1株抗真菌放线菌菌株YIM31530T的多相分类研究

 从云南丽江玉龙雪山采集的土壤样品中分离到1株诺卡菌形稀有放线菌YIM31530T.该菌株具有很强的抗灰霉病、稻纹枯病活性.通过对其进行的包括形态、生理生化特性、细胞壁化学组分以及16SrDNA序列等方面的多相分类研究,确定其为高丽菌属的1个新种,将其命名为抗菌高丽菌(Kribbella antibiotica sp.nov.).该菌株在大多数培养基上生长良好,气生菌丝黄白色,基内菌丝浅黄色至黄白色.气生菌丝和基内菌丝多分枝,常断裂成不规则的杆状体,不游动.     

1株耐盐拟诺卡氏菌株的分离及初步鉴定

从青海高盐环境中分离到1株拟诺卡氏菌形放线菌YIM90039，该菌株具有典型的拟诺卡氏菌属的形态学特征．通过对其进行的形态学、生理生化特性、细胞壁化学组分以及16S rDNA序列等方面的分类学研究。菌株YIM90039初步鉴定为拟诺卡氏菌属的一个潜在新种．该菌株在大多数培养基上生长良好，气生菌丝大都呈黄白色，基内菌丝浅橙黄色至深橙黄色．气生菌丝上有长短不一的孢子链，基内菌丝则长，多分枝。常断裂成杆状。     

具抗肿瘤活性中度嗜盐菌YIM 80186的分离和系统发育分析

 从青海盐碱土壤样品中分离到1株中度嗜盐放线菌YIM 80186,该菌株的代谢产物具有很强的抗肿瘤和抗菌活性.菌株YIM 80186在添加20~100 g/L NaCl的培养基上生长良好,而在未添加NaCl的多数培养基上生长微弱.最高能耐受200 g/L NaCl.气生菌丝和基内菌丝丰富,白色至黄白色.气生菌丝产生直的或弯曲的孢子链,基内菌丝发育良好,断裂.生长pH范围6.0~10.0,最适pH 7.5~8.0.通过对其进行的包括形态、生理生化特性、细胞化学以及基于16S RNA基因序列的系统发育分析等方面的多相分类研究,该菌株具有拟诺卡氏属的典型特征,初步鉴定其为拟诺卡氏菌属的1个潜在新种.     

日本亮耳菌对植物病原真菌的拮抗试验

 首次报道了日本亮耳真菌对植物病原真菌的拮抗活性.结果表明:除对竹荪疫霉(Phytophthora sp.)的拮抗方式是菌丝缠绕作用外,该菌株的菌丝体及其发酵液滤液对灰葡萄孢(Botrytis cinerea)、尖镰孢菌(Fusarium oxysporum)、绿色木霉(Trichoderma viride)、烟草疫霉(Phytophthora nicotianae)和稻瘟病菌(Pyricularia oryzae)均有很强的拮抗活性.     

Alkalibacterium sp. SL3中α-半乳糖苷酶基因的克隆、表达和性质研究

以大布苏盐碱湖分离菌Alkalibacterium sp. SL3的DNA为模板,利用PCR扩增α-半乳糖苷酶基因(galSL3),构建重组质粒pET-22b-galSL3,转化至大肠杆菌BL21（DE3）诱导表达重组酶（rGalSL3）. 通过镍柱亲和层析分离纯化重组酶,并对纯化的重组酶进行性质研究. 研究表明,纯酶rGalSL3最适pH值为5.5,最适温度为55℃;该酶在pH值 5.0~10.0保持90%以上的剩余酶活,在50℃有非常好的热稳定性. 在0~1.5mol·L^-1 NaCl溶液中该酶的酶活基本不受影响,在3.0mol·L^-1 NaCl溶液中有很好的稳定性. Pb²⁺、Ca²⁺、Co²⁺、Li⁺、Na⁺、K⁺、Triton-100和β-mercaptoethanol等对重组酶的活性有明显的促进作用. 该酶水解对硝基苯基-α-D-吡喃葡萄糖苷的K_m、v_max和k_cat分别为(2.64±0.02)μmol·mL^-1、(454.55±0.59)μmol·(mg·min)^-1和(347.73±1.27)s^-1. 重组酶可水解蜜二糖和棉籽糖,不能水解瓜尔豆胶.     

Clostridium butyricum蔗糖磷酸化酶的酶学性质及其功能研究

本研究克隆表达丁酸梭菌Clostridium butyricum DSM10702菌株的蔗糖磷酸化酶基因,并探究重组酶的酶学性质和转糖苷功能。从丁酸梭菌C.butyricum DSM10702菌株中克隆蔗糖磷酸化酶基因cbsp,构建重组表达质粒pQE30-cbsp并在大肠杆菌Escherichia coli XL1-blue中诱导表达;将重组蛋白经镍柱纯化后,进行酶学性质和转糖苷功能研究。实验结果表明,以蔗糖为底物,重组酶Cbsp的最适温度为45℃、最适pH为7.0、V_max和K_m分别为(957±23.29) μmol·mg^-1·min^-1和(62.4±4.749) mmol·L^-1。当以葡萄糖-1-磷酸(G-1-P)为糖基供体时,重组酶Cbsp对于多数单糖、糖醇类、(-)-儿茶素、己基-β-D-吡喃葡萄糖苷和L-抗坏血酸表现出转糖苷活性。以上研究结果可丰富蔗糖磷酸化酶相关数据库,为其应用研究提供参考。     

利用Brevibacillus laterosporus YMF3.00003菌株研制生物杀菌剂

 Brevibacillus laterosporus YMF3.00003菌株具有广谱抗真菌活性,摇瓶培养发酵液对所测的植物病原菌Cylindrocarpon didynum,C.destructans,Magnaporthe grisea,Rhizoctonia solani的菌丝生长有较强的抑制作用,最强抑制率分别为81.5%,71.0%,60.7%和60.2%.确定了该菌株在20 L发酵罐中的发酵条件,绘制了生长曲线及得到了不同生长时期的抑菌率.其中,培养21 h时细胞数最多,达到5.1×109;25 h时发酵物对R.solani的抑菌率最高,达到90.5%.通过对几种吸附剂的筛选,确定用锯木屑作为吸附剂,将发酵终产物用锯木屑吸附,制成约2×10¹¹g^-1的菌剂.     

白黄链霉菌TD-1菌株挥发性有机物对灰霉孢菌的抑制作用

为探讨白黄链霉菌TD-1菌株挥发性有机物(VOCs)对灰霉孢菌的抑制作用,采用双皿对扣法测定VOCs对灰霉孢菌的抑菌率,通过扫描电镜观察VOCs对灰霉孢菌菌丝体的影响,利用GC/MS技术对VOCs组分进行分析。结果表明,作用5d后,6.0g TD-1菌株麦麸培养物产生的VOCs对灰霉孢菌的抑菌率为66.67%,灰霉孢菌的产孢量(3.9×10⁶个/cm²)较空白组(5.9×10⁷个/cm²)显著降低(p<0.01),且菌丝形态异常、表面粗糙,菌体细胞膜通透性增强。TD-1菌株VOCs由63种组分组成,其中2-甲基异冰片(13.52%)和1,4-二甲基金刚烷(14.08%)的相对含量较高。白黄链霉菌TD-1菌株产生的VOCs对灰霉孢菌良好的抑制作用为将其开发成生物熏蒸剂提供了理论依据。     

西藏砂生槐茎内链格孢属真菌生物学特性研究

为了解西藏豆科槐属植物砂生槐内生菌的生物学特性,本研究从砂生槐植株茎内分离得到链格孢属真菌,利用PDA培养基和KB培养基鉴定菌株菌落形态,利用载片培养法鉴定菌株孢子形态。结果显示:1菌株在PDA培养基上菌落舒展,呈白色毡帽状,等径生长,边缘呈波状;该菌株菌落到后期时,从菌落中心到边缘呈现三种不同颜色,即中心部位为白色,白色菌落边缘为黑色,最外层为暗绿色;气生菌丝茂密,孢子与基质均呈黑色。在KB培养基上菌落舒展,呈白色毡帽状,等径生长,边缘整齐;该菌株菌落到后期时,从菌落中心到边缘呈现三种不同颜色,即中心部位为白色,白色菌落边缘为暗黄色,最外层为暗绿色;气生菌丝茂密,孢子为黑色,基质呈黄色。2孢子呈倒棒状、球形、倒梨形和椭圆形等多种形态,具有纵、横或斜的真隔膜,呈砖格状分隔,喙多数为单细胞假喙,少数为短柱状假喙形态。3所分离出的菌株不具有性型,能形成简单的子座,分生孢子链从子座上伸出,并作合轴式延伸。4分离得到的菌株属于半知菌亚门丝孢纲丝孢目暗色孢科链格孢属真菌的一种。     

1株碱杆菌属菌株的分离和鉴定

 从新疆采集的盐土样品中分离到1株革兰氏阳性、严格好氧、产孢、嗜盐细菌菌株YIM 012,对其进行了系统分类学研究.菌株YIM 012为杆菌,生长最适温度为28~45℃,最适pH 7.0~8.0,并能在70~250g/L的盐质量浓度中生长良好.其DNA的G+C的摩尔分数为39%,16S rRNA基因序列分析结果显示,该菌株与碱杆菌属Alkalibacillus salilacus(DSM 16460^T)亲缘关系最近,相似性达到99.6%,但与同属其它2个有效种间相似性低于96%.杂交结果显示其与A. salilacus亲缘关系低于20%,并且在生理生化方面也有差异.综合分析,确定该菌株为碱杆菌属的1个新种,将其命名为嗜盐碱杆菌(Alkalibacillus halophilus).     

放线菌F401菌株杀虫活性研究

从江苏茅山地区竹林土壤中分离筛选到一株具有杀虫活性的放线菌F401号菌株。在扫描电镜下,该菌株的孢子丝呈紧密和松敞的小螺旋形,孢子椭圆形及圆柱形,表面光滑。气生菌丝灰褐色,菌落薄绒状,基内菌丝无色,不产生可溶性色素。摇瓶发酵菌丝抽提物稀释后对桑蚕(Bombyxmori)、大叶黄杨尺蠖(Abraxasmiranda)、褐刺蛾(Setorapostornata)及松褐天牛(Monochamusalternatus)等昆虫进行测试,均具有较强的杀虫活性。试验研究表明,此菌株是一种具有杀虫活性的链霉菌(Streptomicessp.)。     

Resistance of oat to ‘take-all’ causing fungus (Gaeumannomyces graminis var.tritici)

Evaluation with the pot assay at seedling stage in greenhouse showed that oat (Avena sativa) was highly resistant to take-all disease to which, however, wheat (Triticum aestivum) was extremely susceptible. The oat roots were shown to be inhibitory to the invasion and spread of take-all causing fungus G. graminis var. tritici by the following criteria: (ⅰ) less infection sites were observed (about 1/7 of those in wheat); (ⅱ) the ectotrophic growth of G. graminis var. tritici on oat roots was much slower than that on those of wheat, and the runner hyphae appeared as kidney- or fork-shaped hyphopodia on the surface of oat roots which could not be discerned on that of wheat roots; (ⅲ) the period from inoculation to penetration into the epidermis of oat roots was about 2.9 times as long as that of wheat; (ⅳ) the infection hyphae were hindered substantially when it was about to penetrate into the epidermis of oat roots with the mycelium deformed; and (ⅴ) the cortical layer of oat roots was revealed to be unsuitable for the G. graminis var. tritici infestation as some lysed hyphae were found therein, and the spread of hyphae from the first layer of cortex to the pericycle needed 108 h, about 1.8 times as long as it did on wheat roots.     

Molecular regulation of development and differentiation inStreptomyces

Development and differentiation is an important and leading research field in modem biology. Streptomyces has a complicated life cycle of morphological differentiation including the spore germination, aerial mycelium and spore formation. Each developmental stage has a distinguished morphological feature which greatly facilitates the identification of developmental mutants, the complementary cloning and the spatial and temporal expression of the genes involved in differentiation. This characteristic of Streptomyces  is comparatively superior to other prokaryotic bacteria such as Escherichia coli, Bacillus subtilis and Myxococcus xanthus. Moreover, Streptomyces  also possesses a complicated physiological differentiation in which it produces a wide variety of secondary metabolites (more than half of the 12 000 or so known antibiotics), including many important antibiotics used in medicine, agriculture and industry. Studies on the molecular mechanism of antibiotic biosynthesis will be helpful in improving the antibiotic producer and developing some new medicines. In comparison with eukaryotic microorganism such as Asperillus nidulans,the structure of genetic material in Streptomyces  is simple,and it is linear and conkaryotes such as Streptomyces  cerevisiae. The large number of genes are the molecular basis of Streptomyces  differentiation,suggesting that the regulation mechanism of gene expression in differentiation and development may be complex^[1]     

不同处理对赤苍藤种子萌发和幼苗生长的影响

为研究不同因素对药食两用植物赤苍藤(Erythropalum scandens Bl.)种子萌发和幼苗生长的影响,本研究以当年采收的赤苍藤种子为材料,探讨浸种时间(0 d、1 d、2 d、3 d、4 d、5 d)、基质含水量(70%、50%、30%)、播种深度(0 cm、1 cm、3 cm、5 cm)、光照条件(全黑、自然光)、基质(沙土、肥土、黄土、混合土)、赤霉素浸种处理及破壳处理(全破、少破、不破)下赤苍藤种子的发芽率及幼苗特性,为其规模化发展提供理论参考。结果表明,浸种时间、基质含水量、播种深度、赤霉素处理均对发芽率有极显著影响(P<0.01),基质和破壳处理对发芽率有显著影响(P<0.05),浸种5 d、50%和30%的基质含水量、播种深度1 cm、混合土基质、50 mg·L^-1赤霉素浸种12 h以及全破处理的发芽率在各对比的处理中均最高。浸种时间和基质含水量对幼苗叶长和枝高均有显著影响(P<0.05),其他处理对幼苗生长无显著影响。综上可知,赤苍藤种子育苗时,可先浸种5 d或用50 mg·L^-1赤霉素浸种12 h...     

Foraging strategies in the greater horseshoe bat （Rhinolophus f errumequinum ）on Lepidoptera in summer

Abstract The diet of the bat Rhinolophus ferrumequinum was studied at the Zhi＇an Village of Ji＇an City in China, from June to August 2004. The bats were trained in a laboratory （volume： 9×4×4 m^3）. Foraging strategies of the bat were observed at night and prey remains were collected and identified. The results showed that the diet consisted mainly of Lepidoptera in summer, including 11 families, more than 30 species of moths, such as Noctuidae （36.6% by number）,Sphingidae （24.1%）, Geometridae （13.4%） and Limacodidae （9.5%）. The length of culled wings ranged from 10--40 mm （97.7%）. Pearson correlation analysis showed that the bat R.ferrumequinum foraged their prey selectively, but not opportunistically. From field studies, two ways were observed in which the bats retrieved their prey including aerial hawking during peak active period of the insects and flycatching during the insects＇ non-peak activity period. The bats never gleaned prey from the ground, though they appeared to be well able to detect fluttering moths on the ground.     

Inhibition of Alkaline Phosphatase from Pearl Oyster Pinctada fucata by o-Phthalaldehyde： Involvement of Lysine and Histidine Residues at the Active Site

Alkaline phosphatase from Pinctada fucata was inactivated by o-phthalaldehyde （OPA）. The inactivation followed pseudo first-order kinetics with a second rate constant of 0.167 （mmol/L）^-1·min^-1 at pH 7.5 and 25℃. A Tsou＇s plot analysis showed that inactivation occurred upon formation of one isoindole group. The OPA-modified enzyme lost the ability to bind with the specific affinity column and the presence of substrates or competitive inhibitors protected the enzyme from inactivation. The results revealed that the OPA-reaction site was at the enzyme substrate binding site. Prior modification of the enzyme by lysine or histidine specific reagent abolished formation of the isoindole derivatives, suggesting that lysine and histidine residues were involved in the OPA-induced inactivation. Taken together, OPA inactivated the alkaline phosphatase from Pinctada fucata by cross-linking lysine and histidine residues at the active site and formed an isoindole group at the substrate binding site of the enzyme.     

阴地蕨、银粉背蕨总黄酮和总酚含量及其抗氧化能力分析

测定了阴地蕨(Botrychium ternatum (Thunb) Sw.)和银粉背蕨(Aleuritopteris argentea(Gmel.) Fee)乙醇提取物的总黄酮和总酚含量,及其对1,1-二苯基-2-三硝基苯肼(DPPH)自由基的清除能力、2,2'-联氮基双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基的清除能力和Fe的还原力.结果表明:1)阴地蕨和银粉背蕨乙醇提取物样品中总黄酮质量分数在4. 2～41. 2 mg·g~(-1)之间,总酚质量分数在3. 9～16. 4 mg·g~(-1)之间,两者地上部分总黄酮和总酚含量均高于地下部分的,阴地蕨与银粉背蕨的总黄酮和总酚含量存在一定差异; 2)阴地蕨和银粉背蕨的乙醇提取物对DPPH、ABTS自由基具有一定的清除能力及还原力,且提取物抗氧化活性与总黄酮和总酚含量存在浓度依赖.     

榕树须醇提取物对脑血栓形成和凝血功能的影响

【目的】探讨榕树须不同提取物的抗脑血栓作用,并评价其对凝血功能的影响。【方法】采用小鼠脑偏瘫、电刺激致大鼠血栓模型观察提取物对小鼠脑血栓的保护率、大鼠血栓形成时间的影响;采用肾上腺素加冰浴致小鼠血瘀模型观察其对凝血酶时间及纤维蛋白原时间的影响。【结果】榕树须95%、80%醇提物的不同提取部位能提高小鼠脑偏瘫恢复率(20%~60%);乙酸乙酯及正丁醇部位对电刺激引起的脑血栓有较好保护作用(P0.01,P0.001);乙酸乙酯部位及正丁醇部位均能延长寒凝血瘀小鼠的凝血酶时间(P0.05,P0.001);正丁醇部位能延长寒凝血瘀小鼠的纤维蛋白原时间(P0.05)。【结论】榕树须80%醇提物的正丁醇部位具有较好的抗脑血栓及改善血瘀小鼠凝血功能的作用。