蝉拟青霉菌丝体与天然蝉花中化学成分的比较分析

以天然蝉花Cordyceps cicadae做对比,对一株蝉拟青霉Paecilomyces cicadae菌株发酵菌丝体的化学成分进行了分析,包括粗成分、生物活性成分、无机元素、氨基酸、脂肪酸等。结果表明:蝉拟青霉菌丝体中虫草多糖的含量为33.2mg/g,甘露醇的含量为78.9mg/g,麦角甾醇的含量为0.6256mg/g,腺苷的含量为1.0620mg/g,钙、铁、锌、硒和锰微量元素的含量分别为5187.59μg/g、207.23μg/g、41.93μg/g、0.087μg/g和28.24μg/g,18种氨基酸齐全,不饱和脂肪酸是优势脂肪酸,以亚麻油酸、油酸为主,其相对含量分别为53.91%和20.63%,与天然蝉花相比,发酵菌丝体中虫草多糖、甘露醇、麦角甾醇、腺苷、氨基酸总量、必需氨基酸总量和不饱和脂肪酸的含量明显高于后者,其它化学成分和天然蝉花中相应成分基本一致。     

冬虫夏草(Cordyceps sinensis)菌丝体固体发酵粉化学成分的分析

本研究对一种冬虫夏草(Cordyceps sinensis)菌丝体固体发酵粉的化学成份进行了分析,包括粗成份,生物活性成份,脂肪酸,氨基酸,金属元素等。冬虫夏草菌丝体固体发酵粉中3’-脱氧腺苷含量40．81μg/g,腺苷含量745．4μg/g;甘露醇的含量为165．8mg／g,多糖含量为157．3mg／g;谷氨酸,天门冬氨酸,苯丙氨酸是三大类主要氨基酸;不饱和脂肪酸是优势脂肪酸,以亚油酸,油酸为主。和文献记录的结果相比,虫草发酵粉中3’-脱氧腺苷,甘露醇的含量明显高于冬虫夏草子实体中相应成份的含量;各种氨基酸的含量和水平与冬虫夏草子实体中相应氨基酸的含量和水平相当;虫草发酵粉中的其它化学成份和冬虫夏草子实体中相应成份基本一致。     

蝙蝠蛾拟青霉菌丝体化学成分的研究

应用梯度提取、反复重结晶及硅胶色谱法分离和纯化,采用NMR等谱学方法鉴定结构对蝙蝠蛾拟青霉(Paecilomyces hepiali)菌丝体进行了化学成分的分析。从蝙蝠蛾拟青霉菌丝体中分离得到6个化合物,分别鉴定为4,6,8(14),22(23)-四烯-3-酮-麦角甾烷(1)、麦角甾醇(2)、β-谷甾醇(3)、麦角甾醇过氧化物(4)、啤酒甾醇(5)、甘露醇(6)和1-油酰基-2-亚油酸-3-棕榈酸甘油(7),并对其石油醚提取物中的油状组分进行GC-MS分析,结果表明其中主要为亚油酸(9,12-十八碳二烯酸,含量41.949%)、反油酸(反9-十八碳烯酸,含量为27.696%);同时采用高效液相色谱法测定其中的麦角甾醇的含量,与海鲜菇、杏鲍菇、香菇、金针菇、滑子蘑、平菇等食用菌子实体比较,结果蝙蝠蛾拟青霉菌丝体中麦角甾醇含量为15.65 mg/g,平菇为21.43 mg/g,杏鲍菇为10.16 mg/g,香菇为13.34 mg/g,金针菇为9.48 mg/g,滑子蘑为10.43 mg/g,海鲜菇为14.42 mg/g。     

蝉虫草子实体与发酵菌丝体胞内多糖成分分析

比较蝉虫草子实体、孢梗束、虫体以及蝉虫草发酵菌丝体的胞内多糖含量、单糖组成和分子量差异,结果表明,蝉虫草子实体与发酵菌丝中胞内多糖的含量和组成均存在差异。葡萄糖、半乳糖和甘露糖是蝉虫草发酵菌丝体和子实体粗多糖中主要的3种单糖,阿拉伯糖和木糖是子实体粗多糖中的特征性单糖,而果糖属于菌丝体的特征性单糖。子实体粗多糖中高分子量组分（>1×10 ⁶Da）的相对含量明显高于菌丝体粗多糖。本研究对提升蝉虫草发酵多糖产品品质、促进蝉虫草开发应用具有参考价值。     

灵芝孢子粉中脂溶性成分分析方法的建立

运用高效液相建立灵芝孢子粉中脂溶性成分的分析测定方法。通过色谱柱、洗脱条件、ELSD参数的优化,建立了12种脂溶性成分的测定方法,结果表明,该方法简单、准确、稳定,可以实现孢子粉中甘油三酯、脂肪酸、甾醇三类脂溶性成分的同时提取、分析;破壁孢子粉中脂溶性成分为301.49-397.37mg/g,孢子油产品中脂溶性成分的含量为626.00-713.07mg/g,远高于三萜含量;1,2-二油酸-3-棕榈酸甘油酯、甘油三油酸酯、1,2-二油酸-3-亚油酸甘油酯是主要的脂溶性成分,脂肪酸以不饱和脂肪酸亚油酸及油酸为主,甾醇中麦角甾醇含量最高。研究结果明确了灵芝孢子粉中脂溶性成分的物质基础,为深入研究其活性成分、全面评价孢子粉质量提供了依据。     

蝙蝠蛾拟青霉液体发酵工艺优化及菌丝体腺苷含量的检测

通过摇瓶液体培养的方法,以菌丝体生物量为主要考察指标,筛选蝙蝠蛾拟青霉PH-2菌株液体发酵的培养基成分和条件。进一步采用正交试验,以菌丝体生物量和腺苷含量为联合指标,确定PH-2菌株液体发酵高产菌丝体和腺苷的培养基配方。通过试验获得的最佳配方为：玉米淀粉40g/L,葡萄糖5g/L,玉米浆25g/L,硫酸镁·7H₂O 1g/L,磷酸二氢钾1g/L,硫酸锌1.5g/L,维生素B1 25mg/L;培养条件为：初始pH 6.0,装液量150mL/500mL,接种量8%（V/V）,温度26℃,转速120r/min,培养7d。在该工艺下,菌丝体生物量达到25.2g/L。应用高效液相色谱（HPLC）法,检测到菌丝体中腺苷含量为2.76mg/g。该方法简便、快速、准确,可作为蝙蝠蛾拟青霉菌丝体或其相关产品中腺苷的检测手段。本工艺的优化试验获得了较高的菌丝体收率,菌丝体质量良好。     

不同干燥方式对培养蝉花孢梗束品质的影响

本研究为探究不同干燥方式对蝉花孢梗束品质的影响和为生产中蝉花孢梗束选择合适的干燥方式提供理论依据,研究了自然干燥、热风干燥、微波干燥、热泵干燥和冷冻干燥对蝉花孢梗束粉的外观、粒度、主要营养和功能成分的影响。运用主成分和聚类分析对不同干燥处理得到的样品的数据进行综合分析。研究表明热泵干燥能最好地保持样品的亮黄色色泽,蓝黄度b*为36.74,显著高于其他方式干燥样品,其次是微波干燥;但是这两种方式干燥的蝉花孢梗束总氨基酸和必需氨基酸含量均较低。自然干燥和冷冻干燥的样品感观较差,蓝黄度b*均小于28且二者差异不显著。热风干燥样品色泽仅次于微波干燥,含有较高蛋白质、总氨基酸和总糖,且其脂肪、麦角甾醇和甘露醇的含量最高,分别为4.15 g/100 g、8.65 mg/g和59.20 mg/g。总体考虑,热风干燥能较好地保持蝉花孢梗束的色泽、营养和功能成分,且技术成熟度高,适用于工业化大生产。     

蝙蝠蛾拟青霉发酵滤液的成分组成与急性毒性分析

蝙蝠蛾拟青霉是从新鲜的冬虫夏草上分离获得的虫草菌,可采用生物发酵制备大量菌丝体。该菌丝体具有和冬虫夏草相似的活性成分,具有抗肿瘤、抗病毒等多种功效。本研究对干燥后的蝙蝠蛾拟青霉发酵滤液的主要成分进行了分析,结果显示：干燥后的蝙蝠蛾拟青霉发酵滤液含粗脂肪21.33%,总蛋白27.34%,水解氨基酸16.79%,粗多糖11.16%,不饱和脂肪酸71.21%,其中以油酸、亚油酸为主;该干燥发酵滤液中还含有多种核苷类物质、多糖等生物活性成分以及多种人体必需的矿质元素。此外急性毒性实验的初步结果表明：小鼠在实验期内无死亡现象和明显的中毒反应,主要脏器的形态学正常;谷草转氨酶（AST）和谷丙转氨酶（ALT）活性与对照组相比无统计学差异。蝙蝠蛾拟青霉发酵滤液有望在医药和保健领域进一步研究开发。     

4种虫草相关真菌菌丝体粗多糖的生物活性评价

本研究以细脚棒束孢、蛹虫草、蝉棒束孢和球孢白僵菌的菌丝体粗多糖为对象,分析4种虫草相关真菌菌丝体粗多糖含量与生物量的相关性,并进一步对其抗氧化能力和抗肿瘤活力进行评价.液体发酵结果表明,蝉棒束孢MF12、MF13和蛹虫草MF27、MF1的菌丝体粗多糖含量(＞40mg/g)显著高于其他菌株,蝉棒束孢MF11、MF13和蛹...     

不同蝉花产品蛋白质营养价值分析

采用国际通用的营养价值评价方法 FAO/WHO,对野生蝉花、蝉花液体发酵菌丝体、蝉花固体培养孢梗束和菌质的蛋白质的营养价值进行了全面评价。结果显示,其蛋白质含量分别为19.65%、25.30%、32.90%和16.17%。野生蝉花和蝉花液体发酵菌丝体的第一限制氨基酸是含硫氨基酸(蛋氨酸和半胱氨酸),蝉花孢梗束的第一限制氨基酸是异亮氨酸,蝉花菌质的第一限制氨基酸是赖氨酸。综合评价,蝉花孢梗束蛋白质的营养价值最高,蝉花菌质蛋白质的氨基酸平衡性较好。     

蝉拟青霉液体发酵条件优化

采用液体发酵蝉拟青霉,对蝉拟青霉的发酵条件进行优化,以提高蝉拟青霉胞外多糖产量及生物量。摇瓶发酵条件下,在单因素基础上设计正交实验确定各因素的最佳组合。优化后得最佳发酵培养基:蔗糖8%,牛肉膏0.75%,酵母膏0.125%,MgSO_4·7H_2O 0.3%,KH_2PO_4 0.2%,麸皮0.5%。该条件下胞外多糖产量为5.96 g/L,生物量为42 g/L,较优化前提高了1倍。采用发酵罐进行扩大培养,对分批发酵时的初糖浓度进行了优化,并分析了补料分批发酵对发酵过程的影响。发酵罐培养时最适初糖浓度为5%,此时生物量最高为38 g/L,多糖含量最高为5.5 g/L;采用补料分批发酵时,多糖产量最高为5.89 g/L,生物量最高为40 g/L,效果优于分批发酵。     

贵阳地区蝉花微生态系统中细菌种群结构与功能

[目的]探索自然生态下蝉花内菌核、菌膜和环境细菌群落结构、功能及其相互关系.[方法]对细菌16SrRNA扩增片段进行高通量测序,分析贵阳市大将山和贵阳森林公园的蝉花内菌核、菌膜及其生境土壤的细菌群落组成、多样性及潜在功能.[结果]蝉花内菌核样本共检测到细菌562个属,菌膜样本521个属,菌际土样本578个属.两地的各组...     

蝉棒束孢显微形态变异式样

【目的】揭示蝉棒束孢居群形态性状的变异式样。【方法】通过对15个居群,75个蝉棒束孢菌株17个显微形态性状的观察和测量,采用SAS 8.1软件对蝉棒束孢显微形态性状数据进行描述性、巢式方差和Q聚类分析,获得蝉棒束孢显微形态变异数据。【结果】蝉棒束孢具有大小两型分生孢子,产孢细胞的形态可分为葫芦状和瓶状2种类型;蝉棒束孢在PDA培养基中易形成数量众多的厚垣孢子,菌丝间多能形成融合菌丝,并可观察到少量的融合菌丝在厚垣孢子与菌丝间、产孢细胞间形成。描述性分析表明,蝉棒束孢居群17个形态性状的变异系数(CV)在13.07%–104.09%之间,在物种水平上表现出丰富的变异;巢式方差分析表明,蝉棒束孢显微形态性状在居群间平均方差分量占总变异的11.29%,居群内个体间平均方差分量占总变异的15.27%,菌株内观察值间平均方差分量占总变异的73.44%。【结论】蝉棒束孢菌株形态性状变异丰富,是形态性状变异的主要来源,蝉棒束孢居群显微形态性状不存在明显的地理相关性。     

引致蝉若虫地方病的蝉棒束孢种群遗传结构

采用ISSR分子标记技术对在3个不同地区引起蝉若虫地方病的蝉棒束孢种群遗传结构进行研究.结果表明: 3个地方病种群均表现出较高的遗传多样性,其中敬亭山种群的遗传多样性最高,石台种群最小.UPGMA聚类分析表明,不同地方病种群无优势性,均为多系的异质种群,遗传谱系与地理来源无关.敬亭山不同采集时间的菌株表现出明显的时间异质性.种群(亚种群)间的遗传分化系数G_st为0.2153,而基因流N_m为0.9110(N_m＜1), 暗示低水平的基因交流是种群内遗传变异的主要原因之一.因此,蝉棒束孢种群水平的高异质性和低优势性是维持蝉若虫地方病的遗传结构特征.     

Effects of MULTIFOLIATE-PINNA, AFILA, TENDRIL-LESS and UNIFOLIATA genes on leafblade architecture in Pisum sativum

In order to dissect the genetic regulation of leafblade morphogenesis, 16 genotypes of pea, constructed by combining the wild-type and mutant alleles of MFP, AF, TL and UNI genes, were quantitatively phenotyped. The morphological features of the three domains of leafblades of four genotypes, unknown earlier, were described. All the genotypes were found to differ in leafblade morphology. It was evident that MFP and TL functions acted as repressor of pinna ramification, in the distal domain. These functions, with and without interaction with UNI, also repressed the ramification of proximal pinnae in the absence of AF function. The expression of MFP and TL required UNI function. AF function was found to control leafblade architecture multifariously. The earlier identified role of AF as a repressor of UNI in the proximal domain was confirmed. Negative control of AF on the UNI-dependent pinna ramification in the distal domain was revealed. It was found that AF establishes a boundary between proximal and distal domains and activates formation of leaflet pinnae in the proximal domain.     

Presence of Rhizobium Etli bv . Phaseoli and Rhizobium Gallicum bv . Gallicum in Egyptian Soils

Seven bean rhizobial strains EBRI 2, 3, 21, 24, 26, 27 and 29 identified as Rhizobium etli, and EBRI 32 identified as Rhizobium gallicum, isolated from Egyptian soils and which nodulated Phaseolus vulgaris efficiently, were subjected to hybridization with a nifH probe in order to estimate the copy number of this gene. Seven strains (EBRI 2, 3, 21, 24, 26, 27 and 29) which were only able to nodulate Phaseolus vulgaris, contained three copies of the nifH gene, consistent with their identification as Rhizobium etli bv. phaseoli. Only one strain (EBRI 32) which nodulated both Phaseolus vulgaris and Leucaena leucocephala, had one copy of nifH gene. This confirmed the classification of this strain as Rhizobium gallicum bv. gallicum.     

Characterisation of cell wall polysaccharides, arabinogalactans-proteins (AGPs) and phenolics of Cola nitida, Cola acuminata and Garcinia kola seeds

Many Cola plant species are endemic to West and Central Africa. Cola acuminata and Cola nitida are used as masticatory when fresh, while the dried nuts are used for beverages and pharmaceutical purposes in Europe and North America. Garcinia kola seeds, that serve as a substitute for the true kola nuts, are used in African traditional medicine for the treatment of various diseases, including colic, headache and liver cirrhosis. Seeds extracts of G. kola are also known for their anti-inflammatory, antimicrobial and antiviral properties. To gain information on the chemical properties of the kolas, we have isolated and analyzed cell wall polysaccharides, arabinogalactan-proteins and phenolic substances from the seeds of the three kola species. The sugar composition of cell wall material of C. acuminata, C. nitida and G. kola revealed that Gal (up to 30%), Ara, GalA and Glc as the predominant monosaccharides, representing approximately 90% by mol of the total hydrolysable sugar present in this material. In Ammonium oxalate cell wall fraction, GalA was found to be the major sugar present in all kola species. In the alkali-soluble fraction, there were significant differences in the level of Glc and Gal. The level of Glc was high in C. acuminata and C. nitida while the level of Gal and Xyl were high in C. nitida and G. cola. Isolation and quantification of arabinogalactan-proteins demonstrate that G. kola seeds contained four to eight times more of these proteoglycans than the seeds of the other two species. Finally, analysis of soluble phenolic substances shows that caffeine and catechin were largely represented in C. acumina and C. nitida seeds, with caffeine accounting for 50% of all soluble phenolics. These findings indicate that the three Kola seeds are highly enriched in pectins and proteoglycans and that C. acuminata and C. nitida can be used as a possible source of caffeine and catechin.