竹黄菌固态发酵竹红菌素条件的研究

研究了竹黄菌固体发酵培养生成竹红菌素过程中培养条件的影响。结果表明,竹黄菌生长及竹红菌素生成适宜的温度、pH分别为26℃、5．5—6。最佳的培养基物料厚度为5cm左右。固态发酵过程中最适的水份含量为50％。玉米粉是一种适宜的培养基基本组成。在以上发酵条件下竹红菌素产量为4mg/kg。     

竹黄菌液体培养下产竹红菌素的研究

先对不同产地采集的竹黄菌进行筛选,得到优产竹红菌素的菌株,然后采用单因子和3因素3水平正交试验法对竹红菌素液体发酵条件进行优化,在优化培养基的基础上,选用不同浓度的Cr3+、Fe3+、Cu2+和Ca2+对竹红菌素进行离子调控研究。结果表明:从休宁所采集的菌株不仅生长速度最快,发酵所产的竹红菌素含量也最高;竹红菌素最佳发酵碳源是葡萄糖,最佳发酵氮源是硝酸钠,最佳培养基组合为2%葡萄糖,0.2%硝酸钠,pH7.5;Cr3+和Fe3+浓度为0.005%时竹红菌素含量均最高;0.05%的Ca2+最有利于竹红菌素的分泌;Cu2+为0.03%时竹红菌素含量达到最大值。     

Study on fermentation hypocrellin by Shiraia bambusicola in submerged cultures

先对不同产地采集的竹黄菌进行筛选,得到优产竹红菌素的菌株,然后采用单因子和3因素3水平正交试验法对竹红菌素液体发酵条件进行优化,在优化培养基的基础上,选用不同浓度的Cr3+、Fe3+、Cu2+和Ca2+对竹红菌素进行离子调控研究.结果表明:从休宁所采集的菌株不仅生长速度最快,发酵所产的竹红菌素含量也最高;竹红菌素最佳发酵碳源是葡萄糖,最佳发酵氮源是硝酸钠,最佳培养基组合为2%葡萄糖,0.2%硝酸钠,pH7.5;Cr3+和Fe3+浓度为0.005%时竹红菌素含量均最高;0.05%的Ca2+最有利于竹红菌素的分泌;Cu2+为0.03%时竹红菌素含量达到最大值.     

Shiraia sp．SUPER—H168 ITS-5．8S rDNA序列分析及产竹红菌素条件初步优化

对产竹红菌素的菌株SUPER-H168的ITS-5．8S rDNA进行了序列测定和进化树聚类分析,表明其属于竹黄属;对该菌株固态发酵产竹红菌素的条件进行了初步优化,得到的初步优化条件为玉米渣25g,麸皮5g,葡萄糖1．5g,NaNO3 0．15g,ZnSO4·7H2O 0．03g,起始含水量为50％,起始pH为7．0,培养温度为30℃。在该条件下发酵18d,竹红菌素的产量为9．37mg／g干基。     

竹黄菌发酵生产竹红菌素的研究进展

竹黄作为名贵中药,其主要成分竹红菌素是优良的天然光敏药物。由于天然竹黄资源有限,通过竹黄无性型菌株发酵生产竹红菌素因而受到广泛关注。通过介绍竹黄无性型菌株的固态、液态发酵,分析了竹红菌素发酵生产的各种影响因子,并对竹红菌素合成诱导物质及高产菌株诱变筛选等进行了综述,为竹红菌素合成调控和发酵生产提供了参考。     

竹黄遗传多样性与竹红菌素合成和抗癌作用的研究进展

竹黄在我国是一种重要的药用真菌,主要活性代谢产物为竹红菌素等苝醌类光敏色素。前人关于竹黄的研究主要集中在活性成分分析、临床应用和发酵培养等方面。竹黄遗传多样性的存在是不同居群、不同菌株之间竹红菌素生物合成代谢差异的原因,并且随着竹红菌素及其衍生物对肿瘤细胞的抑制机制被不断揭示,对于竹黄的遗传多样性、竹红菌素的合成代谢与抗癌作用机理等研究正持续深入。应用不同的分子标记发现,竹黄遗传多样性既可能来源于居群内,也可能来源于居群间;而通过化学修饰或物理修饰都使竹红菌素在肿瘤的光动力治疗(Photodynamic therapy,PDT)上展现出诱人的前景。同时,影响竹红菌素产生的聚酮合酶基因等新的研究发现最终将会有助于掌握竹黄中竹红菌素的生物合成途径。综述了竹黄的遗传多样性、竹红菌素及其衍生物对癌细胞的作用和竹红菌素生物合成的研究进展,旨在为竹黄的资源保育和竹红菌素的深入研发提供参考。     

竹黄菌液体发酵条件的优化

目的：筛选并优化竹黄菌液体发酵培养基。方法：采用单因子和3因素3水平正交试验法,从生物量和竹红菌素两方面筛选碳源、氮源和pH,探讨该菌生长的最佳培养条件。结果：最佳发酵碳源是葡萄糖,最佳发酵氮源是硝酸钠。竹红菌素生长的最佳配方为2%葡萄糖,0.2%硝酸钠,pH7.5;菌丝体生长的最佳组合为2%葡萄糖,0.3%硝酸钠,pH7.5。     

Shiraia sp.SUPER-H168液态发酵产竹红菌素过程中生理特征初步研究

通过对竹黄菌株SUPER-H168产竹红菌素及不产竹红菌素时生理特征的研究,发现MDA、胞内及胞外的总抗氧化力、过氧化氢、维生素C的含量均比对照组高;CAT的活力上升,而SOD活力总体稍有下降。实验结果表明,菌体产竹红菌素初期伴随着其自身氧化水平的上升。添加终浓度为40μmol/L的过氧化氢可以促进竹红菌素生成。     

酿酒酵母菌培养条件的研究

目的：优化酿酒酵母液体发酵得到菌体的最佳条件。方法：通过单因素试验,以吸光度为指标,研究碳源、氮源、接种量、pH值及无机离子对酿酒酵母菌生长的影响。结果：酿酒酵母生长的最佳碳源是葡萄糖,最佳氮源是蛋白胨,最佳接种量2％,最佳初始pH为4．5,添加无机盐硫酸亚铁能够促进其生长。结论：得到了酿酒酵母液体生长的最佳培养基配方。     

Bei醌类化合物的杀线虫活性

采用液体浸泡方法首次测定了菌寄生菌属Hypomyces sp.的次级代谢物Bei醌类化合物竹红菌甲素和痂囊羟菌素对松材线虫的致病率，结果表明竹红菌甲素对松材线虫的半致病率为18小时50ug/mL，痂囊羟菌素对松材线虫的半致病率为18小时15ug/mL，对比（啊维菌素）对松材线虫的半致病率为18小时0．8ug/mL，在红外光照射下竹红菌甲素和痂囊羟菌素具有良好的光敏杀伤松材线虫的作用。     

紫花苜蓿多元杂交后代优良株系筛选研究

通过对甘农3号、甘农5号和游客紫花苜蓿多元杂交后代选育的36个株系及其亲本的生长、产量、品质等相关指标的测定,采用灰色关联度理论,构造综合评价模型进行供试材料综合评价,筛选出速生12#、速生11#株系为最理想的优良株系,生长高度分别为105.44cm、105.42cm;生长速度分别为1.74cm/d、1.68cm/d;茎叶比分别为0.30、0.35;分枝数分别为23、17;鲜草产量分别为39.99 t/hm2、35.13 t/hm2;粗蛋白含量分别为19.95%、23.89%;相对饲用价值为153.15%、157.02%。多叶2#、速生5#、速生20#、速生21#等4个株系为较理想的优良株系。     

原生质体紫外诱变选育竹红菌甲素高产菌株

采用原生质体紫外诱变技术选育竹红菌甲素高产菌株。结果表明:以竹黄菌Shiraia sp.S8为出发菌株,当使用混合酶系(5 mg/mL纤维素酶和10 mg/mL蜗牛酶)在30℃处理菌丝2 h,获得菌丝原生质体3.24×106个/mL。以竹黄菌原生质体在距离15 W紫外灯30 cm处照射诱导,获得诱变菌株C6。其竹红菌甲素产量达到28.1 mg/L,比原始出发菌株提高了53.7%,且遗传稳定,具有较高的医药与工业应用价值。     

Effects of the phenotypic polarization state of human leukocytes on the optical absorbance spectrum

This study evaluated the optical absorbance spectrum of human monocytes, neutrophils and lymphocytes polarized, or not, to the inflammatory or immunoregulatory phenotypes. Peripheral human blood leukocytes were isolated and polarized (10 ng/mL) with LPS or IL-4 + LPS for 2 hours. After polarization, cells were washed and incubated for an additional 24 hours (monocytes and lymphocytes) or 12 hours (neutrophils). Next, cells were collected to evaluate the optical absorbance spectrum. The three types of leukocytes exhibited absorbance in the region from 450 to 900 nm, with greater absorbance at wavelengths lower than 570 nm. Lymphocytes had a second region of greater absorbance between 770 and 900 nm. Inflammatory monocytes and lymphocytes showed increased absorbance of blue, green and yellow wavelengths (monocytes), as well as red and infrared wavelengths (monocytes and lymphocytes). Immunoregulatory polarization altered the absorbance of monocytes and lymphocytes very little. Neutrophils treated with LPS or LPS + IL-4 exhibited lower absorbance at wavelengths higher than 575 nm compared to untreated cells. The present findings showed that leukocytes exhibit greater absorbance in regions of the spectrum that have not been much used in photobiomodulation (PBM), and the polarization of these cells can affect their capacity to absorb light. Taken together, these results suggest new perspectives in the use of PBM in the clinical setting depending on the wavelengths and the stage of the inflammatory process.     

应用基因组改组技术选育竹红菌甲素高产菌株

以紫外(UV)与亚硝基胍诱变的竹黄菌(Shiraia bambusicola)菌株NU12和UV4为出发菌株,60℃处理5 min、紫外(距离30 cm,30 W)照射90s进行双亲原生质体灭活,通过30％聚乙二醇PEG6000介导原生质体融合20 min.结合平板初筛和高效液相色谱( HPLC)分析进行复筛,通过3轮重组融合操作,筛选出6株高产竹红菌甲素的融合株.其中融合菌株FIII - 21的竹红菌甲素产量达到80.4 mg/L,比原始出发菌株提高了58.9％～167.1％,且遗传稳定,具有较高的医药及工业应用价值.     

Rapid strain classification and taxa delimitation within the edible mushroom genus Pleurotus through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy

Fourier transform infrared (FT-IR) spectroscopy has been successfully applied for the identification of bacteria and yeasts, but only to a limited extent for discriminating specific groups of filamentous fungi. In the frame of this study, 73 strains - from different associated hosts/substrates and geographic regions - representing 16 taxa of the edible mushroom genus Pleurotus (Basidiomycota, Agaricales) were examined through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. A binary matrix, elaborated on the basis of presence/absence of specific absorbance peaks combined with cluster analysis, demonstrated that the spectral region 1800-600 cm(-1) permitted clear delimitation of individual strains into Pleurotus species. In addition, closely related species (e.g., Pleurotus ostreatus and Pleurotus pulmonarius) or taxa of the subgenus Coremiopleurotus demonstrated high similarity in their absorbance patterns, whereas genetically distinct entities such as Pleurotus dryinus, Pleurotus djamor, and Pleurotus eryngii provided spectra with noteworthy differences. When specific regions (1800-1700, 1360-1285, 1125-1068, and 950-650 cm(-1)) were evaluated in respect to the absorbance values demonstrated by individual strains, it was evidenced that this methodology could be eventually exploited for the identification of unknown Pleurotus specimens with a stepwise process and with the aid of a dichotomous key developed for this purpose. Moreover, it was shown that the nature of original fungal material examined (mycelium, basidiomata, and basidiospores) had an effect on the outcome of such analyses, and so did the use of different mycelium growth substrates. In conclusion, application of FT-IR spectroscopy provided a fast, reliable, and cost-efficient solution for the classification of pure cultures from closely related mushroom species.     

Molecular identification of Paragonimus ohirai and P. westermani from Anhui Province, China

Nucleotide sequences of the internal transcribed spacer 2 (ITS2) region were determined from seven adults of species Paragonimus collected from Jinde and Xiuning Counties, Anhui Province, China. Among these, the nucleotide sequence obtained from one Paragonimus adult (Jinde County) was identical to the ITS2 sequence of P. ohirai previously reported. In order to confirm the result, partial regions of mitochondrial cytochrome C oxidase I (COI) and NADH dehydrogenase 1 (ND1) from the putative P. ohirai sample were further sequenced. They showed a high level of similarity with those of P. ohirai, COI (99.7%) and ND1 (99.5%), supporting the result obtained from the ITS2. In addition to this, we designed P. ohirai- and P. westermani-specific primers (BDW and BD2OH) from ITS2 to identify P. westermani and P. ohirai easily and rapidly. After testing utility of the primers, they were applied to identify seven unidentified Paragonimus samples collected from Jinde and Xiuning Counties, China. All the examined samples showed P. westermani band pattern, and it was reconfirmed by sequencing their ITS2 regions that they are P. westermani. This result indicates that the two newly designed specific primers could be quite helpful for easily identifying P. westermani and P. ohirai, that most of Paragonimus in Jinde and Xiuning Counties consist of P. westermani, and that P. ohirai exists in Jinde County with minority.     

Depth distribution and biological characteristics of the European eel Anguilla anguilla in Lough Ennell, Ireland

Using a longline survey, a total of 196 European eels Anguilla anguilla were collected at different depths in Lough Ennell (maximum depth 30 m), central Ireland. The catch per unit of effort of A. anguilla that were caught from 1 to 25 m depths was lowest at 0·5–5·0 m and greatest at the deepest depth range (22·5–25·0 m). Sub-samples of A. anguilla from depths of <15 m showed little or no difference in size, sex ratio, age, growth rate, condition factor, length–mass relationship, gonado-somatic index, fin index or eye index with fish from depths of >15 m. All fish examined were female yellow-phase A. anguilla that had ages from 7 to 20 years (mean ± s . d . = 10·3 ± 2·9 years), with growth rates from 24·0–60·8 mm year⁻¹ (mean ± s . d . = 40·7 ± 8·5 mm year⁻¹). Variations in the growth rates were greater in the shallow group than that of the deep group. This study suggested that deeper regions are important feeding habitats for A. anguilla and that fish in this lake were growing moderately fast compared to similar habitats and areas in the species' range.     

施肥深度对矮化苹果15N尿素吸收、利用及损失的影响

以5年生烟富3/M26/平邑甜茶为试材,采用¹⁵N同位素示踪技术,研究表层(0 cm)、上层(20 cm)和中层(40 cm)3个施肥深度对矮化苹果¹⁵N 尿素吸收、分配及利用特性的影响.结果表明： 20 cm施肥处理的叶面积、叶绿素含量和叶片全氮含量显著高于0和40 cm施肥处理.不同施肥处理各器官从肥料中吸收分配到的¹⁵N量对该器官全氮量的贡献率(Ndff)存在显著差异,盛花期均以根的Ndff最高,多年生枝次之;新梢旺长期和花芽分化期根部吸收的¹⁵N优先向新生营养器官转运;果实膨大期各器官Ndff均达到较高水平;果实成熟期均以果实中的Ndff最高.果实成熟期不同施肥处理的¹⁵N分配率存在显著差异,20 cm施肥处理生殖器官和营养器官的¹⁵N分配率显著高于0和40 cm施肥处理,而贮藏器官的¹⁵N分配率显著低于0和40 cm施肥处理.在果实成熟期,20 cm施肥处理¹⁵N肥料利用率为24.0%,显著高于0 cm(14.1%)和40 cm施肥处理(7.6%),而¹⁵N损失率为54.0%,显著低于0 cm(67.8%)和40 cm施肥处理(63.5%).不同施肥深度土壤¹⁵N残留量随施肥深度的增加而显著增加.     

Estimating cell concentration in the presence of suspended solids: a light scatter technique

A novel sensor was developed, based on light scatter, to estimate the cell concentration in the presence of suspended solids. The light scatter properties of cells in the presence of suspended solids were investigated. Two crucial observations were made: first, that the light scatter from cells is essentially a linear function of cell concentration and, second, that invariant regions are present in the light scatter spectrum of cell/solid substrate mixtures. Invariant regions are wavelength intervals of the light scatter spectrum in which the light scatter reading is independent of solid substrate concentration and only a function of cell concentration. The occurrence of invariant regions is the key behavior which allowed the quantification of cell concentration in the presence of suspended solids.An algorithm was developed for the estimation, from light scatter data, of cell concentration in the presence of solid substrate. The light scatter approach was validated by comparing cell concentrations estimated by this technique to those obtained from DNA and carbon dioxide evolution rate measurements during a series of fermentations. The model system used was Bacillus subtilis var sakainensis ATCC 21394 growing on fishmeal as the sole nitrogen source.A model was developed based on the interactions of scatter and absorbance. This model reflects the hypothesis that invariant regions are caused by changes in the absorbance of the solid substrate as a function of wavelength. (c) 1992 John Wiley & Sons, Inc.