮更适合蜜蜂工蜂的生长发育和生存,在邮寄环境中可代替常用的炼糖饲粮。     

小麦山农4143抗黑胚病遗传分析及抗性遗传位点检测

 黑胚病是小麦生产的重要籽粒病害,麦根腐平脐蠕孢(Bipolaris sorokiniana)是黑胚病的主要致病菌。为分析小麦抗黑胚病遗传规律并检测抗性位点,本研究以抗黑胚病小麦品系山农4143与感病品系宛原白1号的F₇代重组自交系(RIL)群体为材料,于2018~2019年在3个试验点种植,采用“孢子液喷洒、套袋保湿”的方法进行接菌鉴定,用“主基因＋多基因混合遗传模型”进行遗传分析,并挑选极端抗、感自交系构建混池、结合小麦660K SNP芯片进行混合分组分析(BSA)。研究结果显示小麦品系山农4143对黑胚病抗性符合“4对主基因加性-上位性遗传模型”,主基因遗传力为0.88~0.95;通过BSA检测到黑胚病抗性位点36个,分别位于1A、2B(6)、2D(2)、3A、3B(2)、3D、4A(6)、4D、5A(4)、5B(6)、5D(3)、7A、7B和7D共14条染色体上,A、B、D染色组上分别为13、 15和8个,36个位点中有18个为抗黑胚病新鉴定位点。本研究结果为小麦抗黑胚病位点的精细定位和抗性育种中分子标记的开发奠定了基础。     

箭筈豌豆、玉米产量对间作和施氮水平的响应

研究不同施氮水平下箭筈豌豆、玉米产量对单作和间作的响应,对于优化栽培措施、提高作物产量具有重要意义。本试验于2017年在甘肃河西绿洲灌溉区进行,重点研究是不同施氮水平下箭筈豌豆、玉米在单作及间作模式中干物质累积、产量构成、产量表现。结果表明,间作箭筈豌豆干物质累积量与单作相比在不施氮(N0)、减量施氮(N1)和常规施氮(N2)处理下分别提高44.0%、36.7%和37.2%;施氮水平间,间作模式下N1比N2和N0提高9.1%和25.5%,单作模式下提高9.4%和32.1%。间作玉米干物质累积量与单作相比在N0、N1和N2处理下分别提高23.3%、22.5%和23.0%;施氮水平间,间作模式下N1比N2和N0提高9.3%和19.8%,单作模式下提高9.9%和20.7%。间作模式下箭筈豌豆豆荚数和玉米穗数较相应单作分别增加7.6%和14.5%,两者的产量较单作分别提高34.9%和27.0%。由通径分析可知,箭筈豌豆产量决定次序是豆荚数>粒重>单荚粒数,玉米产量决定次序是穗数>粒重>穗粒数。间作结合施氮量240 kg/hm²能获得较高干物质积累量和产量,是河西灌溉区玉米间作箭筈豌豆适宜的施氮水平。     

两种可食用玫瑰生物碱代谢物差异分析

以云南滇红玫瑰(ZY)与墨红玫瑰(DH)两种云南主要的食用花卉品种为研究对象,采用超高效液相色谱-串联质谱(UPLC-MS/MS)方法比较两种玫瑰花次生代谢产物的差异性。结果表明,两种花瓣中共检测到82种代谢产物,包括55种氨基酸及其衍生物(67.07%)、16种生物碱(19.51%)、7种酚胺(8.54%)、4种吲哚类生物碱(4.88%),其中滇红玫瑰中谷胱甘肽含量是墨红玫瑰的26.35倍,具有较强的抗氧化能力;而L-天冬酰胺和S-(5'-腺苷)-L-高半胱氨酸却是墨红玫瑰中特有的物质,具有显著的降血压作用。与墨红玫瑰相比,滇红玫瑰15种差异代谢物显著下调,表明墨红玫瑰较滇红玫瑰有较高的营养保健功能,而滇红玫瑰高表达成分使其具有更强的抗氧化能力。