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.     

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。     

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.     

杂草环境下转EPSPS基因大豆NZL06-698的生态适应性研究

为研究转EPSPS基因大豆NZL06-698在杂草环境中的生态适应性,试验设置杂草处理模拟野外环境,研究转基因大豆NZL06-698(GT698)的生存竞争能力以及外源EPSPS蛋白表达情况。结果表明:在相同处理下转基因大豆GT698的株高(R8期)、百粒重显著高于亲本大豆蒙豆12(MD12),但单株产量、单株籽粒数、结实率显著低于亲本大豆,说明外源基因的存在并未增强转基因大豆的生存竞争力,且转基因大豆在野外的生存竞争能力与亲本大豆相比较弱。试验注意到,杂草环境对转EPSPS基因大豆的生长有明显的限制作用,杂草处理下转基因大豆的单株产量、单株籽粒数、百粒重、结实率等指标均显著低于对照处理。ELISA检测结果表明,转基因大豆叶片及籽粒中外源EPSPS蛋白在杂草处理及对照中均正常表达,且两种处理下的EPSPS蛋白表达量无显著差异。以上结果表明杂草环境中转基因大豆的EPSPS蛋白正常表达,正常提供抗草甘膦性状,但是外源基因的存在并没有增加转基因大豆的生存竞争能力,且转基因大豆的生长受到杂草环境的显著抑制,由此得出结论:与亲本大豆相比,转EPSPS基因大豆NZL06-698在野外环境中生态适应能力较弱。     

草坪草屑的加工与利用

我国每年修剪草坪产生大量的草坪草屑,对草坪草屑的加工利用,可以对我国能源、饲料、肥料及蛋白资源的短缺起到一定的缓解作用,因此如何对草坪草屑进行合理利用已成为一项重要的研究课题。本文综述了草坪草屑能源化、饲料化、肥料化和深加工4种利用方式,详细介绍了草坪草屑在发酵生产能源、饲喂家畜、堆肥还坪以及加工叶蛋白等方面的研究进展,指出禾本科草坪草屑和豆科草坪草屑应采用不同的处理方式,对各加工方式的投入和产出也应进行进一步的量化核算,同时总结并展望了草坪草屑利用方式的发展前景。     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.