透明颤菌血红蛋白基因(vgb)在大肠杆菌中的高效表达

利用PCR技术将透明颠菌血红蛋白基因（vgb）克隆到融合表达载体pET28a，在大肠杆菌BL21（DE3）表达。重组蛋白在30℃诱导获得可溶表达。利用Ni^2＋亲和层析对重组蛋白进行了纯化，得到重组的透明颠菌血红蛋白，蛋白呈红色。实验现象显示重组蛋白与血红索相结合。紫外光区和可见光区波长扫描分析显示：蛋白粗提物和纯化后的血红蛋白在230nm和413nm都有强吸收峰，初步表明，尽管重组蛋白比天然蛋白多36个氨基酸，重组蛋白仍然具有生理活性。诱导后4h和6h的培养液氨基乙酰丙酸含量分别达到17mg／L和21mg／L。说明重组蛋白的表达明显促进了体内heme合成途径。     

复方中草药"毒菌杀"对禽大肠杆菌及鸡白痢沙门氏菌的体外抑菌试验

观察了复方中草药“毒菌杀”的安全性及其对禽大肠杆菌及鸡白痢沙门氏菌的体外抑菌情况。结果发现“毒菌杀”安全性高，组成“毒菌杀”的各单味中药及其合剂对大肠杆菌的最低抑菌浓度MIC(g/mL)分别为板蓝根0．05、穿心莲0．05、黄芪0．025、黄柏0．05、柴胡0．05、生地0．05、甘草0．05、当归0．025，“毒菌杀”方剂为0．05；对沙门氏菌的MIC分别为板蓝根0．05、穿心莲0．025、黄芪0．025、黄柏0．05、柴胡0．025、生地0．05、甘草0．025、当归0．05，“毒菌杀”方剂为0．025；而牛胆汁对这两种致病菌的最低抑菌浓度分别为2％和1％。说明“毒菌杀”方剂对大肠杆菌、沙门氏菌具有明显的抑制作用。     

鸡蛋消毒方法的对比研究

采用5种消毒方法对鸡蛋蛋壳表面细菌进行杀菌效果试验,结果表明,各处理组之间抑菌效果差异显著(P＜0.05),杀菌效果优劣依次为开水浸烫＞甲醛熏蒸＞过氧乙酸熏蒸＞高锰酸钾浸泡＞过氧乙酸浸泡,开水浸烫操作技术难度较大,容易引起蛋壳破裂,适宜实际生产的消毒方法为过氧乙酸熏蒸和甲醛熏蒸.将消毒处理后的鸡蛋在温度25℃,相对湿度60%～80%贮存20天,开水浸烫处理组鸡蛋仍保持较鲜的品质,其余各组鸡蛋品质较差.结果表明,常规消毒方法(除开水浸烫法)只能抑制蛋壳表面微生物的生长繁殖,不能延缓鸡蛋品质下降达到保鲜鸡蛋的作用.     

耕作方式与长期定位施肥对雨养农田冬小麦产量的调控效应

为了明确西北黄土高原半湿润偏旱区耕作方式与长期定位施肥对冬小麦产量的调控效应,以设在半湿润偏旱区连续12年的耕作与肥料长期定位试验为平台,采用裂区设计,以传统耕作和免耕耕作为主处理,不施肥(CK)、单施无机氮肥(N)、单施无机磷肥(P)、单施有机肥(M)、无机氮磷肥配施(NP)、有机无机肥配施(NMP)为副处理,栽培制度为1年春玉米-3年冬小麦轮作,研究耕作及施肥措施对冬小麦产量的影响及其在生产年型间的变化关系。结果表明,不同年型及耕作方式均以有机无机配施冬小麦产量最高,有机肥单施高于化肥单施,磷肥单施高于氮肥单施。耕作方式间表现为传统耕作高于免耕耕作,年型间表现为丰水年>平水年>干旱年,耕作和施肥方式的增产效果以干旱年最好,平水年和丰水年差异不显著。有机无机配施与传统耕作结合优化了冬小麦冠层温度、叶绿素相对含量等生理指标,提高了光合速率、气孔导度及蒸腾速率,因而改善了有效穗、穗粒数和千粒重等产量性状而使冬小麦增产。在550 mm左右降水量的陇东旱塬雨养农业区,无论何种耕作方式及生产年型,长期采用有机无机或无机氮磷肥配施均表现出持续提高冬小麦产量的良好作用。因此,有机无机配施结合传统耕作是提高陇东半湿润偏旱区冬小麦产量的最佳耕作栽培模式。     

四倍体鸭茅产量及其构成因素的QTL定位

产量相关性状是复杂的数量性状,对鸭茅的单株产量及构成因素进行QTL分析,可提高育种中对产量性状优良基因选择的效率,同时为鸭茅遗传改良、基因克隆及分子标记辅助育种提供理论依据。以四倍体鸭茅“楷模”和“01436”为亲本杂交而成的作图群体为试验材料,于2014年对洪雅、宝兴两个不同生境下鸭茅株高、旗叶长、倒二叶长、旗叶宽、倒二叶宽、茎粗、花序长、分蘖数、单株干重等9个产量相关性状进行了表型鉴定及相关性分析。此外,在已构建的高密度鸭茅分子遗传图谱的基础上,采用MapQTL 5.0进一步对这些性状QTL定位分析。结果表明,绝大多数性状在亲本间呈显著差异且都整体表现出连续变异,符合数量性状遗传的特征;相关性分析表明,大多数产量相关性状均与干重极显著正相关,与单株干重相关性最好的依次为分蘖、株高;QTL分析发现,控制该9个农艺性状的QTL共60个,洪雅38个,宝兴22个,这些QTL分别定位于分子连锁图谱的1、2、3、4、5共5个连锁群上,单个QTL的贡献率为5.7%~24.7%,单个性状QTL个数为2~15个。其中,控制株高、花序长的QTL各12个,控制倒二叶长、茎粗的QTL各4个,控制旗叶宽、倒二叶宽的QTL各2个,控制单株干重的QTL有6个,影响分蘖的QTL为3个,与旗叶长相关的QTL最多15个。     

高寒区轮作模式对当归田土壤特性及药材产量的影响

连作重茬造成当归产量逐年下降。采用药用植物秦艽(QJ)、羌活(QH)轮作当归(DG),以当归连作为对照,通过测定土壤和药材产量指标,利用主成分分析确定各指标权重,并结合隶属综合因子分析和聚类分析,旨在揭示不同药用作物轮作当归对土壤环境与药材产量的影响,为筛选适宜当归栽培的轮作模式提供科学依据。结果表明,不同作物轮作对当归田土壤特性和药材产量均具有显著影响,秦艽轮作当归可有效改善土壤特性,使当归成药期土壤磷和钾含量处于较低水平,而有机质维持在较高水平,对土壤水分的季节调控作用更为显著,与当归连作相比较,当归成药率提高94.10%,药材单根鲜重和药材鲜产量分别提高53.46%和125.56%,而羌活轮作当归土壤pH最高,有机质含量最低,当归连作土壤有机质含量虽高,但土壤富集磷和钾,对土壤水分季节性调控作用最小,当归成药率和药材产量均低。不同轮作模式当归成药率和药材产量依次为秦艽-当归>当归-当归>羌活-当归,综合指数大小依次为秦艽-当归(0.8132)>羌活-当归(0.3315)>当归连作(0.2051)。聚类分析将3种轮作模式分为三大类,重复小区亚类间距离均小,说明不同作物轮作对土壤的调控作用不同,高寒产区秦艽轮作当归是优化调控土壤环境,提高当归产出性能的有效轮作模式。     

The IMPDH inhibitors,ribavirin and mycophenolic acid,inhibit peste des petits ruminants virus infection

Peste des petits ruminants virus (PPRV) causes highly contagious diseases in domestic and particular wild small ruminants, leading to substantial economic loss. The development of effective and cheap antiviral medications shall help to circumvent this emerging burden. In this study, we found that ribavirin, a competitive inosine-5’-monophosphate dehydrogenase (IMPDH) inhibitor, significantly inhibits the replication of PPRV. As IMPDH is a key enzyme in purine nucleotide synthesis, supplementation of exogenous guanosine attenuate the anti-PPRV effect of ribavirin. Interestingly, an uncompetitive IMPDH inhibitor, mycophenolic acid (MPA), exerted more potent antiviral effect again PPRV. Similarly, this effect was largely restored upon supplementation of guanosine. Thus, we have demonstrated that the IMPDH inhibitors ribavirin and MPA combat PPRV infection through purine nucleotide depletion. Because both regimens have been widely used in the clinic for treating viral infection or organ rejection in transplantation patients for decades, respectively, repurposing these existing safe and cheap medications may provide a new avenue for combating PPRV infection.     

Effects of in ovo feeding of L‐arginine on the development of digestive organs,intestinal function and post‐hatch performance of broiler embryos and hatchlings

This study was to investigate the effects of in ovo feeding (IOF) L‐arginine (Arg) solution on the development of digestive organs, the duodenal mucosa of broiler embryos and hatchlings, and the growth performance of chicks during the first week post‐hatch. A total of 720 fertilized eggs with similar weight were randomly allocated to three groups, consisting of eight replicates of 30 eggs each. Three treatments were arranged as non‐injected control, diluent‐injected (0.75% NaCl solution) group and Arg solution‐injected group containing 1% Arg, dissolved in diluent. At 17.5 days of incubation, 0.6 ml of IOF solution was injected into amniotic fluid of each egg of injected groups. Results showed IOF of Arg solution increased (p < .05) the chick embryo weight at 19 days of incubation; the body weight gain of post‐hatch broilers during 1–7 days; the weights of liver, pancreas, proventriculus and gizzard; the concentrations of duodenal ghrelin, vasoactive intestinal peptide and glucagon‐like peptide 2; and the duodenum mucosal enzyme activities of alkaline phosphatase, maltase, sucrase and inducible nitric oxide synthase of 7‐day‐old post‐hatch broilers compared with other groups. The IOF of Arg solution also increased (p < .05) the villus height (VH) and the ratio of VH to crypt depth (CD) and decreased (p < .05) the CD in duodenum of broiler embryos and post‐hatch hatchlings, except for the CD at 19 days of incubation. In conclusion, IOF of 1% Arg solution into the amnion at 17.5 days of incubation could improve the development of digestive organs, the duodenal morphology, the releasing of gastrointestinal hormones and mucosal enzyme activities of broiler embryos and hatchlings and finally the growth performance of chicks during the first week post‐hatch. Therefore, IOF of appropriate Arg solution could be an effective technology for regulating early nutrition supply and subsequent growth development in poultry industry.     

Methyl donors dietary supplementation to gestating sows diet improves the growth rate of offspring and is associating with changes in expression and DNA methylation of insulin‐like growth factor‐1 gene

The study aimed to investigate the effects of maternal dietary methyl donors on the performance of sows and their offspring, and the associated hepatic insulin‐like growth factor‐1 (IGF‐1) expression of the offspring. A total of 24 multiparous sows were randomly fed the control (CON) or the CON diet supplemented with methyl donors (MD) at 3 g/kg betaine, 15 mg/kg folic acid, 400 mg/kg choline and 150 μg/kg VB₁₂, from mating until delivery. After farrowing, sows were fed a common lactation diet through a 28‐days lactation period and six litters per treatment were selected to be fed until at approximately 110 kg BW. Maternal MD supplementation resulted in greater birthweight (p < 0.05) and increased the piglet weights (p < 0.01) and litter weights (p < 0.05) at the age of day 28, compared with that in CON group. The offspring pigs in the MD group had greater ADG (p < 0.05) and tended to lower F:G ratio (p = 0.07) compared with that of CON group from day 28 to 180 of age. The offspring pigs from MD group had greater serum IGF‐1 concentrations and expressions of hepatic IGF‐1 gene and muscular IGF‐1 receptor (IGF‐1r) protein at birth (p < 0.05), and greater hepatic IGF‐1 protein (p = 0.03) and muscular IGF‐1r gene expressions (p < 0.05) at slaughter, than that from the CON group. Moreover, the methylation at the promoter of IGF‐1 gene in the liver of newborn piglets and finishing pigs was greater in the MD group than that of the CON group (p < 0.05). In conclusion, maternal MD supplementation throughout gestation could enhance the birthweight and postnatal growth rate of offspring, associated with an increased expression of the IGF‐1 gene and IGF‐1r, as well as the altered DNA methylation of IGF‐1 gene promotor.