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1.
采用甲醇、乙酸乙酯和水分别按液土比3∶1、6∶1和9∶1对三七连作土壤进行浸提,研究其浸提液对三七根腐病菌生长和种群数量的影响。结果表明: 平板培养72 h后,甲醇、乙酸乙酯和水浸提液对尖镰孢菌和腐皮镰孢菌的菌丝生长均表现为化感促进,其中,甲醇和乙酸乙酯浸提液对尖镰孢菌的化感效应指数为14.0%~19.8%和16.2%~20.2%,高于水浸提液的8.9%~14.2%,且不同浸提比例之间差异不显著;而甲醇浸提液对链格孢菌菌丝生长表现为化感抑制,且抑制效应在浸提比例为3∶1时最强,达到-33.2%~-38.5%,乙酸乙酯和水浸提液对链格孢菌菌丝生长无显著影响。土壤培养4周后,甲醇、乙酸乙酯和水浸提液均能增加土壤中尖镰孢菌的数量,其中,水浸提液的增加效应最强,达到每克干土3.49×106~9.56×106拷贝数,高于甲醇(每克干土1.68×104~6.73×104拷贝数)和乙酸乙酯浸提液(每克干土1.77×104~3.72×104拷贝数),且这种增加效应随浸提比例的增加逐渐减弱;水浸提液和低浸提比例的甲醇提取液均能增加土壤中腐皮镰孢菌的数量,而重茬土壤浸提液对链格孢菌的数量影响不显著。因此,三七连作土壤浸提液对根腐病菌如尖镰孢菌和腐皮镰孢菌均表现出明显的化感促进效应,这可能是再植三七易发生根腐病等土传病害的原因之一。  相似文献   

2.
无机盐、激素与真菌联合诱导土沉香抗逆能力的研究   总被引:1,自引:0,他引:1  
为了探究无机盐、激素与真菌联合诱导土沉香(Aquilaria sinensis)抗逆性及其与结香前期木芯变色长度之间的相关关系,以10年生土沉香为材料,采用均匀试验设计,开展了3种无机盐、3种激素及3种真菌组合试验。结果表明:(1)土沉香树体POD和SOD活性和MDA含量呈先升高后降低趋势,CAT活性在诱导后1天最高,随后呈下降趋势;(2)无机盐与真菌处理下土沉香的抗逆能力大于激素与真菌处理,其中无机盐与真菌组合试验的处理4(1.0%CaCl2+0.5%FeSO4+2.0%NaCl+黑绿木霉∶腐皮镰孢∶龙眼焦腐(1∶1∶1))和激素与真菌组合试验的处理2(0.01%茉莉酸甲酯+0.1%乙烯利+0.2%水杨酸钠+黑绿木霉∶腐皮镰孢∶龙眼焦腐(1∶1∶1))分别为同类处理最高;(3)土沉香抗逆能力高低与木芯变色长度存在显著正相关性;(4)理论上,土沉香抗逆能力最强诱导组合分别为0.93%CaCl2+0.53%FeSO4+2.5%NaCl+黑绿木霉∶腐皮镰孢∶龙眼焦腐(1∶1∶1)和0.005%茉莉酸甲酯+0.006%乙烯利+0.2%水杨酸钠+黑绿木霉∶腐皮镰孢∶龙眼焦腐(1∶1∶1)。  相似文献   

3.
The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae. The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [FSP-FAP-cThEn(GC)] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [SP(HVChi26)-FSP-FAP-cThEn(GC)]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae. The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici, and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.  相似文献   

4.
采用体外α-葡萄糖苷酶抑制模型对一株球毛壳菌H6的发酵液和菌丝体两种乙酸乙酯提取物进行活性评价,结果表明,发酵液乙酸乙酯提取物对α-葡萄糖苷酶具有较强的抑制活性,其IC50值为(510.76±23.46)μg/mL。采用硅胶、Sephadex LH-20、半制备高效液相等色谱技术从其发酵液乙酸乙酯提取物中分离得到12个化合物。通过各种光谱分析,依次鉴定为chaetoviridins A-B(1-2),chaetoglobosins A-D(3-6),chaetoglobosin J(7),chaetoglobosin Q(8),prochaetogobosinsⅠ-Ⅲ(9-11),vibratilicin(12),其中化合物12为首次从毛壳属中分离得到。对化合物进行α-葡萄糖苷酶抑制活性测定发现,化合物12显示弱的抑制活性,其IC50为(1 182.75±19.14)μg/mL。  相似文献   

5.
毛筒壳科真菌次级代谢产物生物活性的评价   总被引:1,自引:0,他引:1  
毛筒壳科Tubeufiaceae真菌具有产新结构、新活性次级代谢产物的潜力,目前对该科真菌次级代谢产物的研究较少。为了寻找具有生物活性的新化合物,有必要对毛筒壳科真菌次级代谢产物及其活性进行系统深入的研究。本文采用平板对峙法、生长速率法和MTT法,分别测定已分离得到的19株该科真菌活体菌株抑菌活性、发酵物抑菌活性以及发酵物粗提物对不同人体肿瘤细胞株增殖的抑制作用。通过平板对峙法,试验共筛选获得13株活性菌株,其中,红棕毛筒腔菌菌株Tubeufia rubra PF02-2对7种植物病原真菌有明显的抑菌效果,抑制率均高于60%且抑菌谱广。采用生长速率法,发现红棕毛筒腔菌菌株PF02-2经液体发酵后,发酵液对其中4种植物病原真菌仍有一定的抑制作用,且菌丝体部分的乙酸乙酯提取物对马铃薯早疫病病菌Alternaria solani(ZYB)的抑制效果最好。通过MTT法,发现发酵物粗提物对3种肿瘤细胞均具有不同程度的细胞毒活性,其中在300μg/mL时,剑叶莎毛筒腔菌菌株Tubeufia machaerinae ML03-2发酵液部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa和人前列腺癌细胞株PC-3的抑制率(%)分别达到了98.92±0.15和97.86±0.18,在400μg/mL时,对人肝癌细胞株HEPG2的抑制率(%)达到了98.88±0.04;在500μg/mL时,明孢新旋卷孢菌菌株Neohelicosporium hyalosporum ML05-1菌丝体部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa细胞的抑制率(%)为98.32±0.02,在600μg/mL时,对人肝癌细胞株HEPG2的抑制率(%)达到了97.62±0.20,在300μg/mL时,对人前列腺癌细胞株PC-3的抑制率(%)达到了98.91±0.02。该研究结果为开发利用毛筒壳科真菌提供了科学依据。  相似文献   

6.
A method is described for measuring (-)-threo-chlorocitric acid in human plasma. Plasma is acidified to pH 1 to minimize lactonization and a13C analogue of (-)-threo-chlorocitric acid is added as internal standard. The acidified plasma is then extracted with ethyl acetate containing 10% methanol. The ethyl acetate—methanol extract is back-extracted with acetate buffer (pH 5). This extract, following adjustment to pH 1, is reextracted with ethyl acetate. The residue after removal of the ethyl acetate is treated with ethereal diazomethane. The wet residue is reconstituted in ethyl acetate and a portion of this solution is analyzed by gas chromatography—chemical ionization mass spectrometry. The mass spectrometer is set to monitorm/z269 [MH+ of trimethylated (-)-threo-chlorocitric acid] andm/z270 [MH+ of trimethylated (-)-threo-[13C]chlorocitric acid] in the gas chromatographic effluent. Them/z/269 tom/z270 ion ratio in a sample containing an unknown amount of (-)-threo-chlorocitric acid is converted to an amount of compound using a calibration curve. The calibration curve is generated by analyzing control plasma spiked with various known amounts of (-)-threo-chlorocitric acid and a fixed amount of (-)-threo-[13C]chlorocitric acid. The limit of quantitation is 0.1–0.6 μg ml−1, depending on the characteristics of the calibration curve generated with each set of samples. The precision (relative standard deviation) at a concentration of 2 μg ml−1 is 3.3%.  相似文献   

7.
采用石油醚、乙酸乙酯、乙醇浸提朱红栓菌 Trametes cinnabarina 子实体干粉,得到不同极性提取物;采用清除DPPH 自由基、羟自由基、超氧阴离子自由基能力,测定提取物的体外抗氧化活性;MTT法检测提取物对人肝癌细胞株HepG2细胞增殖的抑制作用。结果表明,朱红栓菌石油醚、乙酸乙酯、乙醇提取物均具有一定的抗氧化、抗肿瘤活性;各提取物在浓度为4-5mg/mL时,对DPPH自由基、羟自由基和超氧阴离子自由基清除能力大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物对3种自由基的最高清除率分别为60.23%、74.49%、63.84%。各提取物对人肝癌细胞株HepG2细胞增殖抑制作用大小依次为乙酸乙酯提取物>乙醇提取物>石油醚提取物;乙酸乙酯提取物的抑制率最高达55.93%。采用硅胶和凝胶等柱色谱方法结合核磁、波谱和质谱等技术对乙酸乙酯提取物的化学组分进行分析,共分离纯化出11种化合物,分别鉴定为:麦角甾醇(1),邻苯二甲酸二丁酯(2),对羟基苯甲酸甲酯(3),麦角甾-7,22,二烯-3-酮(4),1-[(12E,16E)-12,16-二十碳二烯酰基]-2-[(E,E)-7,11-十八碳二烯酰基]-3-硬脂酰基甘油(5),cinnabarin(6),过氧麦角甾醇(7),尿嘧啶(8),甘露醇(9),腺嘌呤核苷(10),豆甾-7,22-二烯-3β,5α,6β-三醇(11)。除化合物6外均为首次从朱红栓菌子实体中分离得到。研究结果为开发利用朱红栓菌子实体提供了科学依据。  相似文献   

8.
海洋真菌能够产生大量活性独特的次级代谢产物。为了探明海绵共生真菌产黄青霉LS16发酵液中抗副溶血弧菌Vibrio parahemolyticus的活性物质,本实验对副溶血弧菌Vibrio parahemolyticus的抑菌活性进行跟踪,采用VLC(vacuum liquid chromatography)、Sephadex LH-20柱层析、薄层层析和高效液相色谱等技术,从海绵共生真菌LS16乙酸乙酯发酵液中分离纯化得到5个化合物。进一步实验证明,化合物2具有抗副溶血弧菌Vibrio parahemolyticus活性。根据该化合物的波谱数据(1H NMR、13C NMR)对其化学结构进行鉴定,确定其分子式为C15H15NO3,为生物碱类化合物。  相似文献   

9.
李伟滔  贺闽  陈学伟 《植物学报》1983,54(5):547-549
由真菌Rhizoctonia solani引起的纹枯病严重危害玉米(Zea mays)和水稻(Oryza sativa)等作物的安全生产。R. solani的宿主范围广且抗源少, 加之相关的抗性机制研究有限, 导致纹枯病的危害长期得不到有效控制。近期, 中国科学家通过对318份玉米自交系进行全基因组关联分析, 筛选到1个与纹枯病抗性相关的、编码F-box结构域蛋白的候选基因ZmFBL41 (GRMZM2G109140)。ZmFBL41蛋白是SCF (SKP1-Cullin-F-box) E3泛素连接酶复合体的一员, 能介导复合体对肉桂醇脱氢酶ZmCAD的降解, 从而降低木质素的积累, 使玉米易感纹枯病。玉米抗病自交系Chang7-2中, 蛋白ZmFBL41 Chang7-2因2个关键氨基酸的变异, 不能结合并降解底物ZmCAD, 使木质素含量增加, 从而提高玉米对纹枯病的抗性。该研究率先揭示了SCF复合体可通过降解肉桂醇脱氢酶来调控植物免疫反应的新型分子机制, 为提高玉米及其它作物对纹枯病的抗性提供了重要理论依据和基因资源。  相似文献   

10.
11.
李伟滔  贺闽  陈学伟 《植物学报》2019,54(5):547-549
由真菌Rhizoctonia solani引起的纹枯病严重危害玉米(Zea mays)和水稻(Oryza sativa)等作物的安全生产。R. solani的宿主范围广且抗源少, 加之相关的抗性机制研究有限, 导致纹枯病的危害长期得不到有效控制。近期, 中国科学家通过对318份玉米自交系进行全基因组关联分析, 筛选到1个与纹枯病抗性相关的、编码F-box结构域蛋白的候选基因ZmFBL41 (GRMZM2G109140)。ZmFBL41蛋白是SCF (SKP1-Cullin-F-box) E3泛素连接酶复合体的一员, 能介导复合体对肉桂醇脱氢酶ZmCAD的降解, 从而降低木质素的积累, 使玉米易感纹枯病。玉米抗病自交系Chang7-2中, 蛋白ZmFBL41 Chang7-2因2个关键氨基酸的变异, 不能结合并降解底物ZmCAD, 使木质素含量增加, 从而提高玉米对纹枯病的抗性。该研究率先揭示了SCF复合体可通过降解肉桂醇脱氢酶来调控植物免疫反应的新型分子机制, 为提高玉米及其它作物对纹枯病的抗性提供了重要理论依据和基因资源。  相似文献   

12.
青霉属真菌Penicillium sp. CPCC 400786的抗病毒活性成分   总被引:1,自引:0,他引:1  
采用抗艾滋病毒抑制剂筛选模型对一株青霉属真菌Penicillium sp. CPCC 400786发酵产物的乙酸乙酯提取物进行活性评价,结果显示,其对艾滋病毒有较强的抑制活性。采用正相硅胶柱、Sephadex LH-20凝胶柱和半制备HPLC等色谱技术对乙酸乙酯提取物进行分离纯化,从中分离得到8个化合物。通过波谱数据分析,分别鉴定为:oxalicine A(1)、oxalicine B(2)、cis-4,6-dihydroxymellein(3)、亚油酸(4)、十八烯酸(5)、肉豆蔻酸(6)、尿嘧啶(7)、胸腺嘧啶(8)。化合物1和2为杂萜类化合物。对化合物1-6进行了抗艾滋病毒(HIV-1)和抗甲型流感病毒(H1N1)的活性评价。结果显示,化合物1具有良好的抗H1N1活性,其IC50值为38.5μmol/L,比阳性对照药利巴韦林稍弱(IC50=20.5μmol/L);化合物1和2具有抗HIV-1的活性,其IC50值分别为22.4、67.8μmol/L;其他化合物未显示抗病毒活性。本研究为从青霉属中发现更多抗病毒活性杂萜分子提供了依据。  相似文献   

13.
Genetic engineering has improved the product yield of a variety of compounds by overexpressing, inactivating, or introducing new genes in microbial systems. The production of flavor-enhancing ester compounds is an emerging area of heterologous gene expression for desired product yield in Escherichia coli. Isoamyl acetate, butyl acetate, ethyl acetate, and butyl butyrate are reported here to be produced by expressing Saccharomyces cerevisiae genes ATF1 or ATF2 and the strawberry gene SAAT in E. coli when the appropriate substrates are provided. Increasing the concentration of alcohol added to the reaction generally resulted in increased ester production. ATF1 expression was found to produce more isoamyl acetate and butyl acetate than ATF2 expression or SAAT expression in the strains and culture conditions examined. Additionally, SAAT expression resulted in greater isoamyl acetate and butyl acetate production than ATF2 expression. Butyl butyrate is produced by cell-free extracts of E. coli harboring SAAT but not ATF1 or ATF2.  相似文献   

14.
Traditional Chinese medicine (TCM) belongs to the most elaborate and extensive systems of plant-based healing. The herb Northern Ban Lan (Isatis tinctoria) is famous for its antiviral and anti-inflammatory activity. Although numerous components isolated from I. tinctoria have been characterized so far, their modes of action have remained unclear. Here, we show that extracts from I. tinctoria exert anti-microtubular activity. Using time-lapse microscopy in living tobacco BY-2 (Nicotiana tabacum L. cv Bright Yellow 2) cells expressing green fluorescent protein-tubulin, we use activity-guided fractionation to screen out the biologically active compounds of I. tinctoria. Among 54 fractions obtained from either leaves or roots of I. tinctoria by methanol (MeOH/H2O 8:2), or ethyl acetate extraction, one specific methanolic root fraction was selected, because it efficiently and rapidly eliminated microtubules. By combination of further purification with ultra-high-performance liquid chromatography and high-resolution tandem mass spectrometry most of the bioactivity could be assigned to the glucosinolate compound glucobrassicin. Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells. In the light of these findings, the antiviral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread.  相似文献   

15.
采用形态学及ITS-rDNA序列分析法,对具有抑制乙酰胆碱酯酶活性的蛇足石杉内生菌株JR14进行鉴定,确定为无柄盘菌Pezicula sp.。开展了乙酰胆碱酯酶抑制活性化合物的分离,从Pezicula sp. JR14乙酸乙酯提取物中分离到4个化合物,利用核磁共振及质谱技术将其鉴定为behenic acid (1)、himeic acid B (2)、secalonic acid A (3)和palmitic acid (4)。采用改进的Ellman比色法对分离的化合物进行活性检测,结果表明,himeic acid B (2)具有较强的乙酰胆碱酯酶抑制活性,其IC50为205μg/mL(0.64mmol/L)。  相似文献   

16.
The proteolytic extract obtained from the latex of Funastrum clausum (Jacq.) Schlechter (Asclepiadaceae), a South American climbing plant, was assayed as a novel catalyst for peptide synthesis and compared with commercial papain under the same conditions. After immobilization on polyamide, the synthesis of the bitter peptide precursor Z-Ala-Phe-OMe was performed and different conditions were tried. Acetonitrile and ethyl acetate with low water content were tested as organic solvents. Equilibrium- and kinetically-controlled synthesis were tried by using either Z-Ala-OH or Z-Ala-OMe as acyl donors, respectively. The best conditions for the synthesis of the desired product varied according to the catalyst used. For papain, thermodynamic control in acetonitrile (aw  0.12) in the presence of triethylamine (TEA) or boric acid–borate buffer (40 mM), and equilibrium- and kinetic-controlled synthesis in ethyl acetate (aw  0.75) proved to be the best conditions. The thermodynamic control in either acetonitrile with aw  0.12 (40 mM TEA or Na2CO3) or ethyl acetate (aw  0.75) were the best conditions found for funastrain. In all cases, the formation of oligopeptides up to three Phe was observed. The proteolytic extract of F. clausum latex showed more selectivity than papain towards the conversion to Z-Ala-Phe-OMe leading to less proportion of oligopeptides.  相似文献   

17.
Pseudomonas strains isolated from the rhizosphere of chickpea (Cicer arietinum L.) and green gram (Vigna radiata L.) were screened for the production of chitinases and cellulases. Five Pseudomonas strains were found to produce appreciable amounts of both enzymes in culture-free supernatants and showed growth inhibition of the two fungi Pythium aphanidermatum (Oomycete) and Rhizoctonia solani (Basidiomycete) in plates on potato dextrose agar medium. The fungal growth inhibition was not correlated with cell wall-degrading enzyme activity, which suggested that other antifungal compounds produced by these rhizobacteria were also involved in antagonism. Coinoculation of the Pseudomonas strains with the Mesorhizobium sp. Cicer strain Ca181 resulted in a significant increase in nodule biomass when grown under sterilized chillum jar conditions. The results suggest that hydrolytic enzymes produced by Pseudomonas sp. contribute to suppression of plant diseases by inhibiting growth of phytopathogenic fungi and also promote nodulation of legumes by rhizobia.  相似文献   

18.
Lipases from Candida rugosa, Mucor javanicus and Rhizopus oryzae were respectively adsorbed on Amberlite XAD-7 followed by incubation in 2-propanol and then lyophilization. The activities of the immobilized enzymes were 1.6–3.4 times higher than those of the immobilized enzymes without incubation in the organic solvent before lyophilization for esterification of lauric acid (0.1 M) and 1-propanol (0.1 M) in isooctane at 37 °C. The immobilized C. rugosa lipase (Sigma) without the incubation did not show any activity but displayed considerable activity (19.8 μmol h−1 mg−1) after the incubation before lyophilization. Besides 2-propanol, acetone, 1-propanol and ethyl acetate were also found to be good solvents for treating M. javanicus lipase immobilized on Amberlite XAD-7 and acetone was the best among them. When incubated in isooctane at 25 °C for 120 h, the immobilized M. javanicus lipase prepared by incubation in acetone for 1 h before lyophilization retained 70% of its initial activity while the immobilized enzyme without the solvent treatment kept only 50% of its initial activity.  相似文献   

19.
Microbial transformation of ferulic acid to acetovanillone was studied using growing cells of Rhizopus oryzae. Ferulic acid was added to the growing medium (0.5 g L-1) and incubated for 12 days. The progress of formation of metabolites was monitored by GC and GC-MS after extraction with ethyl acetate. The major metabolite was acetovanillone with minor metabolites formed, such as dihydroferulic acid, coniferyl alcohol and dihydroconiferyl alcohol. Traces of metabolites (≤1-3%), such as vanillin, vanillyl alcohol, vanillic acid and phenyl ethyl alcohol, were also produced. Formation of 4-vinyl guaiacol increased from day 1 (12.4%), reaching a maximum on day 4 (31.7%), and reducing to a minimum on day 12 (3.1%). The formation of acetovanillone increased only from day 2 onward, and reached a maximum (49.2%) on day 12. The optimum concentration of ferulic acid to be added into the medium was found to be only 0.5 g L-1, as any increase in concentration (0.75 and 1.0 g L-1) precipitated the precursor, resulting in no further degradation.  相似文献   

20.
Objective: The cell lines secreting specific monoclonal antibodies (McAbs) were prepared by using Fusarium solani, one of the pathogenic fungi causing root rot of Fritillaria thunbergii, and the colloidal gold immunochromatographic test strip based on McAbs was developed to provide scientific basis for detecting root rot of F. thunbergii. Methods: Hybridoma technology was used to obtain cell lines that could secrete specific McAbs against F. solani using the whole protein extract of F. solani as the antigen. The specificity, titer, sensitivity and binding protein of McAbs were detected by indirect ELISA and Western blot. Colloidal gold particles were prepared by trisodium citrate reduction method and McAbs were labeled to prepare colloidal gold immunochromatographic strip. Results: Three cell lines secreting specific McAbs against F. solani were obtained, which were named as FsA3, FsG6 and FsD4. The detection sensitivity of FsA3 was 24.41 ng / mL, and that of both FsG6 and FsD4 was 12.21 ng / mL. FsA3, FsG6 and FsD4 had strong reactions to F. solani, and had no cross-reaction to Alternaria tenuissima, A. alternata, Botrytis cinerea, F. equiseti, F. incarnatum, F. oxysporum, Phoma sp., and Phomopsis oblonga. The colloidal gold immunochromatographic strip based on FsG6 showed only a quality control line when detecting the tissue culture seedlings of F. thunbergii. When 100 ng F. solani antigen or the samples of F. thunbergii infested with root rot disease were detected, there were visible quality control lines and test lines. Conclusion: The specificity and sensitivity of the McAbs and test strip are sufficient to detect F. solani isolated from diseased strains of F. thunbergii, which provides the technical support for the rapid detection of root rot of F. thunbergii in the field.  相似文献   

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