PEDV N蛋白的原核表达纯化及B细胞抗原表位预测分析

本研究旨在对猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)N基因进行克隆构建、表达与其B细胞抗原表位性质的预测。将PEDV的N基因进行PCR扩增,克隆到原核表达载体pET-28a(+)中,经酶切验证获得阳性克隆,将阳性克隆在表达菌E. coli BL21(DE3)中表达。通过对表达条件及纯化条件的优化,获得高纯度表达蛋白。利用生物信息同源模型化软件Swiss-Pdb Viewer建立PEDV-N蛋白的3D结构,并根据生物信息学软件DNA-Star预测PEDV-N蛋白的二级结构、抗原性、亲水性和表面可及性分析,综合分析预测其B细胞抗原表位。经预测PEDV-N蛋白氨基酸序列中存在13个B细胞优势抗原表位区域。研究结果将进一步为PEDV-N蛋白体外表达产物的应用及研制基因工程疫苗提供理论基础。     

转Cry抗虫基因玉米对家蚕的安全性评价

家蚕可能通过取食漂移到桑叶上的玉米花粉从而受到转基因玉米中表达的Cry蛋白的影响。实验采用室内生物测试的方法评价了转cry1Ab和cry2Ab基因玉米GAB-3花粉对家蚕可能产生的影响,并采用在饲料中掺入蛋白的方式检测了转基因玉米中广泛使用的6种Cry杀虫蛋白对家蚕的毒性。当桑叶上GAB-3花粉密度为50粒·cm^-2时,对家蚕幼虫存活率无显著影响,达500粒·cm^-2时,家蚕14 d存活率为0;以10粒·cm^-2的GAB-3花粉饲喂家蚕幼虫7 d对其体重无显著不利影响,但延长饲喂时间或者当GAB-3花粉密度为50粒·cm^-2时,家蚕的体重显著低于对照组;转基因和非转基因玉米花粉处理14 d或更长时间的家蚕体重均低于无花粉处理组。与非转基因对照处理组相比,低于100粒·cm^-2的转基因玉米GAB-3花粉处理组家蚕的化蛹率、茧重、茧壳重和蛹重无显著差异,转基因玉米和非转基因玉米花粉处理均会延长家蚕幼虫的发育历期。GAB-3花粉对家蚕7 d的LC₅₀为261.18粒·cm^-2,随着处理时间延长,LC₅₀下降。测试的6种Bt杀虫蛋白对家蚕的毒性依次为Cry1C>Cry1Fa>Cry1B>Cry2A>Cry1Ab>Cry1Ac。目前,在中国接近商业化的转基因玉米中多采用的Bt杀虫蛋白为Cry1Ab和Cry2A,二者对家蚕的毒性较低,并且在实际生产中,家蚕接触到的转基因玉米花粉密度较低,因此转Cry抗虫基因玉米对家蚕的风险较低。     

湿度、NaCl浓度对铜仁珍珠豆型花生种子活力及发芽的影响

研究发芽床不同含水量及NaCl浓度对铜仁珍珠豆型花生种子活力及发芽的影响,试验表明,发芽床含水量从20%升高至40%,铜仁珍珠豆型花生种子的发芽率、发芽势、发芽指数呈上升趋势,含水量继续升高至60%,三项指标呈下降趋势,40%是铜仁珍珠豆型花生最适发芽床含水量,花生发芽各项指标达到最高。用NaCl溶液处理种子,浓度从0mol/L升高至0.5mol/L,花生种子的发芽率、发芽势、发芽指数三项指标表现出先急剧后缓慢又急剧下降的趋势。     

太子参须提取物对鸭流感病毒体外试验初探

为尝试性探究太子参须提取物对鸭H9N2亚型流感病毒体外生长的抑制作用,本文基于TCID50试验检测鸭H9N2亚型流感病毒体外感染不同处理组的MDCK细胞.结果 表明,太子参须提取物高低剂量组(75％、25％)和黄芪多糖阳性对照组均对鸭H9N2亚型流感病毒体外生长产生抑制,且太子参须提取物高剂量组(75％)的抑制作用最优...     

水文分异条件下光养生物膜藻类对氮/磷的响应

光养生物膜是河流生态系统中重要的初级生产者,在河流生物地球化学循环中扮演重要角色。然而,河流水文状态及营养盐差异引起的生境异质性对光养生物膜藻类的影响未知。本研究在微型跑道池模拟流水(0.5 m/s)和静水(0 m/s)条件的基础上,通过设置不同浓度氮(1.51, 2.51和5.51 mg/L)、磷(0.1, 0.2和0.4 mg/L)及氮磷比(8, 16和32)条件培养野外采集的原位生物膜及人工基质,探究水文分异和营养变化对河流光养生物膜藻类物种组成及密度的影响。非度量多维排序(NMDS)分析结果表明,差异水文条件下,原位和建群生物膜藻类群落在梯度氮、磷和氮磷比环境中均呈现明显分离(PERMANOVA,P < 0.001),且建群生物膜中各分组藻类群落具有更明显的差异。双因素方差分析结果表明,生物膜中硅藻、蓝藻和绿藻的生长对营养盐与水文变化的响应并不一致,其中磷处理中,磷与流速单一及其交互效应显著影响了大多数藻类的生长及建群(P < 0.001);而氮处理中,氮与流速的交互效应仅对建群生物膜藻类影响显著(P < 0.001)。研究结果也发现,藻类在静水环境更有利于建群生物膜的形成,且静水-高营养盐环境更有利于蓝藻和绿藻的生长。这些结果说明,生物膜建群初期易受到水文扰动的影响,且水文分异和氮、磷影响了光养生物膜藻类的响应模式,研究为河流生态保护提供了新视角。     

Median Lethal Concentration (LC50) of Piscicidal Plants and Their Utilization in Aquaculture

Aqueous and ethanol extracts of Acacia pennata (bark), Catunaregam uliginosa (fruit), Diospyros lenceofolia (fruit), and Sapindus mukorossi (fruit) were evaluated as piscicides on the freshwater fish Danio dangila, Danio rerio, and Puntius shalynius, and the air-breathing Heteropneustes fossilis. Catunaregam uliginosa (fruit) possessed relatively higher piscicidal potency followed by Sapindus mukorossi (fruit), Acacia pennata (bark), and lastly Diospyros lanceofolia (fruit). Catunaregam uliginosa can thus be used for eradicating unwanted wild fishes in a short time span at low concentrations so that the health of aquatic environment is maintained.     

氟离子对西伯利亚鲟胚胎发育的影响

本试验在水温17±1℃的条件下,采用半静态式生物毒性试验方法,探讨了氟离子(F-)对西伯利亚鲟胚胎发育的影响。结果表明,100~600 mg/L氟暴露会导致胚胎孵化延迟,氟暴露组半数孵化时间(MHT)比对照组推迟9~22 h。氟暴露同时导致胚胎死亡率显著增加(P<0.05),出膜仔鱼活力减弱、存活时间缩短、畸形率显著升高(P<0.05),出现卵黄囊畸形、脊椎畸形、眼部充血等畸形症状。试验得出F-致西伯利亚鲟胚胎144 h半致死浓度(LC50)为447.61 mg/L,孵化安全浓度(SC)为4.48 mg/L,导致仔鱼畸形的半数效应浓度(EC50)为536 mg/L。氟对西伯利亚鲟胚胎的安全质量浓度低于我国部分高氟地区地下水、地表水及人为氟污染水域中氟含量,提示天然水体中氟可能会对鱼类胚胎产生负面影响,应引起重视。     

Seminal plasma proteins of Atlantic halibut (Hippoglossus hippoglossus L.)

Protein content and properties in the seminal plasma of Atlantic halibut (Hippoglossus hippoglossus) were assayed using spectrophotometric and electrophoretic methods. The protein concentration ranged from 6.4 ± 3.1 to 19.4 ± 3.4 mg ml⁻¹ and anti-proteolytic activity from 585.2 ± 104.6 to 2912.4 ± 367.4 U l^−l. A high correlation between anti-proteolytic activity and protein concentration (r = 0.95), and between sperm concentration and osmolality was found (r = 0.92). There was a significant decrease in anti-proteolytic activity from the first to the second sampling, but not in protein concentration. Anti-proteolytic activity and protein concentration were significantly affected by variations in individual males. Electrophoresis revealed four anti-proteolytic bands and individual differences in bands of proteolytic activity, which were subsequently characterized as metalloproteases and serine proteases.     

Agronomic Practices Influence Maize Grain Quality

Abstract

Maize grain yields have increased dramatically over the past fifty years, and concurrently end-uses have proliferated requiring special quality characteristics. Plant breeders have developed many specialty types of maize, all of which are influenced by the agronomic practices used to produce the crop. Grain yield increases have resulted in lower protein concentration except when the yield increase resulted from nitrogen fertilizer application. Irrigation improves the biological value of protein, while higher nitrogen application rates alter the amino acid balance thereby reducing the nutritional value. Kernel breakage susceptibility and kernel density increase with higher nitrogen fertilizer application rates, and are reduced by irrigation. Extractable starch and oil concentration are largely influenced by hybrid choice, but small production practice effects have been documented. Essential mineral nutrient levels are often influenced by soil or foliar fertilizer application. The production of aflatoxin can be reduced by irrigation or other strategies to avoid water stress during grain fill. Although genetics usually exerts the largest effect on maize grain quality, agronomic practices are also important.     

晚播条件下施氮量对稻茬小麦氮素吸收及产量的影响

为给安徽省江淮稻麦轮作区域晚播稻茬小麦高产栽培的氮肥合理施用提供依据,选用当地主栽品种扬麦18和皖垦麦076为试验材料,设置5个施氮水平(0、90、180、270和360 kg·hm~(-2)),分析施氮量对晚播小麦氮素积累与分配、糖氮比、氮素同化酶活性及产量的影响。结果表明,增施氮肥能显著提高小麦各器官的氮积累量以及营养器官花前贮存氮素转运量、转运效率和转运氮素对籽粒氮素的贡献率,氮素收获指数随着施氮量的增加而降低。随施氮量的增加,小麦各生育时期不同器官的糖氮比值显著降低。小麦的氮素分配比例在生育前期以叶片最高,成熟期籽粒中氮素分配比例显著高于其余部位,而小麦的可溶性糖分配比例在生育前期以茎鞘最高,成熟期籽粒较高。在0～270 kg·hm~(-2)施氮量范围内,增施氮肥后,两个小麦品种的硝酸还原酶、谷氨酰胺合成酶和谷氨酸合成酶活性均显著提高,穗数、穗粒数和籽粒产量均明显增加。继续增加施氮量至360kg·hm~(-2)时,氮素同化酶活性和产量无显著变化,说明施氮过多对小麦氮素同化和产量无益。土壤氮贡献率、氮肥农学利用效率和氮素偏生产力均随施氮量增加而降低。推荐江淮区域稻茬小麦晚播条件下适宜施氮量为180～270kg·hm~(-2)偏下限,可兼顾高产及氮素高效吸收和利用。