47 resultados para FUNGUS GNATS


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齐墩果酸(OA)是一个分布广泛、含量丰富的天然三萜化合物,常以皂苷元的形式广泛存在于植物中,具有多种重要生物活性。但是OA许多活性较弱,且生物利用度低,限制了其在临床上的应用。一是OA水溶性差;二是抗癌活性仍与临床应用的抗癌药物相差比较大。 真菌在微生物转化中具有种类多、培养条件比较简单等特点,为了寻找到具有转化OA能力的菌株,采取一步发酵的方法,在18株实验室保藏真菌菌株中筛选到5株目的菌株,TLC分析显示有转化效果。 随后采用二步发酵的方法作为复筛,验证5株菌株转化能力,波谱分析结果表明5株菌株对OA确实有转化作用。 选择5株菌种中代号1F-2 2菌株作为放大实验菌株,分离转化产物,得到OA衍生物108(相对分子量414m/z)和1010(相对分子量340 m/z),分离出的产物用于活性检测。寻找到产物108的RP-HPLC分离条件,质谱得出二者相对分子质量。 为验证OA转化产物抗肿瘤活性,首次研究了OA对卵巢癌细胞株IGROV1和人乳腺癌细胞株MDA-MB-231作用,通过细胞增殖抑制实验、用MTT法检测细胞活性,结果表明齐墩果酸可降低卵巢癌细胞株IGROV1和乳腺癌细胞MDA-MB-231细胞增殖能力并呈剂量依赖性,对肿瘤细胞株的半数有效抑制浓度化IC50 分别为36.58μg/mL和38.8μg/mL (P<0.01)。OA能抑制肿瘤细胞活性,并且OA对卵巢癌细胞株IGROV1抑制活性高于乳腺癌细胞MDA-MB-231。 在此基础上,转化产物108和1010对卵巢癌细胞株IGROV1和人乳腺癌细胞株MDA-MB-231的抑制作用也进行研究,MTT实验结果表明,转化产物对两株癌细胞也有抑制活性(P<0.01)。 总之,本文工作为进一步开展齐墩果酸类化合物结构改造和抗肿瘤活性的研究奠定了基础。 Oleanolic acid (OA) is a triterpenoid widely distributed in the nature which possesses various important bioactivities. OA also serves as aglycon of many natural saponins. However, the relatively weak activities and poor bioavailability hinder its clinical use. Firstly, poor water-solubility results in worse bioavailability. Secondly, compared with clinical antitumor drug, the antitumor effect of OA has a great difference, it is worse. Many fungi have ability to transform nature products into a variety of derivatives, and transformation conditions of fungi are simple. Attempt to obtain fungi strains able to biotransform OA, we carried out the following experiments: To investigate the biotransformation 0f OA by strains supplied firstly, we used one-step fermentation method to screen the aimed strains from 18 fungus strains stored in our laboratory. On the basis of the initial screening experiments, we found 5 aimed strains. The TLC results showed that the 5 fungi strains could transform OA into other components derivatives. Then we used two-step fermentation method as secondly screening. We repeated the five strains to do the experiments, analytical data of the results proved the transformation indeed. In the followed experiments work, we chose 1F-2 2 strain as large-scale transformation fungus from the aimed fungi. We got two biotransformation products of OA by 1F-2 2, and named those derivatives 108 and 1010. We found RP-HPLC separation conditions of product 108. The two products were characterized by ESI-MS. To verify the anti-tumor activity of biotransformation products of OA, we studied the inhibition effect of oleanolic acid on the ovarian carcinomas IGROV1 and breast cancer cell line MDA-MB-231 firstly. With an assay based on a tetrazolium dye (MTT), the effects of various concentrations of oleanolic acid on ovarian carcinomas IGROV1 and breast cancer cell line MDA-MB-231 were studied. MTT method was used to measure the tumor cells viability. Compared with the control group, oleanolic acid can significantly inhibit the viability of the ovarian carcinoma cells IGROV1 and MDA-MB-231 breast cancer cell line (P<0.01), IC50 values were 36.58μg/mL or 38.8μg/mL. Oleanolic acid can inhibit the malignant tumor cells viability, and inhibitory activity of OA to ovarian carcinomas IGROV1 was higher than to breast cancer cell line MDA-MB-231. On this basis, we studied the anti-tumor activity of the two derivatives of OA [called 108 (414 m/z) and 1010(340 m/z)]. It came to the conclusion that the two derivatives also showed potent inhibitory effect on the growth of these tumor cells(P<0.01). Therefore, the results of studies will benefit the further investigating on the relationships of structures and antitumor activities of OA.

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本文筛选出一株能利用木糖产乙醇的丝状真菌Z7,对其利用木糖和半纤维素水解产物产乙醇的发酵条件进行了研究,并对Z7 利用玉米芯产木聚糖酶的条件进行了优化。全文分为三部分: 第一部分:目标微生物筛选、纯化及系统发育分析。以木糖为唯一碳源,采用梯度稀释和平板化线法从高温、中温酒曲中分离到16 株能利用木糖良好生长的丝状真菌;通过发酵试验复筛,获得一株能产乙醇的丝状真菌Z7;综合形态学和ITS 序列分析,初步鉴定为Aspergillus flavus。 第二部分:Z7 的乙醇发酵条件研究。以木糖为碳源,通过单因素试验确定最佳氮源和发酵温度;通过正交试验及SPSS 软件分析得到了不同N、P、K 成分对乙醇、残糖和菌体干重的影响。获得最佳的发酵条件为:(g/L)木糖50,尿素1, NH4NO3 1, K2HPO4 2 , KCl 0.5 , MgSO4.7H2O 0.5 , NaNO3 1 , pH 自然,培养温度33 ℃。以玉米芯半纤维素稀酸水解液为底物进行乙醇发酵,根据稀酸水解的单糖释放量和乙醇产量,确定115 ℃,1 h 为最佳玉米芯预处理条件;结合最佳发酵条件,添加1 g/L 的吐温20 能获得最大的乙醇浓度8.31 g/L。因此,Aspergillus flavus Z7 能利用半纤维素水解产物产乙醇,其中木糖的利用率80%以上。 第三部分:Z7 利用玉米芯产木聚糖酶条件优化。Aspergillus flavus Z7 在具有产乙醇能力的同时还具有产木聚糖酶的能力。本文通过单因素和正交试验得到最佳产酶培养基组分为:(g/L)玉米芯20,尿素2, 酵母膏2.5, K2HPO4 5,NaNO31, MgSO4.7H2O 1。单因素试验表明,用纱布代替塑料布密封摇瓶封口能显著提高产酶量;Z7 在碱性条件下具有更强的产酶性能。在最优条件下发酵,能产生最大木聚糖酶活122.23IU/mL。通过薄层分析,验证了Z7 产生的木聚糖酶具有水解木聚糖生成木糖及木寡糖的能力。 A strain of filamentous fungus which can produce ethanol by using the xylose was isolated in this research. The ethanol fermention conditions from xylose and dilute-acid hydrolyzate of the corn core were studied. The conditions of xylanase production by Z7 were also optimized. The paper involved three parts. Part1: Isolation, purification and phylogenetic analysis of the microbe. By using xylose as the single carbon source and the pla te streaking method, several filamentous fungi were isolated from the wine starter; through the fermentation test, a filamentous fungus Z7 which can produce ethanol was further recognized; furthermore, according to the morphologic observation and ITS seque nces analysis, Z7 was identified as Aspergillus flavus at the first step. Part2: Research on the condition of ethanol fermentation by Z7. By single factor experiment, the optional nitrogen resource and temperature of the fermentation were fixed; meanwhile, through the orthogonal array tests and the analysis of statistic software SPSS, the optional component of the culture medium and the fermentation condition were organized as follows: (g/L) xylose 50, urea 1, NH4NO3 1, K2HPO4 2, KCl 0.5 , MgSO4.7H2O 0.5, NaNO31, pH nature, temperature 33℃. Based on these optimal parameters, the fermentation of dilute-acid hydrolyzate of the corn core was carried on by Z7. According to the quantities of released sugar monomers and content of the ethanol, 115℃ in 1h is the best pretreatment condition; the maximal ethanol content can be obtained when 1g/L Tween 20 was added to. Therefore, the filamentous fungus Aspergillus flavus can use the hydrolysate of hemicellulose to produce ethanol, and the rate of xylose utilization was over 80%. Part3: Optimization of Z7’s xylanase producing condition from corn core. Aspergillus flavus Z7, which can utilize xylose or the hydrolysate of hemicellulose to produce ethanol, also had the ability of xylanase production. The optional component of the culture medium were fixed by the single factor experiment and the orthogonal array tests, and they were organized as follows: (g/L) corn core 20, Urea 2, Yeast extract 2.5, K2HPO4 5, NaNO31, MgSO4.7H2O 1; it was testified by the single factor experiment that sealing the shaking flasks with pledget other than plastic paper can obviously increase the xylanase activity; moreover, Z7 showed better xylanase production ability when in the alkali environment. Under the optional fermentation condition, the maximal xylanase activity 122.23IU/mL was proved. Through the analysis of thin- layer chromatography (TLC), the ability of xylanase from Z7, which can hydrolyze xylan to xylose monomer and oligomer, was vividly displayed.

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本文主要研究了泸州老窖古酿酒作坊内外环境空气真菌和空气细菌的群落结构和分布特征。结果如下: 作坊内外环境空气微生物浓度差别显著,并随季节变换而变化,春、夏季微生物浓度较高,秋、冬季较低,空气真菌在夏季达到最高,细菌在春季最高。 古作坊内外环境检测到的真菌均为16 属,但优势菌属不同,作坊外的优势菌属为青霉属(Penicillium)、曲霉属(Aspergillus)、无孢菌(non-sporing)、枝孢霉属(Cladosporium)和链格孢属(Alternaria);而作坊内优势菌属为曲霉属、青霉属、酵母菌(Yeast)、无孢菌,作坊内还含有较高浓度的根霉属(Rhizopus)、毛霉属(Mucor)、短梗霉属(Aureobasidiu),枝孢霉属和链格孢属等,曲霉属、酵母菌、根霉属、毛霉属为古酿酒作坊重要的酿酒真菌,青霉属、链格孢属为酿酒不利菌群。对古作坊内曲霉属进行了初步鉴定,主要是小冠曲霉(A.cristatellus)、米曲霉(A.oryzae)、黑曲霉(A.niger)和白曲霉(A.cadidus)。 空气细菌10 属21 种,作坊内外环境的优势菌属均为芽孢杆菌属(Bacillus)、微球菌属(Micrococcus)、葡萄球菌属(Staphylococcus)、假单胞菌属(Pseudomonad),其中芽孢杆菌属在作坊内占有绝对的优势,浓度比在40℅以上,是古酿酒作坊重要的酿酒细菌,另外还检测到较高浓度的乳酸杆菌(lactobucillus),这类菌容易使酒味发涩发苦,为酿酒不利菌。 作坊内外环境空气微生物表现出明显的交流现象。作坊内,青霉属、枝孢霉属、链格孢属、葡萄球菌属等杂菌占有一定比例;而在作坊外,芽孢杆菌属、曲霉属、根霉属(Rhizopus)、酵母菌等处于相对较高水平,绿化环境较好的营沟头作坊内的短梗霉属,枝孢霉属和链格孢属等杂菌含量低于什字头和新街子作坊。 The community structure and distribution characteristic of airborne microbes was investigated in ancient brewage workshops of luzhoulaojiao. The results are as follows: The concentration of airborne microbes was different in interior and exterior environment of ancient workshops, and also varied by seasons. microbial concentration was higher in spring and summer, and lower in fall and winner. The highest levels of airborne bacteria was in spring, but the fungal’s in summer. The identified genus of fungi were 16 in interior and exterior environment of the ancient workshops. But the dominant genus were different , The advantage genus in the interior were Aspergillus, Yeasts, Penicillum and Nonsporing and in the exterior were Penicillum, Nonsporing, Cladosporium, Aspergillus and Aureobasidiu. Rhizopus ,mucor, Aureobasidiu, Cladosporium, Alternaria and all also were at a higher level. Among these, Aspergillus, Yeasts, Rhizopus ,mucor are important vintage flora . Penicillum, Alternaria do harm to vintage. Aspergillus of ancient workshops was identified , the preponderant aspergillus species were A.cristatellus, A.oryzae, A.niger and A.cadidus in ancient brewage workshops. 10 genus 21 species bacteria were identified, the advantage genuses among the interior and exterior of the three workshops were bacillus, microccus, Staphylococcus Pseudomonas. Bacillus, which account for beyond 40℅ of the total bacteria concentration in all sampling pots, was the most dominant genus. Lactobacillus was identified at a high level in ancient workshops, it makes spirit taste bitter and astringent. So it is not a kind of good bacterium for vintage. The fungus in the interior and exterior atmosphere characterized intercommunion phenomenon. Obviously, the concentration of profitless fungus such as Penicillum, Cladosporium, Alternaria appeared in the interior, and the fungus such as Bacillus, Aspergillus, Rhizopus and Yeasts in the exterior were at a relatively high level. the harmfull fungus in yinggoutou workshops such as Aureobasidiu, Cladosporium, Alternaria and all were lower than shenzitou and xinjiezi workshops.

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Immobilized with PVA,sodium alginate and activated carbon,both Zoogloea sp. and Fusarium sp.strains could degrade phenanthrene and pyrene efficiently.The optimal carrier was made of 100ρ·g -1 L PVA,5 sodium alginateρ·g -1 L and 50 activated carbon ρ·g -1 L.The degradation rates of phenanthrene and pyrene in 10 days were 87.48% and 75.34% by the immobilized bacterium,37.04% and 20.85% higher than those by the free bacterium,and the rates in 15 days were 84.36% and 74.87% by the immobilized fungus,5.35% and 11.23% higher than those by the free fungus.

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研究了黄土塬区塬面不同套种作物的幼树苹果园土壤微生物的分布和酶活性。结果表明,果园土壤中微生物的分布在果树行与套种行中是不均匀的,细菌和真菌多为果树行多于套种行;土壤的过氧化氢酶,蔗糖酶,脲酶以及中性磷酸酶活性在套种小麦的果园中以套作行高于果树行,而套种烤烟则使脲酶和中性磷酸酶活性有所降低。套种豆类有利于土壤肥力的均衡发展。

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Biodeyradation has the characteristics of simple operation,low cost,and easy application.In this research,two fungus were chosen to degrade the oil in polluted soil in laboratory and fields.In the laboratory culture test with temperature controlled,the rate of oil degradation by native fungi (F) and foreign fungi (P) are 61.8% and 66.1% respectively after 50 days.In the field test the rate of oil degradation by native fungi (F) and foreign fungi (P) are 59.8% and 70.1% respectively after 150 days.

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A lignicolous fungus,Hydnochaete tabacina(Berk.& M.A.Curtis) Ryvarden,was newly recorded from China,it was found on Quercus variabilis in Beijing.An illustrated description of this species was given based on the materials related.

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A newly described pathogen,Gyrodontium sacchari,on hardwood in tropical area was reported from Yunnan Province.The genus of Gyrodontium was also the first recorded in mainland of China.An illustrated description of the fungus was given in detail based on the specimens from China.

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We report here a facile method to obtain folic acid (FA)-protected gold nanoparticles (Au NPs) by heating an aqueous solution of HAuCl4/FA in which FA acts as both the reducing and stabilizing agent. The successful formation of FA-protected Au NPs is demonstrated by UV/Vis spectroscopy, transmission electron microscopy (TEM), selected-area electron diffraction (SAED), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). ne intracellular uptake of these nanoparticles is facilitated by HeLa cells overexpressing the folate reporter, which itself is significantly inhibited by free FA in a competitive assay as quantified by inductively coupled plasma mass spectroscopy (ICP-MS). This simple one-step approach affords a new perspective for creating functional nanomaterials, and the resulting biocompatible, functional Au NPs may find some prospective applications in various biomedical fields.

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In this communication, biosynthesis of gold nanoparticles assisted by Escherichia coli DH5 alpha and its application on direct electrochemistry of hemoglobin are reported. The gold nanoparticles formed on the bacteria surface are mostly spherical. The direct electrochemistry of hemoglobin can be achieved by incorporated into the bio-nanocomposite films on a glassy carbon electrode.

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Vaterite-type YBO3:Eu3+ crystals with interesting flower and hedgehog fungus-like structures composed of nanosheets were obtained by controlled crystallization of Y2O3 and Eu2O3 in H3BO3 solutions under acidic hydrothermal (HT) conditions. Nanosheets of uniform thicknesses were formed by preferential crystal growth along the (100) crystallographic plane and specific three-dimensional structures were further developed through a homocentric growth mechanism. Optical emission measurements showed that the HT-grown nanosheet crystals exhibited a higher ratio of the emitted red-to-orange light ratio than crystals grown from solid-state reactions. The photoluminescence intensity and emission lifetimes were also studied as a function of the Eu3+ dopant concentration and the HT synthesis temperature. The effect of some additives: a chelating ligand, a surfactant and a polymer, on the YBO3:Eu3+ crystals morphology was also investigated.

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A simple, green method was developed for the synthesis of gold and silver nanoparticles by using polysaccharides as reducing/stabilizing agents. The obtained positively charged chitosan-stabilized gold nanoparticles and negatively charged heparin-stabilized silver nanoparticles were characterized with UV-vis spectroscopy and transmission electron microscopy. The results illustrated the formation of gold and silver nanoparticles inside the nanoscopic polysaccharide templates. Moreover, the morphology and size distribution of prepared gold and silver nanoparticles varied with the concentration of both the polysaccharides and the precursor metal salts.

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P>NF-kappa B is a B-cell specific transcription factor that plays crucial roles in inflammation, immunity, apoptosis, development and differentiation. In the present study, a novel NF-kappa B-like transcription factor Relish was cloned from Chinese mitten crab Eriocheir sinensis (designated as EsRelish) by rapid amplification of cDNA ends (RACE) technique based on expressed sequence tag (EST). The full-length cDNA of EsRelish was of 5034 bp, consisting of a 5' untranslated region (UTR) of 57 bp, a 3' UTR of 1335 bp with two mRNA instability motifs (ATTTA), a polyadenylation signal sequence (AATAAA) and a poly (A) tail, and an open reading frame (ORF) of 3645 bp encoding a polypeptide of 1214 amino acids with a calculated molecular mass of 134.8 kDa and a theoretical isoelectric point of 5.26. There were a typical Rel homology domain (RHD), two nuclear localization signal (NLS) sequences (KR), an inhibitor kappa B (I kappa B)-like domain with six ankyrin repeats, a PEST region and a death domain in the deduced amino acid sequence of EsRelish. Conserved domain, higher similarity with other Rel/NF-kappa Bs and phylogenetic analysis suggested that EsRelish was a member of the NF-kappa B family. Quantitative real-time RT-PCR was employed to detect the mRNA transcripts of EsRelish in different tissues and its temporal expression in hemocytes of E. sinensis challenged with Pichia methanolica and Listonella anguillarum. The EsRelish mRNA was found to be constitutively expressed in a wide range of tissues. It could be mainly detected in the hemocytes, gonad and hepatopancreas, and less degree in the gill, muscle and heart. The expression level of EsRelish mRNA in hemocytes was up-regulated from at 3, 6, 9 and 12 h after P. methanolica challenge. In L. anguillarum challenge, it was up-regulated at 9, 12 and 24 h. The results collectively indicated that EsRelish was potentially involved in the immune response against fungus and bacteria.

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本论文对民间药用红树植物木榄(Bruguiera gymnorrhiza)中的次生代谢产物进行了研究,同时还对分离自似瘤凹顶藻(Laurencia similis Namet Saito)的内生真菌EN21(Exophiala oligosperma)进行了化学成分研究。对分离得到的部分化合物进行了初步的DPPH自由基清除活性和抗菌活性筛选。 采用常规的硅胶柱层析、反相硅胶柱层析、制备薄层层析、凝胶Sephadex LH-20 柱层析、半制备型高效液相色谱(HPLC)以及重结晶等手段,从木榄(Bruguiera gymnorrhiza)中分离得到23个化合物;从EN21(Exophiala oligosperma)中分离得到23个化合物。利用一维、二维核磁、质谱、元素分析等各种现代波谱技术及化学反应方法,确定了木榄中20个化合物的结构,包括黄酮单体化合物5个,三萜类化合物11个,甾体化合物2个,长链烷烃2个,其中4个化合物为首次从木榄中报道的黄酮;确定了EN-21中20个化合物的结构,其中脑苷脂1个,甾体化合物7个,芳环化合物3个,三萜类化合物2个,包括1个新天然产物。本文系首次报道Exophiala oligosperma 的化学成分。 对从木榄中分离得到的部分化合物进行了DPPH自由基清除活性筛选, 发现黄酮类化合物BRG-1、BRG-2、BRG-3 、BRG-4显示了较强的DPPH自由基清除活性,而三萜类化合物则未表现出DPPH自由基清除活性。抗菌筛选实验中所有测试样品未显示出活性。 研究结果充实了红树林植物和海藻内生真菌代谢产物的内容,为其有效利用提供了一定的科学依据。

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由于生长环境的特殊性,红树林及其内生真菌的代谢产物在化学类型和生物活性方面都具有多样性,因此对其代谢产物的研究引起了人们越来越多的关注。本论文以菌丝体生物量、代谢产物量等指标及薄层色谱分析、高效液相色谱分析、抗菌活性测试等筛选手段对来源于我国海南红树植物的九株内生真菌在四种不同液体培养基上的静置发酵产物进行了综合评价,并从中选择了来源于半红树植物黄槿(Hibiscus tiliaceus)的内生真菌G2——赤散囊菌(Eurotium rubrum)进行了30 L规模发酵(采用PDB培养基)和次生代谢产物的研究,对分离得到的部分化合物进行了初步的生物活性评价。此外,本论文还对海南真红树植物红海榄(Rhizophora stylosa Griff.)的化学成分进行了研究,并对分离得到的部分化合物进行了二苯代苦味酰自由基(DPPH)清除活性的评价。 采用常规的硅胶柱层析、Sephadex LH-20柱层析、反相硅胶柱层析、制备薄层层析(pTLC)、重结晶等分离手段,分离纯化得到单体化合物。综合运用现代波谱技术 (IR、UV、MS、1D-NMR 和 2D-NMR) 以及与标准品或文献比对鉴定单体化合物的结构。从G2菌丝体和发酵液的合并提取物中鉴定了45个化合物的结构,其中13个为新化合物,结构类型包括6个苯甲醛类化合物(ER1*~ER6*)、4个蒽醌类化合物(ER15*~ ER18*)和3个含吲哚的二酮哌嗪生物碱类化合物(ER27*~ER29*)。 对以上分离鉴定的部分单体化合物进行了DPPH自由基清除活性、拒食杀虫活性、抗细菌活性以及体外细胞毒活性的初步评价。新化合物ER15*和三个已知化合物ER20、ER39和ER40都表现很强的DPPH自由基清除活性。化合物ER15*还表现较好的拒食杀虫活性,而化合物ER5*和ER18*不但没有杀虫活性,反而能促进幼虫的生长。所测试的化合物只有ER15*和ER18*表现出微弱的抗金黄色葡萄球菌活性。所测试的化合物对A-549、HL-60和P-388细胞株均未表现出有意义的体外细胞毒活性。 从红海榄枝条的提取物中分离鉴定了29个化合物,其中2个为新化合物,包括1个三萜酯(RS1*)和1个黄烷醇类化合物(RS12*);另有1个三萜酯(RS5)和1个黄烷醇类化合物(RS11)作为新的天然产物被分离鉴定。 对从红海榄提取物乙酸乙酯相和正丁醇相分离得到的部分单体化合物进行了DPPH自由基清除活性的研究。黄烷醇类化合物RS16和RS17显示最强的活性。另外,实验结果说明黄烷醇类化合物的DPPH自由基清除活性与其分子中所含的羟基数目有一定关系,而且若芳香环上有多个邻位酚羟基,则该化合物的活性将增强。本论文实验结果为海南红海榄植物资源的利用提供了化学成分及抗氧化活性方面的科学依据。