一种野生多孔菌的分离、鉴定、培养条件及抗氧化活性

多孔菌是一类子实体呈孔状且质地为革质至木质的大型担子菌,其中一部分具有较高的药用价值.对一株野生多孔菌子实体进行分离纯化获得纯培养BJ菌株,并对其分类、最适培养条件和液体发酵产物抗氧化活性进行分析.采用形态学和ITS分类学鉴定菌株的分类学地位;通过测定菌株在不同碳源、氮源等培养基中的生长状况,研究菌丝最适培养条件;使用2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)法测定菌株发酵液总抗氧化活性;使用总超氧化物歧化酶(T-SOD)法、1,1-二苯基-2-苦基肼基自由基(DPPH)法测定菌株发酵液菌体的总超氧化物歧化酶活力和自由基清除能力.结果显示:经鉴定BJ菌株为石榴嗜蓝孢孔菌(Fomitiporia punicata).菌丝体最适培养碳源为葡萄糖、麦芽糖和淀粉,最适氮源为酵母浸粉,最适C/N比为10/1,最适温度为28℃,最适pH为7.0.发酵液总抗氧化活性为0.517 mmol/L(维生素E),菌体的总超氧化物歧化酶活力为770.37 U/g,DPPH自由基清除力的IC_(50)为2.14 mg/mL.本研究从野外获取了一株高抗氧化活性的药用多孔菌资源,可为野生药用真菌的开发利用提供理论依据.     

米曲霉产氨基酰化酶条件及部分酶学性质研究

通过单因子和多因子摇瓶正交优化试验,确定了米曲霉液态发酵产氨基酰化酶的最佳发酵条件.优化发酵培养基组成(ρ/gL-1):葡萄糖40,蔗糖10,可溶性淀粉20,蛋白胨2.5,马铃薯液1000mL,pH自然.培养基装量50mL/250mL三角瓶,接种量4%.培养温度30℃,转速100r/min,发酵时间42h.每50mL培养物的总酶活由优化前的2627u提高到7338u,是优化前的2.79倍.研究了米曲霉氨基酰化酶的部分酶学性质.该酶催化反应的最适pH为7.0,最适温度为40℃,低浓度的Co2 (5×10-4mol/L)对酶活激活作用显著.图5表2参8     

纤维素酶产生菌HS-F9的筛选鉴定和产酶条件优化

为了选育高产纤维素酶菌株,通过刚果红鉴别培养基以及滤纸条崩解实验测定,从牛粪堆肥中筛选到一株产纤维素酶的真菌HS-F9,根据菌株形态特性和18SrRNA基因序列分析,初步鉴定该菌为绿色木霉(Trichoderma viride).利用液体发酵培养产生纤维素酶,研究了碳源、氮源、培养时间、培养温度、培养基起始pH、接种量对菌株HS-F9产酶的影响.结果表明:产EG、CBH和FPA的最适碳源均为CMC-Na;EG和CBH在以蛋白胨为唯一氮源时酶活最高,FPase则在以黄豆粉为唯一氮源时酶活最高;产生EG、CBH和FPA的最适温度分别为30℃、30℃、33℃;最适起始pH为3.0、3.0和4.0;EG和FPase的最适接种量为2%,CBH最适接种量达到了8%;培养时间均以5～6d为宜.在最适条件下培养,该菌株EG、CBH和FPase的酶活分别达到了5275.3U/mL、8502.1U/mL和3619.1U/mL,是未优化前的1.42、1.35和1.32倍.图6表2参23     

枫杨种子和幼苗对地下水位及埋藏深度的响应

通过室内试验研究不同地下水位(0、-2、-4、-6、-8和-10cm)和不同埋藏深度(0、1、2、3、4、5和6cm)条件下枫杨(Pterocarya stenoptera)种子的萌发特性以及幼苗的建成状况.结果表明,种子萌发的最适水位为0～-4 cm;幼苗定植与生长的最适水位为-6cm.随埋藏深度增加,枫杨出苗率下降,出苗时间推迟,幼苗的生物量根冠比增大.试验第11周,各埋藏深度条件下幼苗的地上株高无显著差异,埋藏深度对幼苗的生长也无显著影响.枫杨种子萌发及萌发后幼苗生长的最适埋藏深度为1～3cm.     

黑胸散白蚁纤维素酶的体外酶学特性

采用DNS法研究了我国广泛分布的一种低等木食性白蚁——黑胸散白蚁纤维素酶的体外酶活特性以了解其纤维素降解机制.结果表明,内切β-1,4-葡聚糖酶(EG)、纤维二糖水解酶(CBH)和β-葡萄糖苷酶(BG)这3种酶的最佳反应时间均为15 min,最佳底物浓度为1%,最适反应pH为5.6,最适反应温度为35℃.在最适反应条件下,EG、CBH和BG的活性分别达到71.3(±13.9)U/mg、5.8(±0.8)U/mg和4.1(±0.7)U/mg.EG在体外的热稳定性较差,在50℃及更高温度酶活很低或完全失活,但该酶对pH稳定性较好,在pH 3.2～8.0范围内酶活力变化不大.Native-PAGE电泳检测到该白蚁体内至少有8种不同的EG活性条带,肠道不同部位纤维素酶活性条带种类不同.这些研究表明,木食性白蚁降解纤维素是一个复杂的过程,需要多种纤维素酶的共同作用.     

一株野生桑黄的分离鉴定与生物学特性

桑黄是我国传统名贵的药用真菌,在分类学上是锈革孔菌科(Hymenochaetaceae)几种药用真菌的总称.从北京延庆区四海镇黑汉岭采集到一株桑黄子实体,经分离纯化获得纯培养(编号SS).进一步开展其分类鉴定、最适培养条件和液体发酵产物活性研究.根据ITS鉴定,确定SS菌株属于锈革孔菌科针层孔菌属(Phellinus),与苹果木层孔菌(Phellinus tuberculosus)的相似性为95%;结合子实体、菌丝特征和寄主植物类型,确定SS菌株为苹果木层孔菌.SS菌株菌丝体最适培养条件研究表明,其最适碳源为葡萄糖,最适氮源为黄豆粉,最适C/N比为20/1,最适生长因子为维生素C,最适温度为28℃,最适pH为7.0.以马铃薯葡萄糖液体(PD)培养基、10%接种量、28℃、150 r/min培养7 d,发酵液中多糖含量为176.71 mg/L,黄酮含量为0.11 mg/L,抗氧化活性为7.82 mol/L(FeSO_4),未检测到多酚的存在.本研究从野外获取新的桑黄药用资源,其生物学特性结果可为野生药用真菌的人工驯化和开发利用提供理论依据.     

一株白腐菌产生的漆酶对RB亮蓝的脱色作用

W 1是一株能在液体条件下产漆酶的白腐菌 ,纯化的漆酶对RB亮蓝有很好的脱色作用 .漆酶的最适脱色温度为 4 5℃ ,最适脱色pH值为 6 .0 ,脱色pH范围在 4～ 7之间 .当溶液中漆酶活力为 2 .0× 10 3 U/L时 ,在最适脱色条件下、16h内 ,RB亮蓝 (30 0mg/L)的脱色率可以达到 90 % .经酶作用后 ,RB亮蓝在 4 30～ 70 0nm范围内的特征颜色吸收峰基本消失 .实验证明 ,在相同的条件下 ,漆酶粗酶对RB亮蓝有更好的脱色效果 .图 7表 1参 6     

芒苞草（芒苞草科）种子的无菌萌发研究

选取野外采回的芒苞草种子为材料,研究无菌水和不同状态Ms培养基以及不同激素种类和浓度对芒苞草种子萌发的影响,并初步探讨了芒苞草幼苗的生长情况.结果表明:经4℃的低温春化处理2 mo后,播种在无菌水中的芒苞草种子萌发率可达95%,比同样条件下MS液体(25.1%)和固体(10.4%)培养基中的萌发率高;在有关激素影响的实验中,无菌水中6-BA和GA,对种子萌发没有促进作用,液体MS培养基中的1.0 mg/L GA,明显促进种子萌发,而固体MS培养基中0.5 mg/L 6-BA对种子萌发的促进作用较明显,1.0 mg/L 6.BA有利于从生芽的诱导.将无菌水和液体MS培养基中已发芽的芒苞草种子转移至固体MS培养基中培养可长成小植株.     

除草剂阿特拉津(Atrazine)溶液中弹琴蛙蝌蚪(Rana adenopleura)的行为模式

研究了不同条件下阿特拉津溶液中 (ρ1～ 4/mgL-1:5 ,10 ,15 ,2 0 )弹琴蛙 (Ranaadenopleura)蝌蚪的行为 ,运用方差分析 (ANOVA)检验了蝌蚪在不同浓度的阿特拉津、时间段、pH值、溶氧量等条件下的行为差异性 .根据实验结果 ,确立了正常及异常行为模式 .结果表明 :在水温 (15 .0± 0 .5 )℃的条件下 ,弹琴蛙蝌蚪异常行为的出现主要与阿特拉津的浓度有关 ,同时与实验时间的长短也有一定的关系 ,而与 pH值、溶氧量关系不大 .实验组与对照组相比 ,蝌蚪出现异常行为的频次差异极显著 (P <0 .0 1) ;其中在实验组中 ,蝌蚪在 5mg/L的阿特拉津溶液中与 10mg/L、15mg/L、2 0mg/L中出现异常行为的频次差异极显著 (P <0 .0 1) ,而在 10mg/L、15mg/L、2 0mg/L阿特拉津溶液中出现异常行为的频次差异不显著 (P >0 .0 5 ) ,相关性分析表明 ,蝌蚪异常行为的出现与阿特拉津的浓度没有剂量反应关系 (r2 =0 .4 94 ) ;在对照组中 ,0 .5mL/L丙酮对照和空白对照中蝌蚪出现异常行为的频次差异不显著 (P >0 .0 5 ) .图 3表 2参 17     

一株嗜酸性产纤维素酶真菌的特性及产酶条件优化

从四川海螺沟原始森林腐土中分离到一株嗜酸性产纤维素酶的真菌X-13,其主要特点是产纤维素酶的最适pH及其纤维素酶最适反应pH均为2.0.在PDA培养基上培养时菌落呈浅黄色至肉桂色,反面呈黄色至棕褐色,产黄色色素;菌丝体透明有隔膜,分生孢子呈球形或近球形.根据菌株的形态特征以及ITS序列同源性和系统发育分析结果,鉴定该菌株为土曲霉(Aspergillus terreus Thom).该菌最佳产酶培养时间为8～10 d;最适产酶温度为30℃,纤维素酶最适反应温度为50℃;最佳碳源、氮源分别为纤维素粉和硫酸铵.通过响应面法对菌株产纤维素酶条件进行优化,使菌株X-13纤维素酶活从1.39 IU/mL提高到2.94 IU/mL,提高了111.5%.     

模拟酸雨对外来入侵植物三叶鬼针草生长及化感作用的影响

外来入侵植物三叶鬼针草（Bidens pilosa L.）对我国华南地区生态系统造成严重危害,同时华南地区也是强酸雨分布区域。采用不同pH值（2.5,4.0,5.6）的模拟酸雨对三叶鬼针草种子和幼苗进行处理,测定其对种子萌发和幼苗生长的影响;室外模拟酸雨喷淋三叶鬼针草,生物测定三叶鬼针草叶片水浸提液对受体植物的化感作用的变化。结果表明：pH为4.0和5.6的酸雨处理后提高了三叶鬼针草种子的萌发率;pH为4.0的酸雨促进幼苗生长,增加幼苗苗高和生物量;而pH为2.5的酸雨抑制三叶鬼针草种子萌发率和幼苗生长。同时,pH 4.0和2.5酸雨增强了三叶鬼针草叶片水浸提液对受体植物的化感作用。研究表明：三叶鬼针草具有较强的耐酸能力,酸雨胁迫可能通过促进外来入侵植物三叶鬼针草的生长和提高化感潜力而有利于其进一步入侵。     

拮抗菌B96-Ⅱ对芦笋枯萎菌的抑菌作用

为明确拮抗菌B96-Ⅱ对芦笋枯萎菌的抑制效果并探明其抑菌特性及方式,对经B96-11处理后芦笋枯萎菌的生长速率、液体电解质、电导率及菌丝重量等进行了测定研究.结果表明,在10~7～10~8 CFU/mL时,B96-Ⅱ对芦笋枯萎菌均有显著的抑制效果.连续3 wk室内观察发现,B96-Ⅱ对芦笋枯萎菌丝有持续的抑制作用,与对照比较抑制率为94.07%～88.98%.田问防治效果为93.40%.而农药多菌灵处理的防治效果仅为65.12%.拮抗菌B96-Ⅱ对芦笋枯萎菌的抑制方式主要为:(1)对孢子形成的抑制作用;(2)对孢子的溶解作用;(3)对菌丝生长的抑制作用;(4)对菌丝的致畸作用;(5)对菌体细胞的破损作用.B96-11处理后36 h,溶液总溶解性同体和电导率增高,而菌丝重量下降.图4表3参14     

Seed germination in response to chemicals: effect of nitrogen and pH in the media

Seed germination generally presents a peak in the next growing season after a fire. Among other factors associated with fire are the increase of soil nitrogen and changes in the pH of the soil. In this study, we addressed the question, whether or not the germination response of eight species is linked with the increase in pH and nitrogenous compounds in the germination media? We assessed the separate and combined effects of nitrogenous compounds and pH on the percentage and rate of germination of seeds of Medicago arabica (L.) Hudson, Epilobium hirsutum L., Foeniculum vulgare Miller, Daucus carota L., Thapsia villosa L., Cynosurus cristatus L., Dactylis glomerata L. and Rumex crispus L. All these species are well represented in the Mediterranean ecosystems of the central-west Spain. Water and CaCl2 were used as controls. Nitrogenous compounds increased percent germination (level) and rate in three of the species studied. High pH negatively affected the germination rate of seeds from most species, but had no effect on the per cent germination of any of the species. The higher concentration of the nutritious solutions affected negatively the germination level and rate. The different germination responses of seeds of the studied species could not be exclusively attributed to pH values in the media, whereas the amount and form of Nitrogen in the media has a greater effect on it. These differences in germination are species dependent.     

Removal of arsenic in tailings by soil flushing and the remediation process monitoring

In order to investigate the optimum conditions for the application of soil flushing of arsenic, a batch test was carried out using EDTA at various concentration and pH levels. Based on the optimum condition derived from the batch test, a column test was conducted to examine the feasibility of the soil-flushing technology under field-equivalent conditions. In this column test, a low flushing solution flow rate showed a significantly higher As removal efficiency (71.6%) than a high flow rate (56.3%). TCLP (toxicity characteristic leaching procedure) and a seed germination test were carried out to monitor the toxicity both during and after the treatment. The finally treated tailings were shown to be significantly remediated, having a reduced toxicity by both the TCLP and seed germination tests.     

Removal of arsenic in tailings by soil flushing and the remediation process monitoring

In order to investigate the optimum conditions for the application of soil flushing of arsenic, a batch test was carried out using EDTA at various concentration and pH levels. Based on the optimum condition derived from the batch test, a column test was conducted to examine the feasibility of the soil-flushing technology under field-equivalent conditions. In this column test, a low flushing solution flow rate showed a significantly higher As removal efficiency (71.6%) than a high flow rate (56.3%). TCLP (toxicity characteristic leaching procedure) and a seed germination test were carried out to monitor the toxicity both during and after the treatment. The finally treated tailings were shown to be significantly remediated, having a reduced toxicity by both the TCLP and seed germination tests.     

Response of peroxidase and catalase to acid rain stress during seed germination of rice, wheat, and rape

Seed germination of plants with various acid-resistance display different responses to acid rain. To understand the reason why such differences occur, the effects of simulated acid rain (pH 2.5–5.0) on the activities of peroxidase (POD) and catalase (CAT) during seed germination of rice (O. sativa), wheat (T. aestivum), and rape (B. chinensis var. oleifera) were investigated. Results indicated that the maximum change in activities of CAT and POD by acid rain treatment with different acidity and time in relation to the referent treatment without acid rain, was in the order: rice (28.8%, 31.7%) < wheat (34.7%, 48.3%) < rape (79.3%, 50.0%). The pH level for which the treatment with acid rain did not cause significant difference (p < 0.05) was in the order: rice (3.5).wheat (4.0).rape (5.0). Moreover, the change in activity of POD was higher than that of CAT, which showed that POD was more sensitive to acid rain stress than CAT. The difference in the ability of POD and CAT in removing free radicals was one reason why the germination indexes of these three species behaved differently.     

硅促进水稻种子萌发及缓解幼苗砷毒性的效应研究

通过As~(Ⅲ)胁迫下水稻种子的发芽试验和幼苗毒性试验,研究了外源硅对水稻种子发芽率、幼苗生长的影响及其缓解幼苗砷毒性的效应。外源硅的2种处理方式为种子萌发时添加外源硅(Si1)和采用硅处理液浸种(Si2)。结果表明,发芽时介质中As浓度达到10 mg·L~(-1)时显著抑制水稻种子萌发(P0.05),发芽率仅为80%,但是Si1和Si2处理下发芽率则提高到97%和100%,这说明外源硅可促进砷胁迫下水稻种子萌发;砷浓度≥5 mg·L~(-1)时,Si1和Si2处理均可提高水稻的相对幼苗高度和根耐性指数,提高幅度分别为6.00%~16.8%和57.9%~77.0%、7.10%~23.5%和54.2%~61.2%,并且降低了水稻幼苗砷含量,降低幅度分别为17.8%~21.4%和31.0%~49.1%。这说明外源硅处理可促进砷胁迫下水稻幼苗的生长;不同砷浓度处理与水稻芽长、根长及幼苗干重之间存在"S"型的剂量-效应关系,且外源硅显著提高了相应的EC50,缓解了砷对水稻幼苗生长的毒性。综上所述,砷胁迫下水稻种子萌发时添加外源硅或采用硅处理液浸种均可促进水稻种子萌发和幼苗生长,并降低了幼苗砷累积和缓解砷对水稻幼苗的毒性。     

<Emphasis Type="Italic">Bacillus algicola</Emphasis> decolourises more than 95% of some textile azo dyes

Textile effluents in natural waters pose environmental health problems if not treated to safe limits. Various bacterial species have the potential to degrade dyes. Here we studied the ability of Bacillus algicola to decolourise red, blue and yellow azo dyes. B. algicola was isolated from soil samples taken from a sanitary landfill site. Isolation and screening were performed using mineral salt medium. Dye-decolourising isolates were assessed in their capacity to decolourise dyes. Experiments were conducted at pH 6, 7 and 8, and 25, 35 and 45 °C. Phytotoxicity of the dyes and biodegradation products was assessed by seed germination tests. Results show that B. algicola gave the highest decolourisation at pH 8.0 and 25 °C in the presence of yeast extract as media supplement. B. algicola degraded the red and blue azo dyes by over 95%. The phytotoxicity results indicated that biodegradation products of the red and blue azo dyes were not toxic. Biodegradation products of the yellow dye were, however, toxic and considerably hindered germination. From these results, we infer that B. algicola has good potential for degrading and decolourising the red and blue test azo dyes.     

Impact of distillery effluent on germination behaviour of Brassica napus L

The study has been focused on effect of untreated distillery effluent (Devans Breweries Ltd., Jammu) on germination of gobi sarson (Brassica napus. L. var. Punjabi Special). Six treatments (E0.... E100) each having three replicates were made. E0 was taken as control in which tap water was used for irrigation of the plants. For E20, E40, E60, E80 and E100, different concentrations i.e. 20, 40, 60, 80 and 100% of effluent were used for irrigation, respectively. The 100% sample of distillery effluent analyzed for various physicochemical parameters showed acidic nature (pH 4.0) and higher values of COD (2496 mg l(-1)), TDS (799.7 mg l(-1)) and chlorides (1408 mg l(-1)). The parameters e.g. percent germination, germination index, speed of germination, and peak value were highest in treatment receiving 20% effluent concentration which also showed minimum values for percent inhibition, germination period, and delay index.     

一株中度嗜盐淀粉酶产生菌的分离鉴定及酶活测定

为开发极端环境工业用酶,从新疆盐碱土微生物中分离得到一株中度嗜盐高产淀粉酶活性菌株H3,其能耐受30%盐浓度和pH 11的极端环境.通过形态特征、生理生化实验及16S rRNA基因序列分析,确定H3属于Gracilibacillus属.该菌株能在盐浓度为0~30%的培养基上生长,最适生长盐浓度为5%~10%,最适pH值为8.5.在最适生长盐浓度、pH条件下,其淀粉酶活性可达到4 830个活力单位,可用于高盐高碱环境下淀粉的水解.