Investigation of environmental and land use impacts in forested permafrost headwaters of the Selenga-Baikal river system,Mongolia - Effects on discharge,water quality and macroinvertebrate diversity

Headwater streams play a major role for provision of ecosystem services, e.g. drinking water. We investigated a high-altitude headwater catchment of the Kharaa River (including 41 1st-order rivers) to understand the impact of land cover (especially forest cover), environment and human usage on runoff, chemical water quality and macroinvertebrate fauna in a river basin under discontinuous permafrost conditions in an arid, sparsely populated region of Mongolia. To verify our hypotheses that different landuses and environmental impacts in permafrost headwaters influence water quality, we investigated 105 sampling sites, 37 of them at intermittent stream sections without water flow. Discharge was positively impacted by land cover types steppe, grassland and forest and negatively by shrubland, forest burnt by wild fires (indicating a reduction of permafrost) and slope. Water quality was affected by altitude, longitude and latitude, shrub growth and water temperature. Shannon diversity of macroinvertebrates was driven by water temperature, iron content of the water, flow velocity, and subbasin size (adjusted R² = 0.54). Sample plots clustered in three groups that differed in water chemistry, macroinvertebrate diversity, species composition and bio-indicators. Our study confirms that steppes and grasslands have a higher contribution to runoff than forests, forest cover has a positive impact on water quality, and diversity of macroinvertebrates is higher in sites with less nutrients and pollutants. The excellent ecological status of the upper reaches of the Kharaa is severely threatened by forest fires and human-induced climate change and urgently needs to be conserved.     

绵羊心脏脂肪酸结合蛋白基因单核苷酸多态性分析

试验旨在探讨心脏脂肪酸结合蛋白（heart fatty acid-binding protein，H-FABP）基因在绵羊中的遗传多态性，并寻找可用于辅助选择的分子标记。本研究以滩羊（250只）及滩羊×湖羊杂交F1代（174只）为试验动物，利用SNaPshot分型技术对H-FABP基因（GenBank登录号：AY157617）的多态位点进行单核苷酸多态性（SNP）分析，统计基因频率和基因型频率，进行Hardy-Weinberg平衡性检测，计算期望杂合度（He）、多态信息含量（PIC）和有效等位基因数（Ne）等遗传多态指标，分析候选基因不同基因型与体重、体长、体高、胸围、胸深、胸宽和管围等生长性状的关联性。结果显示：①检测到9个多态位点：939[A/G]、980[G/A]、1018[T/C]、2878[C/T]、2956[C/T]、3017[G/A]、3341[G/C]、3394[T/A]、1056[-/G]，其中有6处转换、2处颠换、1处单碱基插入。②939[A/G]、980[G/A]、2956[C/T]、3341[G/C]、3394[T/A]和1018[T/C]的He为0.3200~0.4666，PIC为0.2688~0.3577；2878[C/T]、3017[G/A]位点的He为0.0283~0.1272，PIC为0.0279~0.1191，为低度多态；1056[-/G]位点的He在滩羊、滩羊×湖羊杂交F1代群体中分别为0.0120和0，PIC在滩羊、滩羊×湖羊杂交F1代群体中分别为0.0119和0；9个多态位点在滩羊、滩羊×湖羊杂交F1代群体中均符合Hardy-Weinberg平衡定律。③5个多态位点：939[A/G]、980[G/A]、2956[C/T]、3341[G/C]、3394[T/A]处于紧密连锁（D'>0.99），将H-FABP基因分为3个单倍型：AA、AB和BB。④在41只滩羊×湖羊杂交F1代羊中，BB单倍型在各生产性状上具有最大值，但各单倍型间差异不显著（P>0.05）。结果表明，绵羊H-FABP基因具有丰富的遗传多态性，939[A/G]、980[G/A]、2956[C/T]、3341[G/C]、3394[T/A]5个位点紧密连锁，BB单倍型可能是与绵羊生产性状相关的优势单倍型。     

Construction and Partial Biological Characteristics Analysis of Listeriolysin S llsB Gene Deletion Strain

In order to analyze the effect of listeriolysin S (LLS) llsB gene deletion on the biological characteristics of Listeria monocytogenes (LM),this study used homologous recombination to construct the llsB gene deletion strain LM90-ΔllsB,and the biological characteristics of growth characteristics,median lethal dose (LD₅₀) and organ-borne bacteria were studied in healthy Kunming male mice at 8 weeks old and weighing 40 g±5 g.The llsB gene deletion strain was successfully constructed,and the deletion strain had good genetic stability through continuous passage to 20 generations in vitro.Based on its growth curve examination,we found that the growth rate of the mutant strain was slightly higher than that of the parent strain.The results of mice infection test showed that the LD₅₀ of the parent strain and the deletion strain were 10^6.17 and 10^6.50 CFU,respectively.Compared with the parent strain,the amount of bacteria load of the deletion strain in the liver and spleen of the mice was extremely significantly decreased (P<0.01),the results showed that the infection ability of the mutant strain on mice was obviously weakened.No Listeria monocytogenes was detected in the brain.The results suggested that llsB gene might have direct or indirect regulatory effect on some biological characteristics of LM90,and it would provide a theoretical basis for further understanding of the pathogenic mechanism of LLS,prevention and control of listeriosis.     

Intestinal Microbiota in Large Yellow Croaker,Larimichthys crocea,at Different Ages

The polymerase chain reaction–denaturing gradient gel electrophoresis (PCR‐DGGE) of 16S ribosomal RNA gene was used to investigate bacterial communities in the intestines of large yellow croaker at six different ages (12 d, 18 d, 26 d, 40 d, 3 mo, and 1 yr old) as well as within the corresponding feed and culture water. In addition, Illumina Miseq sequencing was utilized to compare intestinal microbiota between 12‐d‐old and 1‐yr‐old individuals. PCR‐DGGE results revealed that the culture water had the highest bacterial diversity, followed by the feed, while the intestines had the lowest diversity. The intestinal microbiota at six ages changed severely; however, the change did not follow any trend. The large yellow croaker intestines harbored specific bacterial communities that differed from those in both feed and water. Illumina Miseq sequencing results revealed that the diversity of intestinal bacteria in 12‐d‐old fish was higher than that in 1‐yr‐old fish, and the bacterial composition differed significantly between them. γ‐Proteobacteria and Pseudoalteromonas supplied the most abundant phylum and genus in the 12‐d‐old fish intestine. However, in the 1‐yr‐old fish intestine, Firmicutes and Clostridium were the most dominant, respectively. The study may contribute to a better understanding of gut microbiota and dynamics of the large yellow croaker and the relationship with their surrounding environment.     

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。     

表达猪乳铁蛋白肽重组屎肠球菌对断奶仔猪生长性能的影响及其抗ETEC感染效果研究

本试验以表达猪乳铁蛋白肽的重组屎肠球菌（pNZ8112-PLFcin/Ef）饲喂断奶仔猪，研究其对仔猪生长性能的影响和抗产肠毒素大肠杆菌（enterotoxigenic Escherichia coli，ETEC）感染的效果。选取28日龄体重相近的健康断奶仔猪36头，随机分为3组（重组屎肠球菌组、空载体组和培养基组），每组3个重复，每个重复4头仔猪。重组屎肠球菌组和空载体组分别饲喂添加pNZ8112-PLFcin/Ef （6×10¹² CFU/kg）和pNZ8112/Ef （6×10¹² CFU/kg）的基础日粮，而培养基组饲喂含有相同体积的GM17液体培养基的基础日粮。试验期26 d。结果显示，与培养基组相比，重组屎肠球菌组断奶仔猪的平均日增重极显著提高（P<0.01）；料重比显著降低（P<0.05）；腹泻率明显降低，肠道菌群的均匀度和多样性指数均下降。为进一步探究表达猪乳铁蛋白肽的重组屎肠球菌对断奶仔猪抵抗ETEC感染的保护作用，在连续饲喂21 d后，每个重复中随机挑选1头体况相近的断奶仔猪灌服ETEC。结果发现，与培养基组相比，攻菌后重组屎肠球菌组断奶仔猪血清白细胞介素-2（IL-2）、免疫球蛋白G （IgG）含量、肠黏液中分泌型免疫球蛋白A （sIgA）水平均显著升高（P<0.05）；脾脏指数显著提高（P<0.05），但胸腺指数、肠段长度及重量则均无显著差异（P>0.05）。综上所述，表达猪乳铁蛋白肽的重组屎肠球菌能够起到促进断奶仔猪生长及抗ETEC感染的保护作用。     

Application of molybdenum fertilizer enhanced quality and production of alfalfa in northern China under non-irrigated conditions

Molybdenum (Mo) is an essential micro-mineral nutrient for plant growth and development. As an important legume forage, alfalfa (Medicago sativa) is frequently planted in marginal lands of northern China where water scarcity and infertile soil are limiting factors for alfalfa production. Mo plays an important role in nitrogen (N) metabolism in general and N fixation in legume species in particular. To evaluate the role of Mo fertilizer in alfalfa quality and production in semi-arid areas of northern China, we conducted a three-year field experiment in Inner Mongolia from 2012 to 2014. Mo fertilizer at the doses of 0, 150 g hm^?2, 300 g hm^?2yr^?1 was applied consecutively across the three years. Application of Mo at 300 g hm^?2yr^?1 significantly increased dry yield of alfalfa in 2014. Crude protein (CP) content of alfalfa without application of Mo exhibited a decline trend from 2012 to 2014. Application of Mo significantly enhanced total CP in 2012 and 2014, and delayed the reduction in total CP across the three years. In contrast to CP, Mo fertilizer had no significant effects on neutral detergent fiber and acid detergent fiber content. In addition, we found that nitrate reductase activity of alfalfa was increased by Mo fertilizer, which may account for the enhanced CP content in alfalfa. These findings highlight that application of Mo fertilizer can be an effective way to improve alfalfa production and quality in semi-arid areas in northern China.     

柑橘褐斑病菌对不同抗性柑橘品种防御酶活性的影响

以柑橘褐斑病高抗品种Lw8,中抗品种L2,高感品种Lw14为实验材料,通过孢子喷雾法使柑橘褐斑病菌感染柑橘叶片,研究接菌0d、1d、2d、3d、6d、8d、10d时,柑橘叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、几丁质酶(CHT)酶和β-1,3葡聚糖酶(GLU)含量及活性变化规律与品种抗病性的关系。结果表明,接种前,两个抗病品种Lw8、L2中的POD、SOD、PAL、PPO 、GLU、CHT活性显著高于高感品种Lw14,CAT活性则与高感品种Lw14相近；接种后,3个不同抗性品种中SOD、POD、CAT、PAL、PPO 、GLU、CHT 7种防御酶活性较对照明显提高,但不同品种增加幅度不一致,除CAT外,其余6种防御酶活性均表现为抗病品种Lw8、L2增幅显著高于高感品种Lw14,且两个抗病品种一般在接菌3d内6种防御酶活性迅速升高并达到峰值；而高感品种Lw14防御酶(除CAT外)活性或增幅较小或较两个抗病品种滞后。本研究初步探讨了不同抗性品种接种褐斑病菌后7种防御酶的活性动态变化与柑橘品种抗病性关系,为进一步研究其抗病机理奠定基础。     

Comparative proteomics analysis of maize (Zea mays) leaves infected by small brown planthopper (Laodelphax striatellus)

Maize rough dwarf disease (MRDD) is a viral disease caused by brown planthopper infestation, and leads to great yield loss, especially in China. Comparative proteomics was performed using maize inbred line Zheng 58 and LN 287. MRDD pathogen was detected as rice black-streaked dwarf virus (RBSDV) by quantitative real time PCR (qRT-PCR) in Shandong Province, China. The modified trichloroacetic acid (TCA)/acetone method was used for soluble protein extraction from leaves. Two-dimensional electrophoresis (2-DE) analysis was performed on 24-cm long, pH 4–7 linear immobilized pH gradient (IPG) strips, and gels were stained with silver and coomassie brilliant blue. We identified 944 proteins expressed in RBSDV infected maize leaves by proteomics approaches. Among these, 44 protein spots that revealed a 1.5-fold difference in intensity were identified by mass spectrometry between mock-inoculated and RBSDV infected samples. Among these, 17 and 26 spots were up-regulated, and 27 and 18 spots were down-regulated in the virus infected samples of Zheng 58 and LN 287, respectively. Differential protein spots were analyzed by mass spectrometry identification, which could be divided into six categories. Furthermore, the expression of stress-related proteins was detected and confirmed by qRT-PCR. This study lays the foundation for further investigations, enabling the enhancement of MRDD resistance in maize.     

Production of Antioxidative Maillard Reaction Products from Gelatin Hydrolysate of Unicorn Leatherjacket Skin

Antioxidative activities of Maillard reaction products (MRPs) derived from gelatin hydrolysate (GH) of unicorn leatherjacket skin as affected by types of saccharides, GH to saccharide ratio, incubation temperatures, relative humidity (RH), and times were investigated. MRPs prepared using mixture of GH and galactose at 2:1 (w/w) ratio showed the highest antioxidative activity. At different incubation temperatures (50–70°C) and RHs (55–75%), optimal condition for preparing antioxidative MRPs included heating GH and galactose (2:1) powder at 70°C and 55% RH for 36 h. Electrophoresis and Fourier transform infrared spectroscopy revealed the formation of high molecular weight polymers, and the advanced glycation took place in MRPs.