Presence of Muscarinic Acetylcholine Receptors in the Cattle Tick Boophilus microplus and in Epithelial Tissue Culture Cells of Chironomus tentans

A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [³H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM ; B_max 22·5 pmol mg protein⁻¹) were detected that do not bind nicotinic compounds specifically. The estimated IC₅₀ values for nicotine, imidacloprid and α-bungarotoxin were all in the mM range. Additionally, with tick larvae, high-affinity nicotinic binding sites were detected with [³H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC₅₀ values for nicotine, α-bungarotoxin, imidacloprid and QNB were 43(±8) nM , 0·8(±0·2) μM , 2·8(±0·6) μM and 78(±1·9) μM , respectively. With homogenates of the non-neuronal insect cell line from C. tentans, only high-affinity binding sites for [³H]QNB were found. Muscarinic antagonists selectively displaced [³H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC₅₀ 2·13(±1·02) μM ) among different subtype-specific mAChR antagonists (4-DAMP had IC₅₀ 49·9(±9·13) μM and methoctramine had IC₅₀ 121(±14·2) μM ) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G-protein-coupled receptor, as concluded from the 4·8-fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non-hydrolysable GTP-analogue γ-S-GTP. Binding data for the agonists oxotremorine M (IC₅₀ 71·3(±19·6) μM ) and carbachol (IC₅₀ 253(±87·1) μM ) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.     

基于网格单元的乌鲁木齐市土地覆被/利用时空变化

随着城市化进程在全球范围内的快速推进,对城市区域土地覆被/土地利用变化产生影响。采用1990、2000和2011年3期Landsat TM5遥感数据,提取乌鲁木齐市土地覆被/土地利用信息,并将各期用地分类数据分别转入250 m×250 m的矢量网格单元,计算每个网格单元内各类用地面积占网格单元面积比例的变化,探讨乌鲁木齐市1990—2011年土地覆被/土地利用时空变化特征及驱动因素,并通过相关和回归分析,研究1990—2011年各用地类型之间相互转移和替代的关系。结果表明:1)1990—2011年乌鲁木齐市城市快速扩展,大面积裸地和绿地转化为建设用地,建设用地持续增加,从1990年157.37 km~2增加到2011年的444.89 km~2,裸地持续减少,绿地先减少后增加,水体则呈现缓慢增长的趋势。2)研究区内不同区域和方向由于受到地形条件、政策、城市规划以及道路交通网络等因素的影响,土地覆被/土地利用呈现出不同的时空变化趋势,建成区老城区内部呈现挖潜的集约式增长,外围则是蔓延式扩展。该研究对该地区城市建设和发展具有借鉴意义。     

8种野生葱属植物的叶形态特征及其分类学意义研究

【目的】比较8种野生葱属植物的叶形态特征,为其分类学研究和生态适应性机制分析提供依据。【方法】以分布于新疆不同生境的8种野生葱属植物齿丝山韭、实葶葱、新疆蒜、健蒜、多籽蒜、奇台蒜、喀纳斯蒜和乌鲁木齐蒜为研究对象,采用田间观测和树胶制片法,测定其叶片数量、长度、宽度等指标,观察其解剖结构异同;通过相关性分析和聚类分析,研究叶片表皮性状之间的关系以及性状的分类学意义。【结果】8种葱属植物的叶形态和微形态存在差异,实葶葱和齿丝山韭叶直立,其余呈不同程度倒伏;实葶葱叶圆柱形,其余叶均为条形;实葶葱叶表面被蜡粉,其余均为光滑;实葶葱叶表皮细胞矩形,其余均为菱形;实葶葱垂周壁直线型,其余均为弧线形。8种植物叶片均为双面气孔型,气孔椭圆形,保卫细胞肾形,无副卫细胞,保卫细胞不同程度下陷。基于叶表皮结构性状的聚类分析结果显示,供试8种材料分为3类,齿丝山韭和实葶葱分别聚为一类,其余6种聚为一类。【结论】叶形态、表皮细胞形状、垂周壁形状、表皮细胞密度和气孔指数大小可为种的界定提供重要的形态分类学依据,具有分类学意义。叶片上表皮细胞比下表皮细胞可以提供更为全面的微形态特征。新疆8种野生葱属植物的叶片形态和微形态结构均呈现一定的抗旱和抗寒特征。     

蝉花宝牌蝉花片免疫功能评价

将蝉花宝牌蝉花片(商品名)设低、中、高3个剂量组,分别相当人体推荐量的5、10、30倍,同时设阴性对照组(1%羧甲基纤维素钠),连续1个月经口给予小鼠不同剂量的样品,进行迟发型超敏反应(DTH)、腹腔巨噬细胞吞噬鸡红细胞、抗体生成细胞检测、自然杀伤(NK)细胞杀伤活性和脾淋巴细胞转化等实验,以评估其免疫功能。结果表明,与阴性对照组比较,受试3个剂量组足跖肿胀度极显著增加(P0.01);中、高剂量组溶血空斑数和血清溶血素抗体积数值均极显著增加(P0.01),腹腔巨噬细胞吞噬鸡红细胞的吞噬率、吞噬指数显著增加(P0.05,P0.01);高剂量组NK细胞活性显著升高(P0.05)。蝉花宝牌蝉花片(商品名)具有显著的免疫调节功能。     

不同年限设施菜地番茄细胞壁果胶Cd累积的研究

为探讨不同种植年限和设施菜地番茄细胞壁果胶Cd累积的关系,以种植年限为2年和14年设施菜地番茄为研究对象,测定了不同种植年限下番茄茎、叶细胞壁果胶含量,并对细胞壁果胶含量、果胶甲酯酶(PME)活性与种植年限及Cd累积量之间的相关性进行了分析。结果表明,随着种植年限的增加,番茄植株各器官Cd累积量均有所增加;种植14年设施菜地番茄茎中Cd累积量比种植2年的增加了24.55%,叶片中Cd累积量增加了45.96%;种植14年设施菜地番茄茎细胞壁Cd累积量增加了61.86%,叶片细胞壁Cd累积量增加了57.46%;不同种植年限设施菜地番茄茎和叶细胞壁果胶Cd含量也存在极显著差异(P0.05)。另外,随着种植年限的增加,番茄茎、叶细胞壁果胶含量和PME活性随之升高:与种植2年相比,种植14年设施菜地番茄茎中果胶含量提高了104.51%,叶片果胶含量提高了127.45%;种植14年设施菜地番茄茎中PME活性提高了10.56%,叶片PME活性提高了9.39%。果胶含量及PME活性和细胞壁Cd累积量呈正相关。研究发现不同年限设施菜地土壤全Cd和有效Cd含量有所不同,影响着番茄不同器官果胶含量及PME活性,说明细胞壁果胶在缓解番茄Cd毒害中起着一定的作用。     

枣果皮细胞壁代谢酶活性与抗裂果的关系

【目的】测定枣果皮细胞壁代谢酶活性,研究果皮细胞壁代谢酶活性与抗裂果的关系。【方法】以抗裂枣品种扁核酸与易裂枣品种京枣39为研究对象,测定生长过程中枣果皮不同部位细胞壁代谢酶活性,比较抗裂和易裂枣品种果皮细胞壁代谢酶活性的差异。【结果】抗裂枣品种不同时期不同部位果皮的超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、多酚氧化酶(PPO)活性显著或极显著高于易裂枣品种,而在绿熟期不同部位抗裂品种的过氧化氢酶(CAT)活性极显著低于易裂品种。【结论】易裂品种果皮果肩阴面POD活性和PPO活性在白熟期显著高于抗裂品种,而其余时期其余部位则是抗裂品种的POD活性和PPO活性显著高于易裂品种,易裂品种果皮的梗洼、果顶、果肩阳面和果肩阴面CAT活性在绿熟期极显著高于抗裂品种,而其余时期则是抗裂品种高于易裂品种。     

高粱幼叶细胞悬浮系的建立

以高粱无菌苗幼叶为试验材料,对颗粒状胚性愈伤组织的获得、细胞悬浮系的建立及影响悬浮细胞生长的主要因素进行了研究。结果表明：幼叶在MS（Murashige and Skoog）+2mg/L 2,4-D培养基上诱导出的愈伤组织,经2~3次继代筛选后获得了浅黄色、颗粒状胚性愈伤组织;胚性愈伤组织接种于液体培养基中,于25±1℃、黑暗条件下,经45~60d的悬浮震荡培养建立了高质量的细胞悬浮系,细胞生长曲线呈“S”型,细胞密度可达6.45×10 ⁵~5.08×10 ⁶个/mL以上,活细胞率可达72.76%,近圆细胞率可达87.50%;培养基的基本成分、激素种类和水平对悬浮细胞生长状态有很大的影响,相比1/2MS培养基,MS培养基为适宜的培养基,2,4-D对保持细胞系的稳定增殖至关重要,适宜的浓度为1mg/L;培养液中加入0.5mg/L KT或6-BA会造成悬浮液褐化,影响悬浮细胞的生长;最适宜的细胞悬浮培养基为L2培养基（MS+1mg/L 2,4-D,30g/L蔗糖,pH5.8）,震荡培养转速为110~120r/min,继代周期为7d,新旧培养基的接种比例为2∶1。     

马铃薯干腐病病原镰孢菌体内产细胞壁降解酶特性研究

干腐病是马铃薯窖储期的主要病害,病原菌往往通过薯块表面的机械擦伤入侵块茎并破坏块茎的薄壁细胞而进行侵染,其中病菌的进一步扩展需要降解寄主细胞壁,因此,针对干腐病病原镰孢菌产酶特性的研究具有重要意义。以接骨木镰孢菌（Fusarium sambucinum）、燕麦镰孢菌（Fusarium avenaceum）、拟丝孢镰孢菌（Fusarium trichothecioides）、黄色镰孢菌（Fusarium culmorum）、拟枝孢镰孢菌（Fusarium sporotriodides）为试验菌种,以克新1号为寄主材料,对5种镰孢菌体内产细胞壁降解酶特性及温度对这些致病菌的产酶特性的影响进行了研究。结果表明,当5种镰孢菌接种在马铃薯薯块上后,均可以产生羧甲基纤维素酶（Cx）、β-葡萄糖苷酶、多聚半乳糖醛酸酶（PG）、果胶甲基半乳糖醛酸酶（PMG）,其中Cx的活性在侵染后期最高,PMG在侵染前期较高,且Cx活性低于PMG。在温度对镰孢菌活体内产细胞壁降解酶的影响试验中发现,相对于不接菌对照,Cx、PMG的活性在25℃时较高,β-葡萄糖苷酶在20℃下活性较高,PG在20℃和25℃的活性都较高。以上结果说明：镰孢菌侵染马铃薯时会产生大量的细胞壁降解酶,Cx活性在侵染后期较高,PMG活性在侵染前期较高,且低温不利于真菌细胞壁降解酶的产生。     

Development of a continuous cell line from larval Atlantic cod (Gadus morhua) and its use in the study of the microsporidian,Loma morhua

In vitro cell culture methods are crucial for the isolation, purification and mass propagation of intracellular pathogens of aquatic organisms. Cell culture infection models can yield insights into infection mechanisms, aid in developing methods for disease mitigation and prevention, and inform commercial‐scale cultivation approaches. This study details the establishment of a larval cell line (GML‐5) from the Atlantic cod (Gadus morhua) and its use in the study of microsporidia. GML‐5 has survived over 100 passages in 8 years of culture. The line remains active and viable between 8 and 21°C in Leibovitz‐15 (L‐15) media with 10% foetal bovine serum and exhibits a myofibroblast phenotype as indicated by immuno‐positive results for vimentin, α‐smooth muscle actin, collagen I and S‐100 proteins, while being desmin‐negative. GML‐5 supports the infection and development of two microsporidian parasites, an opportunistic generalist (Anncaliia algerae) and cod‐specific Loma morhua. Using GML‐5, spore germination and proliferation of L. morhua was found to require exposure to basic pH and cool incubation temperatures (8°C), in contrast to A. algerae, which required no cultural modifications. Loma morhua‐associated xenoma‐like structures were observed 2 weeks postexposure. This in vitro infection model may serve as a valuable tool for cod parasitology and aquaculture research.     

Partial replacement of soybean meal with Methylobacterium extorquens single‐cell protein in feeds for rainbow trout (Oncorhynchus mykiss Walbaum)

A feeding trial was conducted with juvenile rainbow trout (15–16 g initial weight) to assess the effects of including single‐cell protein (SCP) produced from Methylobacterium extorquens in trout feeds. Three isonitrogenous and isoenergetic diets were produced: a control diet and two experimental diets containing 5% or 10% bacterial protein meal replacing soybean meal. Triplicate tanks, each containing 35 fish, were fed each diet to apparent satiation in a constant‐temperature (15°C), flow‐through tank system for 12 weeks. No statistically significant differences in final fish weight or other fish growth parameters were observed. Similarly, feed efficiency parameters showed no significant differences among groups. Nutrient retention indices (protein, fat, energy) were relatively high and similar among fish in each dietary treatment group, as were whole body proximate compositions. Fish survival was high, with a small but statistically significant increase for the 10% SCP diet. Overall, results demonstrate that SCP from M. extorquens is a safe and effective alternative protein for rainbow trout diets at the low inclusion levels tested. Slightly lower weight gain in fish fed the 10% SCP diet was largely due to lower feed intake, suggesting that adding palatability‐enhancing ingredients to feeds may allow higher levels of M. extorquens SCP to be used without compromising fish growth.