柔嫩艾美耳球虫含HD结构域蛋白特性和功能初步研究

鸡球虫病是由艾美耳球虫引起的一种危害严重的肠道寄生虫病,每年都会给世界各地的养禽业带来巨大的经济损失。目前,该病主要依靠抗球虫药物进行防治,但由于药物的长期及不合理使用导致鸡球虫几乎对所有使用过的抗球虫药均产生耐药性。为研究球虫耐药性产生的分子机制,本实验室前期对柔嫩艾美耳球虫地克珠利耐药株、马杜拉霉素耐药株以及敏感株进行了转录组测序并获得了敏感株与耐药株的差异表达基因,发现柔嫩艾美耳球虫含HD域蛋白（EtHDCP）在耐药株中上调表达。本研究以柔嫩艾美耳球虫敏感株孢子化卵囊cDNA第一链为模板,成功克隆出EtHDCP基因,构建了原核表达重组质粒pGEX-4T-EtHDCP,并成功诱导表达了重组蛋白rEtHDCP。利用qRT-PCR和Western blot对柔嫩艾美耳球虫敏感株不同发育阶段的转录和翻译水平进行分析,结果显示,EtHDCP在第二代裂殖子的转录和翻译水平高于其他三个阶段（未孢子化卵囊、孢子化卵囊和子孢子）。同时利用Western blot分析了EtHDCP在柔嫩艾美耳球虫敏感株、地克珠利耐药株、马杜拉霉素耐药株中的翻译水平,结果显示,EtHDCP在耐药株中的蛋白翻译水平显著高于敏感株。间接免疫荧光定位结果显示,该蛋白主要定位在子孢子和裂殖子的表面以及裂殖子的胞质内。入侵抑制试验表明,抗rEtHDCP多克隆抗体可有效抑制子孢子对宿主细胞的入侵。这些结果说明该蛋白可能参与了虫体在宿主细胞内的生长发育、耐药性的产生以及子孢子入侵宿主细胞的过程。     

Long‐Term Tillage and Cropping System Effects on Chemical and Biochemical Characteristics of Soil Organic Matter in a Mediterranean Semiarid Environment

Several studies have reported how tillage and cropping systems affect quantity, quality, and distribution of soil organic matter (SOM) along the profile. However, the effect of soil management on the chemical structure of SOM and on its hydrophobic and hydrophilic components has been little investigated. In this work, the long‐term (19 years) effects of two cropping systems (wheat monoculture and wheat/faba bean rotation) and three tillage managements (conventional, reduced, and no tillage) on some chemical characteristics of SOM and their relationships with labile carbon (C) pools were evaluated. Soil samples were taken from the topsoil (0–15 cm) of a Chromic Haploxerert (central Sicily, Italy). After 19 years of different tillage and cropping systems management, total organic C significantly differed among treatments with the labile organic C pools showing the greater amount in no till and in wheat/faba bean plots. Hydrophobic and hydrophilic components of SOM, determined by diffuse reflectance infrared Fourier transform spectroscopy, were mainly affected by cropping system, whereas aromatic components of SOM by tillage. Soil organic matter components and characteristics showed significant correlations with the soil biochemical parameters, confirming the expected synergism between chemical and biochemical properties. This study demonstrated that (i) no tillage and crop rotation improve the chemical and biochemical properties of SOM of Vertisols under semiarid environment; and (ii) tillage management and cropping systems have affected, after 19 years, more the chemical and biochemical properties of SOM than its quantity. Copyright © 2014 John Wiley & Sons, Ltd.     

Study on Genetic Fingerprinting Technique of Microbial Communities

The 99% of all microorganisms in naturecannot be isolated by standard culture methods without conspicuous external characters to classify them morphologically. In addition, classification based on physiological or biochemical features are often fuzzy. Molecular techniques are used for uncultured microorganisms to explore the diversity of microbial communities and novel resources without the need of cultivation. DNA dynamics and cloning of PCR products obtained from environmental DNA is now routinely used to identify microbial populations in microbial ecology. For practical reasons the approach is not suited to study the complex dynamics of microbial communities in environment. Genetic fingerprinting techniques are better tools, which give complement for the traditional microbiological methods. The methods are rapid and relatively easy to perform, and they allow the simultaneous analysis of multiple samples, which makes it possible to compare the genetic diversity of microbial communities from different habitats, or to study the behaviour of individual communities over time.     

太岁的研究进展

太岁是一种多菌类复合体,其生物活性成分在抗癌、抗菌、细胞毒性及生理调控等方面具有重要功能.由于生物技术限制,目前对不同太岁样品的生物活性成分及菌群结构只做了初步分析,通过多种方式分离鉴定太岁菌种构成,对做好太岁的资源化利用具有重要的意义,现在距规模化开发利用太岁还有许多开创性的工作要做.通过综述太岁的微生物组成、药用价值、生物活性成分及其在微生态制剂方面的应用,对太岁应用前景进行展望,以期为太岁的进一步研究与利用提供借鉴法.     

园林植物土壤酶活性和微生物活性对盐碱胁迫的响应

以园林植物石楠为试材,通过混合2种中性盐(NaCl和Na2SO4)和2种碱性盐(NaHCO3和Na2CO3)分别模拟不同强度的盐、碱胁迫条件,对园林植物(石楠)进行3年(2015—2017年)的胁迫处理,以期探讨盐碱胁迫对园林植物土壤酶活性和微生物的影响。结果表明:1)土壤中有机酸含量随着盐碱浓度的增加呈逐渐降低趋势。随胁迫强度增加,盐、碱胁迫均使土壤中Na⁺浓度大幅度上升,K⁺浓度大幅度下降,但碱胁迫下Na⁺浓度上升幅度和K⁺浓度下降幅度均大于盐胁迫。2)随着盐碱浓度的增加,土壤中氨基酸类、碳水化合物类、羧酸类、聚合物类、胺类、酚酸类碳源呈逐渐降低趋势,总体而言,碳水化合物和羧酸类碳源是盐碱胁迫下园林植物土壤微生物的主要碳源,其次为氨基酸类、酚酸类和聚合物类,胺类碳源的利用率最小。3)土壤微生物物种丰富度指数(H)、均匀度指数(E)和碳源利用丰富度指数(S)、土壤微生物量碳、微生物呼吸和微生物代谢熵随盐碱浓度的增加而增加。4)土壤糖苷酶、几丁质酶、亮氨酸氨基肽酶、碱性磷酸酶、酚氧化酶δ和过氧化物酶δ随盐碱浓度的增加呈逐渐降低趋势。5)交互作用分析显示,盐胁迫、碱胁迫、盐胁迫×碱胁迫对有机酸、Na⁺、K⁺、Na⁺/K⁺、物种丰富度指数(H)、碳源利用丰富度指数(S)、土壤微生物量碳、土壤微生物呼吸、土壤微生物代谢熵、糖苷酶和碱性磷酸酶具有显著的影响(P<0.05),其中碱胁迫对土壤酶活性和微生物群落多样性的影响高于盐胁迫,并且盐胁迫×碱胁迫的影响高于单独盐胁迫或碱胁迫.     

鸡粪沼液复合微生物有机肥在温室樱桃番茄上的应用效果研究

为了探讨鸡粪沼液复合微生物有机肥在樱桃番茄上的田间应用效果,在滴灌施肥的条件下,设置化肥和液体复合微生物有机肥两个处理进行对比应用,结果表明施用液体复合微生物有机肥的处理其单果重可提高39.69%,含糖量可提高11.94%,茎粗可提高8.76%,Vc含量可提高5.72%,产量可提高30.95%,节约成本1.32%,可有效促进作物生长,提高果实着色均匀度,提高作物抗病性。同时,土壤速效氮提高6.87%,土壤速效磷46.23%,土壤速效钾提高16.86%,有机质含量提高12.64%,为作物提供了良好的生长环境。说明液体复合微生物有机肥对作物产量、品质以及土壤改良均有显著的提升作用,具有一定的应用前景。     

Avian pathogenic Escherichia coli MT78 invades chicken fibroblasts

Avian pathogenic Escherichia coli (APEC) are responsible for extraintestinal diseases, called colibacillosis, in avian species. The most severe manifestation of the disease is colisepticemia that usually starts at the respiratory tract and may result in bird death. However, it is not yet clear how APEC cross the respiratory epithelium and get into the bloodstream. In this work, we studied the interaction between 8 APEC strains (UEL31, UEL17, UEL13, UEL29, MT78, IMT5155, IMT2470, A2363) and a chicken non-phagocytic cell, the fibroblast CEC-32 cell line. We investigated the association profile, the invasion capability, the cytotoxicity effect and the induction of caspase-3/7 activation in an attempt to understand the way the pathogen gains access to the host bloodstream. Association to cells was determined after 1 h of infection, while cell invasion was determined after 4 and 24 h of infection. The cytotoxic effect of bacterial infection was measured by lactate dehydrogenase (LDH) release and the activation of the apoptotic program was verified by caspase-3/7 activation. Also, the presence of genes for adhesins, invasins and other related virulence-associated factors was verified by PCR. All bacterial strains showed similarity in relation to adhesion, LDH release and caspase-3/7 activation. However, one APEC strain, MT78, showed high invasion capability, comparable to the invasive Salmonella typhimurium strain SL1344. Since an APEC strain was capable of invading non-phagocytic cells in vitro, the same may be happening with the epithelial cells of the avian respiratory tract in vivo. CEC-32 monolayers can also provide a useful experimental model to study the molecular mechanisms used by APEC to invade non-phagocytic cells.     

一种水产养殖用微生物制剂的研究

利用枯草芽孢杆菌、干酪乳杆菌、沼泽红假单胞菌和啤酒酵母4种菌株配制了一种复合微生物制剂,试验结果表明,该制剂除含有丰富的营养成分外,在水产养殖中还能起到净水剂的作用,它能增加水中的溶解氧,显著降低氨氮、亚硝态氮、硫化物等不利于水产养殖的污染成分。此微生物制剂在水产养殖中将能发挥营养、保健及净化水质的功效。     

酵素菌肥施用量对日光温室土壤及芹菜产量品质的影响

通过田间试验研究了酵素菌肥不同施用量对日光温室土壤、芹菜产量及品质的影响。结果表明:供试土壤有机质、碱解氮、速效磷、速效钾、全盐含量随酵素菌肥施用量的增加而显著增加,pH则随施用量的增加而显著降低;在15～45t/hm~2施肥范围内,随酵素菌肥用量的增加,芹菜产量线性增加,VC含量略下降,可溶性糖含量显著增加,芹菜体内硝态氮含量显著升高,但仍属于A级绿色蔬菜。     

微生物叶面肥改善龙冈茌梨果实品质的研究

以10 a生龙冈茌梨为材料,研究了不同浓度的微生物叶面肥在其不同发育时期喷施处理对其外观品质和内在品质的影响.结果表明:微生物叶面肥能显著地提高龙冈茌梨的单果重,减小果皮厚度;但是低浓度处理对其果皮色泽和光洁度、果锈数量以及口感的改善效果不明显,只有在高浓度时才稍有改善.微生物叶面肥能显著提高龙冈茌梨果实的可溶性总糖含量以及可溶性固形物含量,能显著地降低龙冈茌梨果实的可滴定酸含量和石细胞含量,但是对果实硬度的改善并不明显.综合不同浓度微生物叶面肥对龙冈茌梨果实外观品质和内在品质的影响,认为较高浓度有助于提高龙冈茌梨的品质,以1∶600处理的效果最好.