水霉拮抗菌的筛选及其拮抗作用的初步研究

为了获得水产动物水霉病病原菌的拮抗微生物,以水霉为靶标,对其拮抗菌进行了筛选,并研究了拮抗作用最强菌株的拮抗活性.实验结果表明:从发生过水霉病害的水体中分离得到了130株细菌,能抑制水霉菌落生长的有三株:LD038、LD057和LD106,其中以LD038的拮抗作用最强,经梅里埃ATB微生物自动鉴定系统鉴定为黏质沙雷氏菌(Serratia marcescens).培养基对LD038的拮抗活性影响显著,LD038在PDA平板上表现出的拮抗活性最强,大小依次为PDA＞BHI＞GY＞CA＞Sabouraud.进一步对菌株LD038体外拮抗活性进行了研究.平板抑制实验表明,LD038能抑制水霉菌丝生长和孢子萌发.琼脂扩散法中,LD038菌落周围产生的抑菌圈直径为20mm,而无菌水对照组中水霉菌丝蔓过滤纸纸片并长至平板边缘;D38菌线间孢子的萌发与孢子距菌线的距离有关,距LD038菌线22mm范围内,孢子的萌发完全被抑制.无菌发酵上清液实验表明,与对照组相比,LD038无菌发酵上清液对孢子的萌发和菌丝生长都表现出显著的抑制作用.其中孢子较菌丝对上清液更为敏感,5倍稀释上清液下的孢子萌发率仅为10%,且萌发后的菌丝生长也同样受到抑制,整个实验过程中菌丝均未形成网状,对照组中萌发形成的菌丝在16h后呈网状.无菌上清原液和2倍稀释上清液中孢子均未萌发.对菌丝而言,仅无菌上清原液能彻底抑制菌丝生长,与对照组相比,2倍和5倍稀释上清液显著延缓了菌丝的生长.当对照组中的菌丝铺满整孔时,2倍和5倍稀释上清液中的菌丝长度分别为0和2.6mm.显微观察表明LD038的无菌发酵上清液导致水霉菌丝形态发生变化,较正常菌丝短而粗大,且出现了黑色原生质聚集.本研究为水霉病害的生物防治提供了一定的理论基础和依据.     

杀真菌素链霉菌AL-04的筛选与鉴定

对13株来源于青藏高原土壤的放线菌进行活化,再分别采用琼脂扩散法、生长速率法、孢子萌发抑制法和离体叶片法筛选出一株抗菌活性高且抗性稳定的菌株AL-04。抗菌活性结果表明,该菌株对4种常见的土传病害病原真菌:辣椒疫霉病菌(Phytophthora capsici)、黄瓜枯萎病菌(Fusarium oxysporum f.sp.cucumerinum)、西瓜枯萎病菌(Fusarium oxysporum f.sp.niveum)、番茄灰霉病菌(Botrytis cinerea)都有较强的抑制效果,抑菌率均在70.0%以上,其中对辣椒疫霉病菌(Phytophthora capsici)抑菌率高达93.0%。通过形态特征、生理生化特征分析和16S rDNA序列分析,将该菌株鉴定为杀真菌素链霉菌(Streptomyces fungicidicus)。     

芦荟内生菌内生哈茨木霉LH-7对植物病原菌的抗性

通过体外培养方法,研究了药用植物库拉索芦荟内生真菌内生哈茨木霉LH-7对9种常见植物病原菌的抑菌谱及其拮抗机理.结果表明： 内生哈茨木霉LH-7 对供试植物病原真菌具有显著的抑制作用(抑制率为62.4%～88.4%),且营养竞争和重寄生是其主要拮抗机制.该菌株的代谢产物中具有能够抑制病原菌生长及孢子萌发的活性物质,其作用可导致病原菌菌丝体生长的畸形、细胞壁破裂、产孢结构发育不健全、孢子形成及萌发率降低,从而有效抑制病原菌的生长和繁殖.
     

小花假泽兰茎叶中杀菌活性成分的研究

以小麦纹枯病原菌为指示菌种,采用活性跟踪法,从小花假泽兰(M ikan ia m icrantha H.B.K.)茎叶中分离出1种对黄瓜霜霉、黄瓜炭疽、小麦赤霉和玉米大斑4种病原菌孢子萌发具有较好抑制作用的物质,在供试浓度为0.50 m g.mL-1时,对黄瓜炭疽、小麦赤霉、玉米大斑3种病原菌孢子萌发抑制率均在70%以上.生长速率法测定其对番茄灰霉病原菌的EC50为0.039 m g.mL-1,组织法测定结果表明该化合物对番茄灰霉病有较好的防效,在供试浓度为2 000 m g.k-g 1时,治疗作用和保护作用防效分别达到了68.6%和48.7%,高于或相当于对照药剂速克灵的防效.经熔点测定、IR、M S1、HNM R1、3CNM R分析,鉴定该化合物为豆甾醇,其为首次从小花假泽兰茎叶中分离得到.     

天然植物提取物对茄子黄萎病菌的抑制活性

以茄子黄萎菌为供试菌种,研究了54种天然植物的乙醇提取液对茄黄萎菌的抑菌活性. 结果表明, 有15种植物提取物对茄子黄萎菌菌丝生长抑制率大于50%,其中细辛、黄连、厚朴、儿茶、苦参、甘草、蛇床子、桔梗、洋葱的提取物对供试菌菌丝生长抑制作用较强,抑制效果均在65%以上,蛇床子的抑菌率达86.84%.有16种植物提取物对病菌孢子萌发的抑制作用大于70%,其中7种植物提取物对孢子萌发的抑制率大于95%以上,细辛、黄连、厚朴基本上抑制了孢子的萌发.儿茶提取物对菌丝生长的抑制效果较差,但却显著抑制了孢子萌发,抑制率达100%.蛇床子提取物虽然抑菌效果较高,但其处理的孢子萌发率为11.3%.研究发现天然植物提取物中存在着抑制黄萎菌的活性物质,这为生物防治黄萎病和进一步研究蔬菜抗病增产理论提供了新的思路与途径.     

水霉拮抗菌的筛选与拮抗物特点初步分析

水霉病是淡水和咸淡水鱼类被水霉菌感染引起的常见疾病。该研究从多个养殖水体中分离得到6株水霉拮抗菌,SZK15、XL03、TG08、DJ12、HD05和DS08,以SZK15的拮抗作用最强。对SZK15研究发现,在PDA平板上,其对水霉菌丝抑制距离为11.5 mm,对水霉孢子抑制圈直径为29.7 mm,其10倍浓缩的无细胞发酵液的抑水霉菌圈直径达30.2 mm;在液体培养条件下,SZK15无细胞发酵原液能完全抑制水霉菌丝和孢子的萌发生长,其5倍稀释液中,水霉菌丝生长受抑制而长度稳定在2.0 mm,有30%的孢子萌发,且萌发后菌丝受抑制而未形成菌丝网,其10倍稀释液中水霉菌丝生长慢于对照组。SZK15分泌的拮抗活性物质对121℃高温敏感,对80℃、100℃高温、蛋白酶K和胰蛋白酶均部分敏感。本研究为水霉病害的生物防治提供了潜在新途径。     

富贵竹黑腐病拮抗菌H5的抑菌机制及相关特性研究

从湛江红树植物-红海榄筛选出1株对引起富贵竹病害黑曲霉病菌(Aspergillus niger)有良好抑制作用的菌株编号为H5,初步鉴定为地衣芽孢杆(Bacillus licheniformis).该菌株采用平板对峙、生长速率法及菅碟法对黑曲霉均表现良好的抑制作用,发酵原液对茵丝生长和孢子萌发的抑制率分别为91.9%和100%,抑菌带内菌丝扭曲畸形、表面产生大量囊状突起.其无菌滤液对酸碱、热稳定性较好,经55%硫酸铵沉淀后经121℃处理25 min仍能保持大部分活性,初步推测该物质为一种耐热的蛋白.     

臭氧水浸种对黄瓜种子萌发及幼苗生长的影响

臭氧水处理10、30和50 min后的黄瓜种子,种子萌发及幼苗生长均受到促进,种子的发芽率和活力指数、萌发种子的过氧化氢酶(CAT)和淀粉酶的活性以及呼吸速率、幼苗的壮苗指数和根系活力均有提高;处理70和90min的黄瓜种子萌发和生长则受到抑制,种子电解质外渗,过氧化氢酶和淀粉酶活性下降.     

石蒜对萝卜、黄瓜、番茄和油菜幼苗的化感效应

在室内用离体生测方法研究了石蒜水浸提物对萝卜、黄瓜、番茄和油菜的化感效应.结果表明, 石蒜水浸提物对4种植物种子的萌发和幼苗生长均具有较强的抑制作用.4种受体作物中,以番茄最为敏感,最低浓度0.0125 g·ml^-1处理下,番茄种子完全受到抑制不能萌发,油菜、萝卜与黄瓜种子的萌发抑制率分别为17.73％、14.97％和2.65％.相同浓度处理时,石蒜水浸提物对萝卜、黄瓜及油菜芽的生长抑制作用高于对根的抑制作用.采用去胚乳小麦生长法和高梁河沙法测定了石蒜甲醇浸提物对光合作用和非光合作用抑制活性.结果表明,石蒜甲醇浸提物对去胚乳小麦和高梁的生长有较强的抑制作用,对高梁芽和根生长的抑制作用高于对去胚乳小麦根和芽生长的抑制,说明石蒜对植物的化感效应主要体现在抑制非光合作用活性上,但也一定程度地抑制植物的光合作用.     

土壤丝状真菌中能引起稻瘟霉分生孢子形态畸变的活性菌种筛选的 …

本实验对安徽,湖南,江苏等十三个省的６０份土样进行了分离纯化,获得７９１株真菌。采用目前属国内领先的以稻瘟霉分生孢子生态变化为指标的筛选体系,初筛得到能使稻瘟霉孢子畸变和抑制孢子萌发的菌株１４０株,进而经过复筛得么能够使稻温霉孢子畸变的１９株,抑制稻瘟霉孢子萌发的１６株,其中７６－２,７６－６,１００－３,１００－７,１００－１０,５株菌的孢子畸变活性最好且遗传稳定。     

Antibacterial potentiality of Argemone mexicana solvent extracts against some pathogenic bacteria

The sensitivity of two Gram positive (Staphylococcus aureus and Bacillus subtilis) and two Gram negative (Escherichia coli and Pseudomonas aeruginosa) pathogenic multi-drug resistant bacteria was tested against the crude extracts (cold aqueous, hot aqueous, and methanol extracts) of leaves and seeds of Argemone mexicana L. (Papaveraceae) by agar well diffusion method. Though all the extracts were found effective, yet the methanol extract showed maximum inhibition against the test microorganisms followed by hot aqueous extract and cold aqueous extract.     

黄精种子营养成分的测定与分析

采用化学分析的方法,对黄精(Polygonatum sibiricum Red.)种子主要营养成分--粗淀粉、粗脂肪、粗蛋白的含量进行了测定,并采用气相色谱法、氨基酸自动分析仪分别对其脂肪酸和氨基酸的组分作了分析,以明确黄精种子的营养成分及其利用价值.结果显示:(1)黄精种子的营养成分以淀粉(73.70%)为主,脂肪含量为(10.74%),蛋白质含量为(8.21%);(2)脂肪酸中以不饱和脂肪酸(89.52%)为主,其中人体必需的多不饱和脂肪酸亚油酸占49.77%;(3)氨基酸种类齐全,其中人体必需氨基酸蛋氨酸、赖氨酸、异亮氨酸的含量比小麦、玉米等作物的含量高.结果说明,黄精种子具有较高的营养价值,在食品、医疗保健方面具有一定开发前景.     

苍耳等48种植物提取物的杀虫活性

采用浸虫法测定了48种植物的丙酮提取物对小菜蛾（Plutella xylostella L．）的杀虫活性,并研究了苍耳（Xanthium sibiricum Patrin．）不同溶剂提取物对小菜蛾和棉蚜（Aphis gossypii Glover）的触杀作用以及苍耳丙酮提取物不同溶剂萃取物对小菜蛾的触杀作用。结果表明,在供试的48种植物中,500g·L^-1黄花蒿（Artemisia annua L）、泽漆（Euphorbia helioscopia L．）、夏至草[Lagopsis supina（Steph．ex Willd．）Ik．-Gal．ex Knorr．]、苍耳及律草[Humulus scandens（Lour．）Merr．]的丙酮提取物对小菜蛾的杀虫活性较强。在苍耳的不同溶剂提取物中,苍耳丙酮提取物的杀虫活性最强,乙醇和乙酸乙酯提取物的杀虫活性次之。另外,50g·L^-1苍耳丙酮提取物的氯仿萃取物对小菜蛾的杀虫活性最强,24h和48h的校正死亡率分别为88．33％和91．67％。     

金银花和苦丁茶多糖提取物抗菌活性研究

从金银花和苦丁茶中提取水溶性多糖,利用圆形纸片法对7种常见的食品腐败菌进行抗菌试验。结果表明:金银花和苦丁茶多糖提取物都具有一定程度的抗细菌与抗真菌活性,抗细菌活性比抗真菌活性强;苦丁茶多糖提取物对金黄色葡萄球菌的最低抑菌浓度为5mg/ml。     

Mutagenicity screening of crude drugs with Bacillus subtilis rec-assay and Salmonella/microsome reversion assay

This paper describes the screening studies of 104 commercial crude drugs for mutagenicity by the rec-assay with Bacillus subtilis as well as the reversion assay with Ames strains TA98 and TA100 of Salmonella typhimurium. The rec-assays showed that 13 water extracts and 27 methanol extracts of the crude drugs were positive. The Ames assays with or without metabolic activation showed that 24 water extracts and 16 methanol extracts were mutagenic. In total, mutagenic activities were found in 45 samples among the 104 crude drugs tested.     

4种地衣提取物抗氧化和抗肿瘤活性研究

本研究初步评估了4种药用地衣(太白茶、金刷把、黑石耳、红石耳)不同溶剂提取物的抗氧化活性及其粗多糖的抗肿瘤活性。通过测定清除DPPH自由基、羟基自由基和还原能力,对4种地衣不同溶剂提取物进行体外抗氧化活性评价,结果表明,金刷把和黑石耳的甲醇提取相清除DPPH自由基能力高于其它溶剂提取相,其IC50值(半抑制浓度)分别为0.7847 mg/mL和0.5595 mg/mL;黑石耳甲醇提取相(IC50=0.5747 mg/mL)清除羟基自由基能力优于阳性对照物Vc(IC50=0.6126 mg/mL);黑石耳氯仿提取相、金刷把乙酸乙酯提取相和太白茶甲醇提取相清除羟基自由基能力与Vc相当;4种地衣甲醇提取相还原能力均较强,且与其浓度呈较好的量效关系。利用MTT法分析4种地衣多糖对HeLa、A375和Hep G2细胞体外生长增殖的抑制作用,结果显示黑石耳粗多糖对Hep G2细胞的抑制作用较为突出(IC50=0.2567 mg/mL),而金刷把抑制HeLa细胞的生长增殖作用最强,其IC50值为0.4332 mg/mL。     

Antioxidant activity of twenty five plants from Colombian biodiversity

The antioxidant activity of the crude n-hexane, dichloromethane, and methanol extracts from 25 species belonging to the Asteraceae, Euphorbiaceae, Rubiaceae, and Solanaceae families collected at natural reserves from the Eje Cafetero Ecorregión Colombia, were evaluated by using the spectrophotometric 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging method. The strongest antioxidant activities were showed by the methanol and dichloromethane extracts from the Euphorbiaceae, Alchornea coelophylla (IC50 41.14 mg/l) and Acalypha platyphilla (IC50 111.99 mg/l), respectively. These two species had stronger DPPH radical scavenging activities than hydroquinone (IC50 151.19 mg/l), the positive control. The potential use of Colombian flora for their antioxidant activities is discussed.     

Antifungal activity of phytochemicals from Eucalyptus hybrid leaves against some plant pathogenic and wood decay fungi

The in vitro fungitoxicity of the essential oil (EO), crude extracts, dried fractions and ursolic acid (UA) isolated from the abundantly available and underutilised leaves of Eucalyptus hybrid (Eucalyptus camaldulensis × Eucalyptus tereticornis) was assessed against three plant pathogenic fungi Aspergillus niger, Fusarium solani and Ganoderma lucidum causing pathogenesis in seeds, seedlings and trees, respectively, and Trametes versicolor and Pycnoporous sanguineus making wood to decay. The EO, methanol extract and UA demonstrated varied antifungal activity against all the fungi with minimum inhibitory concentrations ranging from 0.5 to 3.0%. The results of the study demonstrate a new chemical utilisation approach of the leaves of Eucalyptus hybrid towards the development of biofungicides in the management of above fungi of common occurrence in Indian forestry.     

Biological effects of active fraction isolated from Hydnocarpus pentandra (Bunch. –Ham.) Oken seeds against Helicoverpa armigera (Hub.) (Lepidoptera: Noctuidae)

Hexane, chloroform, ethyl acetate and methanol extracts of Hydnocarpus pentandra (Flacourtiaceae) seeds were tested for antifeedant, larvicidal, pupal mortality and adult deformations activities against Helicoverpa armigera. Crude extracts were screened at 0.5, 1.0, 1.5 and 2.0% concentrations. Bioassay-guided fractionation method was followed to isolate the active fraction from the crude extract. Active fraction was analysed by FT-IR, ¹H NMR, ¹³C NMR and GC-MS. Hexane extract presented the highest antifeedant (87.89%), pupal mortality (41.67%) and adult malformation activities at 2% concentration. Seven different fractions were isolated from hexane extract, among which fraction-2 showed the highest antifeedant (81.43%) activity and recorded the lowest LC₅₀ of 792.07 ppm. The fraction-2 contained two cyclopentenyl carboxylic acids, such as hydnocarpic acid (1) and chaulmoogric acid (2) in the ratio of 2:1. These compounds were major constituents in the active fraction of hexane extract of H. pentandra seeds. Fraction-2 can be used for agricultural pest management.     

A monoclonal antibody to (S)-abscisic acid: its characterisation and use in a radioimmunoassay for measuring abscisic acid in crude extracts of cereal and lupin leaves

A monoclonal antibody produced to abscisic acid (ABA) has been characterised and the development of a radioimmunoassay (RIA) for ABA using the antibody is described. The antibody had a high selectivity for the free acid of (S)-cis, trans-ABA. Using the antibody, ABA could be assayed reliably in the RIA over a range from 100 to 4000 pg (0.4 to 15 pmol) ABA per assay vial. As methanol and acetone affected ABA-antibody binding, water was used to extract ABA from leaves. Water was as effective as aqueous methanol and acetone in extracting the ABA present. Crude aqueous extracts of wheat, maize and lupin leaves could be analysed without serious interference from other immunoreactive material. This was shown by measuring the distribution of immunoreactivity in crude extracts separated by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), or by comparing the assay with physicochemical methods of analysis. Analysis of crude extracts by RIA and either, after TLC purification, by gas chromatography using an electron-capture detector or, after HPLC purification, by combined gas chromatography-mass spectrometry (GC-MS) gave very similar ABA concentrations in the initial leaf samples. However, RIA analysis of crude aqueous extracts of pea seeds resulted in considerable overestimation of the amount of ABA present. Determinations of ABA content by GC-MS and RIA were similar after pea seed extracts had been purified by HPLC. Although the RIA could not be used to analyse ABA in crude extracts of pea seeds, it is likely that crude extracts of leaves of several other species may be assayed successfully.Abbreviations ABA abscisic acid - DW dry weight - FW fresh weight - GC-ECD gas chromatography using an electron capture detector - GC-MS combined gas chromatographymass spectrometry - HPLC high-performance liquid chromatography - McAb monoclonal antibody - PVP soluble polyvinylpyrrolidone - RIA radioimmunoassay - TLC thin-layer chromatography