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。     

厚壳贻贝5-羟色胺2A受体(5-HT2AR)基因克隆和时空表达

为探究5-羟色胺2A受体（5-HT 2A receptor， 5-HT2AR）基因在海水贝类生长发育中的作用，本研究通过RACE技术和实时荧光定量PCR（qRT-PCR）克隆了厚壳贻贝5-HT2AR 基因的cDNA全长，并分析该基因的时空表达。结果显示，5-HT2AR基因全长2 636 bp，开放阅读框（ORF）2 124 bp，共编码707个氨基酸。序列分析结果显示该序列与人、小鼠、斑马鱼、长牡蛎和虾夷扇贝等物种分别具有45%、45%、48%、49%和67%的同源性。厚壳贻贝雌雄成体各组织和器官中均有5-HT2AR基因表达，雄性中的鳃表达量最高，而在雌性鳃、外套膜和性腺中的表达稍高于其他组织和器官；推测该基因可能与厚壳贻贝的摄食、对外界环境的感知及促进卵母细胞成熟有关。5-HT2AR基因在厚壳贻贝各发育阶段均有表达，且稚贝的表达量为眼点幼虫的1.4倍，推测5-HT2AR基因可能参与了调控厚壳贻贝幼虫的生长发育过程。本研究为进一步了解5-HT基因家族在双壳贝类中的功能奠定了基础。     

Bamboo NiR gene is associated with regeneration capacity

It was reported that the activity of NiR (nitrite reductase) enzyme encoded by the NiR gene was correlated with the regeneration ability in rice. To testify the function of NiR gene in bamboo, seven bamboo species whose calli had different differentiation rate were chosen to analyse their NiR enzyme activity. The results showed that bamboo NiR enzymatic activity had a certain correlation with the regeneration capacity. A NiR gene named DhNiR from Dendrocalamus hamiltonii with high regeneration capacity was cloned. Sequence analysis revealed that the ORF of DhNiR was 1779bp encoding 592 amino acids. Overexpression of DhNiR in rice reduced the time of shoot differentiation and increased the transformation rate, suggesting that DhNiR might play an important role in the regeneration ability of bamboo, and can be applied in regeneration and gene transformation of bamboo and other plant species.     

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。     

Aquaponics and sustainability: The comparison of two different aquaponic techniques using the Life Cycle Assessment (LCA)

Aquaponics is generally regarded as a sustainable practice, but its environmental burdens were not yet deeply investigated. In this study, Life Cycle Assessment (LCA) was used to assess the environmental impacts of two hypothetical coupled aquaponics systems (CAPS): Raft System (RAFT) and Media-Filled Beds System (MFBS). Rainbow trout (Oncorhynchus mykiss) and lettuce (Lactuca sativa) were considered as cultivated species in both systems. The Simapro^© software V.8.0 was used for calculation. The comparison between the two virtual systems indicated the floating technique as the less impacting one. Even though energy consumption appears to be higher in the floating system, LCA results were markedly influenced by the extensive use of inert materials in MFSB. In both systems, contribution analyses underlined that the main environmental impacts are related to infrastructures, electricity and fish feed. The LCA analyses carried out in this study highlights that the choice of less impacting materials, and the optimization of management practices, should be taken as priorities in order to reduce environmental impacts of this activity.     

Immunostimulatory effects of dietary poly‐β‐hydroxybutyrate in European sea bass postlarvae

The stable production of high‐quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly‐β‐hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In this study, the effect of feeding PHB‐enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) postlarvae was examined. Amorphous PHB was administered to 28‐day‐old sea bass postlarvae over a period of 10 days. The survival and growth performance were monitored, and the expression of 29 genes involved in immunity, growth, metabolism and stress‐response was measured. While the expression of the insulin‐like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed postlarvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies.     

Molecular cloning and functional characterization of the fatty acid delta 6 desaturase (FAD6) gene in the sea cucumber Apostichopus japonicus

The sea cucumber (Apostichopus japonicus), an important echinodermata, had high value in nutrition and medicine for its rich collagen, sulphated polysaccharide, glycosides and polyunsaturated fatty acids (PUFA). The cDNA of the fatty acid desaturase gene in A. japonicus (AJFAD6) was cloned and was found to encode a desaturase with delta 6 FAD activity. Sequence analysis indicated that AJFAD6 included an open reading frame of 1392 bp, encoding 463 amino acids. AJFAD6 has all the conserved motifs found in other members of the FAD6 family, including an N‐terminal cytochrome b5 domain and three histidine‐rich regions. qRT‐PCR showed that AJFAD6 was expressed in all tissues tested during juvenile development and was mainly expressed in the respiratory tree at 150 days after adherence (150 days) and in the intestine at 100 days. Furthermore, AJFAD6 mRNA was also detected in the analysed adult tissues, with higher expression in the intestine and testis. Functional characterization of AJFAD6 in a recombinant yeast, Pichia pastoris, showed that AJFAD6 could catalyse exogenous linoleic acid (LA) and α‐linolenic acid (ALA) to produce γ‐linoleic acid (GLA) and stearidonic acid (STA), respectively, at conversion rates of 11.1% for LA to GLA and 3.4% for ALA to STA. Our results suggested that the biosynthetic pathway of PUFA existed in the sea cucumber, but endogenous production of eicosapentaenoic acid, arachidonic acid and docosahexaenoic acid from either LA or ALA precursor appeared to be limited.     

射流式鱼泵输送草鱼的性能研究

为了分析射流式鱼泵输送草鱼的性能及其损伤因素,文章设计了一台喉管直径为60 mm的射流式鱼泵,开展了草鱼输送实验,并采用高速摄影和计算流体力学方法进行了研究。结果显示,该射流式鱼泵在扬程2.24m时最高草鱼输送能力达918 kg·h~(-1),其所需水功率为2.83 k W。进一步的检测表明,部分实验鱼有鳞片脱落的情况,但未出现游泳异常,解剖后也未发现内脏受损等情况;实验鱼在过泵后呼吸频率及部分血液指标存在明显变化,但在24 h内基本可以恢复。数值模拟和高速摄影方法分析得出,剪切层是造成实验鱼泵内鳞片脱落的主要原因,撞击伤是由内流偏转诱导实验鱼撞击泵内壁面产生的,包含压力梯度在内的水力因素都可能使实验鱼产生应激反应。但由于鱼类在泵内时间极短,上述因素都不会致实验鱼死亡。