马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

基于实时性优化的内燃机燃烧分析系统研究

采用缸压传感器、数据采集卡、光电编码器和Lab VIEW软件,搭建在线缸压采集和实时燃烧分析系统平台,研究HCCI和RCCI燃烧模式的循环波动特性。针对单循环缸压测量过程中的通道效应干扰,基于频谱分析,采用FFT、线性插值法和IFFT等方法相结合的滤波方式,实现共振峰的在线自适应识别与实时滤波,较好地减少了单循环缸压的干扰误差,使单循环的实时燃烧分析成为可能。随后,基于实时滤波后的缸压曲线,计算得到内燃机的最大压力升高率、燃烧放热率、爆发压力等重要燃烧参数。利用程序算法中同步性良好的生产者/消费者运算模式提高数据的共享能力,实现了数据实时运算与数据快速储存的并行处理,提高了燃烧计算的实时性;针对发动机不同工作阶段采用了不同精度层次的计算方法,减少了进排气、压缩和膨胀阶段的计算耗时,在计算量较大的燃烧放热率计算部分,通过适当简化计算公式和公式节点运算模块来提高燃烧系统的实时性。最后,分析了燃烧分析系统的实时性,并进行了燃烧分析系统的实验验证。     

乡土阔叶树种在碳汇造林中的应用及优势分析

随着我国社会经济的发展,逐步暴露出了许多的生态环境问题。而为了维持生态的平衡,就要提高植树造林的力度,这其中碳汇造林就是植树造林工作的延伸。在碳汇造林中乡土阔叶树种发挥着重要作用,针对乡土阔叶树种在碳汇造林中的应用进行分析,对项目中出现的问题进行优化。     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

基于形态学及多基因位点的枸杞褐斑病病原鉴定

枸杞Lycium barbarum Mill.是一种药食两用的名贵中药材,褐斑病是2010年在甘肃省枸杞种植区发生的由无性态类真菌新种枸杞小黑梨孢Stigmella lycii引起的新病害,据2015年-2018年调查,该病害已扩展至甘肃省各枸杞种植区,常年发病率45%～65%,严重度2～3级。本研究首次对枸杞小黑梨孢有性态形态进行描述,并结合分子生物学方法,确定该病病原菌的系统发育地位,为该病的防治提供一定的理论依据。枸杞褐斑病菌在离体培养条件下和越冬病叶上均可形成有性态。子囊壳球形至亚球形,大小185.6μm×176.6μm,有喙,喙的大小为(35.7～53.6)μm×(32.0～53.55)μm;子囊袋状,大小(103.2～165.7)μm×(15.7～22.4)μm,含8个子囊孢子;子囊孢子砖格状,大小(25.9～36.5)μm×(9.4～14.1)μm,平均31.2μm×12.3μm,具(4～)6～7个纵隔和1～3个横隔。通过ITS、LSU、RPB2和EF1-α多基因位点联合构建系统发育树,确定该菌为子囊菌门格孢腔菌目格孢腔菌科真菌。秋末冬初清洁田园,减少来年初侵染源,是有效防治枸杞褐斑病的关键措施。     

A comparison of the effects of carbon dioxide and medical air for abdominal insufflation on respiratory parameters in xylazine-sedated sheep undergoing laparoscopic artificial insemination

AIMS: To determine if abdominal insufflation with medical air will improve oxygenation and ventilation parameters when compared to insufflation with CO₂ in xylazine-sedated sheep undergoing laparoscopic artificial insemination (AI).

METHODS: Forty-seven sheep underwent oestrus synchronisation and were fasted for 24 hours prior to laparoscopic AI. Each animal was randomised to receive either CO₂ or medical air for abdominal insufflation. An auricular arterial catheter was placed and utilised for serial blood sampling. Respiratory rates (RR) and arterial blood samples were collected at baseline, after xylazine (0.1?mg/kg I/V) sedation, 2 minutes after Trendelenburg positioning, 5 minutes after abdominal insufflation, and 10 minutes after being returned to a standing position. Blood samples were collected in heparinised syringes, stored on ice, and analysed for arterial pH, partial pressure of arterial O₂ (PaO₂), and CO₂ (PaCO₂). The number of ewes conceiving to AI was also determined.

RESULTS: Repeated measures ANOVA demonstrated temporal effects on RR, PaO₂, PaCO₂ and arterial pH during the laparoscopic AI procedure (p<0.001), but no difference between insufflation groups (p>0.01). No sheep experienced hypercapnia (PaCO₂>50?mmHg) or acidaemia (pH<7.35). Hypoxaemia (PaO₂<70?mmHg) was diagnosed during the procedure in 14/22 (64%) ewes in the CO₂ group compared with 8/23 (35%) ewes in the medical air group (p=0.053). Overall, 15/20 (75%) ewes in the CO₂ group conceived to AI compared with 16/22 (72.7%) in the medical air group (p=0.867).

CONCLUSIONS AND CLINICAL RELEVANCE: There were no statistical or clinical differences in RR, PaO₂, PaCO₂, pH, or conception to AI when comparing the effects of CO₂ and medical air as abdominal insufflation gases. None of the sheep experienced hypercapnia or acidaemic, yet 42% (19/45) of sheep developed clinical hypoxaemia, with a higher percentage of ewes in the CO₂ group developing hypoxaemia than in the medical air group. Based on the overall analysis, medical air could be utilised as a comparable alternative for abdominal insufflation during laparoscopic AI procedures.     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

狗头枣GSTU基因的克隆及其序列分析

为探索红枣GSTU类基因在红枣抗逆中的分子作用机制,本研究以狗头枣枣树叶片的DNA为模板,根据枣GSTU基因的序列(HM345954.1)设计引物,采用PCR方法获得GSTU基因序列,并利用生物信息学手段对其所对应蛋白的结构域、功能域、理化特性、跨膜区、二级结构及亚细胞定位等进行分析。结果表明:克隆到的DNA序列全长838 bp,具有2个外显子和1个内含子,内含子在292~460 bp碱基处,该序列与GenBank中枣的GSTU核苷酸序列和氨基酸序列的同源性分别为99.85%和100%,表明成功的克隆了狗头枣的GSTU基因。该序列编码222个氨基酸,分子量为25.268 kD,理论等电点(pI)为6.10;亲水性的平均数为-0.242,表明该蛋白是亲水性蛋白。二级结构主要以α-螺旋为主,其次是无规则卷曲,不存在信号肽,表明该蛋白可能为非分泌性蛋白,该蛋白可能位于内质网(膜)和质膜上。本研究获得的狗头枣GSTU基因在基因结构上含有1个内含子,在核酸序列及氨基酸序列上与GenBank中枣的GSTU基因同源性高,并对其理化性质、结构等进行了预测与分析,为进一步研究红枣抗逆分子机制提供理论基础,同时为红枣品种的选育提供理论支持。     

米槠人工林土壤微生物群落组成对凋落物输入的响应

全球气候变化显著影响森林凋落物数量,进而会对土壤微生物群落造成影响。本研究以亚热带米槠人工林为研究对象,探究不同凋落物量输入处理(凋落物去除、凋落物加倍、对照)下,森林土壤微生物群落组成的变化。结果表明:与去除凋落物相比,凋落物加倍后0~10 cm土壤铵态氮(NH4^+-N)、硝态氮(NO3^--N)、全氮(TN)、有效磷(AP)含量分别显著增加了30.30%、49.66%、12.77%和13.90%。与对照相比,凋落物加倍与去除处理土壤微生物生物量碳(MBC)和氮(MBN)含量分别显著增加和下降(P<0.05),但凋落物加倍与去除处理间无显著差异。凋落物加倍处理下土壤丛枝菌根真菌(AMF)、革兰氏阳性菌[G(+)]、革兰氏阴性菌[G(-)]、放线菌(ACT)、真菌(F)丰度和总磷脂脂肪酸(TPLFA)含量分别比去除凋落物处理的土壤高68.35%、63.35%、82.65%、69.02%、40.56%和65.85%,而土壤革兰氏阳性菌与阴性菌比值、真菌与细菌比值则分别降低11.64%和26.67%。冗余度分析表明,铵态氮是影响该人工林土壤微生物群落组成的最主要环境因子。可见凋落物输入量变化改变了土壤养分有效性,进而显著影响了土壤微生物群落组成,这对进一步深入探究全球气候变化对亚热带森林土壤养分循环的影响具有重要意义。