Isaria cicadae Miquel is an important medicinal food fungus. The crude polysaccharides from I. cicadae were shown to significantly extend the lifespan of fruit flies in our previous study. To understand the underlying mechanism, the composition and structural characterization of polysaccharides from I. cicadae were determined. A water-soluble crude polysaccharide, obtained from a cultured Isaria cicadae by boiling-water extraction and ethanol precipitation, was fractionated by DEAE-Sepharose column chromatography. The isolate gave four polysaccharide fractions termed B-I-1, B-I-2, B-II-1 and B-II-2. The average ratio of Glu: Man: Gla in B-I-2 was 1.28:1.25:1, B-II-1 was 16.08:1:1.95, and B-II-2, was 1:2.43:2.61. B-I-1 was composed of Ara, Xyl and Glu in the ratio of 1:2.36:33.14. GC and FTIR spectra analysis combined with NMR spectroscopic analysis gave preliminary structures of the four polysaccharide fractions, which may provide an elucidation on the action mechanisms of - polysaccharide on fruit flies proliferation.

Mice were divided into one negative group (given 1％ sodium carboxymethyl cellulose) and three treatment groups at different doses,i.e.,low-dose,middle-dose and high-dose groups.Mice were fed with Chanhuabao tablets by five,ten and thirty times of the dose recommended to human beings for 4 weeks,respectively.The abilities of natural killer (NK) cells were tested.The delayed type hypersensitivity (DTH) test and splenic lymphocyte proliferation test were carried out thereafter.The results showed that compared to the negative group,the three treatment groups showed significant swelling of mice (P＜0.01).The level of serum hemolysin (P＜0.01),hemolytic plaque number (P＜0.01) as well as the phagocytosis rate and phagocytic index (P＜0.01) were significantly improved in the middle-and high-dose groups.The killing activity of NK cell was significantly increased in the high-dose group (P＜0.05).In conclusion,Chanhuabao tablet could significantly improve the immunological function of mice.

为了了解红豆杉Taxus chinensis内生真菌的种类及分布特点,通过采样、分离、培养与鉴定,对安徽黄山红豆杉内生真菌的群落结构及生态分布进行了研究.结果共分离出14属107株内生真菌,其中青霉属Penicillium、曲霉属Aspergillus、交链孢属Alternaria和拟青霉属Paecilomyces为优势种群.同时内生真菌的多样性具有组织专一性,以根和皮中分布较多,分别是分离菌株总数的37.38％和26.17％.

The entomopathogenic fungus, Beauveria bassiana, is commonly used as a biological agent for pest control. Environmental and biological factors expose the fungus to oxidative stress; as a result, B. bassiana has adopted a number of anti-oxidant mechanisms. In this study, we investigated metabolites of B. bassiana that are formed in response to oxidative stress from hydrogen peroxide (H2O2) by using a liquid chromatography mass spectrometry (LC-MS) approach. Partial least-squares discriminant analysis (PLS-DA) revealed differences between the control and the H2O2-treated groups. Hierarchical cluster analysis (HCA) showed 18 up-regulated metabolites and 25 down-regulated metabolites in the H2O2-treated fungus. Pathway analysis indicated that B. bassiana may be able to alleviate oxidative stress by enhancing lipid catabolism and glycometabolism, thus decreasing membrane polarity and preventing polar H2O2 or ROS from permeating into fungal cells and protecting cells against oxidative injury. Meanwhile, most of the unsaturated fatty acids that are derived from glycerophospholipids hydrolysis can convert into oxylipins through autoxidation, which can prevent the reactive oxygen of H2O2 from attacking important macromolecules of the fungus. Results showed also that H2O2 treatment can enhance mycotoxins production which implies that oxidative stress may be able to increase the virulence of the fungus. In comparison to the control group, citric acid and UDP-N-acetylglucosamine were down-regulated, which suggested that metabolic flux was occurring to the TCA cycle and enhancing carbohydrate metabolism. The findings from this study will contribute to the understanding of how the molecular mechanisms of fungus respond to environmental and biological stress factors as well as how the manipulation of such metabolisms may lead to selection of more effective fungal strains for pest control.

To observe the effects of synnema and mycelial of Isaria cicadae improve the role of sleep in mice , Kunming mice were randomly divided into 8 groups , 10 for every group . Per day orally given different doses of synnema and mycelial of Isaria cicadae , after continu-ous administration of the test substance of 30 days, the experiments were performed , inclid-ing direct effects of sleep test , extend pentobarbital sleep time experiment , direct sleep threshold dose of pentobarbital sodium pentobarbital sodium hypnotic sleep latency test and mice experimentally determined time , pentobarbital sodium -induced sleep time subthresh-old dose of sleep and the incidence of sodium pentobarbital -induced sleep latency .Results indicated that compared with control group , the group synnema of Isaria cicadae ( 300 mg/kg;900 mg/kg) and mycelial of Isaria cicadae (900 mg/kg) certain role for subthreshold dose of pentobarbital sodium -induced sleep in mice ( P<0 .05 ) and no clear dose -de-pendent manner .The control compared with control group , the group of synnema of Isaria ci-cadae (300 mg/kg;900 mg/kg) and mycelial of Isaria cicadae (900 mg/kg) pentobarbital induced sleep time in mice significantly prolonged action .However, two different doses of&nbsp;the test drug sodium pentobarbital no effect on sleep latency , and there were no direct role to sleep .The synnema and mycelial of Isaria cicadae have some improvement on mice sleep .

对不同品种的玉米种子及不同地区、不同部位的玉米内生真菌进行分离和鉴定.结果显示,8个不同品种的玉米种子,内生真菌检出率为25％,不同玉米品种的种子内所含的内生真菌种类和数目都不同,最高的检出率为40％,最低的检出率为10％.从玉米种子中分离得到的92个菌株,经形态学鉴定,分别属于14个属,出现频率最高的是木霉属,为34.78％.从不同地区的玉米组织中一共分离得到内生真菌168株,其中10株未产孢的没有鉴定,其他菌株经形态学鉴定分别属于26个属,以青霉属、曲霉属、头孢霉属为主,占内生真菌分离物总数的41.07％.对形态学鉴定为白僵菌属的10个菌株进行分子鉴定,均确定为球孢白僵菌(Beauveria bassiana).此外,球孢白僵菌作为玉米的内生真菌,在国内还是首次分离得到.

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.

The strong scavenging activity of metabolites in inonotus obliquus were investigated through the free radical scavenging assay in this study. At the same time,the activing ingredients LWZc and LWZf were separated and purified with MPLC,SephadexLH-20,silica gel column chromatography,secondary crystal and so on. Two pure compounds Lwzc and Lwzf were obtained. Meanwhile,their purity was tested with TLC, 1H-NMR and HPLC methods. These pure compounds were submitted to the structural analysis with thick solution HPLC-TOFMS and1H-NMR. The structure identification result revealed that the molecular formula of LWZf and LWZc were C7H6O3and C13H17N1O6respectively. The structure of LWZf was identified as 4-hydroxy Benzoic acid. Though 4-hydroxy Benzoic acid was a known compound of strong anti-oxidant activity,it was the first time to isolated the compound from. inonotus obliquus. The probably new structure of LWZc was deduced as 3-acetyl-4-amido-phenyl-ribotide,which activity was not high.

The technology of extracting crude polysaccharide from synnemata of Cordyceps cicadae culture was studied using single factor and orthogonal experiments. The biological activities of the crude polysaccharide were measured via the effect of polysaccharide on the lifespan of fruit flies and the SOD activity and MDA content in fruit flies. The results indicated that optimal extraction conditions included water to sample ratio at 20 ∶ 1,extraction time 2 h at 90℃ for twice,and precipitated by 70% ethanol for 24 h. The yield of crude polysaccharide was 8. 65%. The results demonstrated the crude polysaccharide could remarkably prolong the lifespan of female and male fruit flies for 18. 78% and26. 23%,respectively,compared with the average lifespan of their control groups. In addition,the crude polysaccharide could enhance the activity of SOD and reduce the MDA content in fruit flies.

从黄山地区红豆杉中分离得到107株内生真菌,利用薄层层析(TLC)方法对107株内生真菌的发酵代谢物进行了初筛,首次筛出8株可产紫杉醇或其类似物的菌株.再通过高效液相色谱(HPLC)对其作了进一步分析,发现有1株内生真菌菌株发酵代谢物的吸收峰与紫杉醇标准品吸收峰保留时间一致.同时结合以中国仓鼠卵巢CHO细胞株作为肿瘤细胞模型,用Resazurin法检测生长抑制率,对经筛选出的1株内生真菌次生代谢产物进行体外抗肿瘤活性试验,该菌株的代谢物对CHO细胞的抑制率高达71.28％.通过对该菌株的显微形态观察,从菌丝、孢子的形态和产孢子的特征等初步判定HQ-24是曲霉(Aspergillus sp.).

According to the extraction effects of different solvents,the physical and chemical characters tests of cordycepin,the optimal extraction condition was determined as follows:(1) centrifuging the fermenting matter with centrifugal machine and collecting the mycelium to dry by hot air at 50 ℃.(2) degreasing the fungus powder with petroleum ether according to fungus powder:the benzine=1:1's quantity.(3) using 75% ethyl alcohol,45 ℃ supersonic extracting cordycepin,and the extraction run four times with the raw material-water ratio at 1/20 and it lasted 40 minutes each time.

The volatile components in mycelia of Cordyceps formosana cultured under different conditions and artificial culture of fruiting bodies of C.formosana were extracted by simultaneous distillation and extraction(SDE) for the first time and analyzed with gas chromatography-mass spectrometry(GC-MS) in this paper.The results showed that culture conditions had great influence on the content and number of volatile components,but little impact on compound classes which mainly were alcohols,olefins,esters,phenols and ethers.There were 19 and 12 volatile components identified in mycelia of liquid and solid culture with the highest content of 1-octene-3-alcohl(51.63%) and 2-decene-5-lactone(41.13%),respectively.Fifteen compounds are identified from artificial culture of fruiting bodies,wherein the content of 1-octene-3-alcohl was highest(37.70%).The common volatile components of 3 kinds of artificial culture of C.formosana were 1-octene-3-alcohol,2-decene-5-lactone,butylated hydroxytoluene and 3-tert-Butyl-4-hydroxyanisole.This paper will provide experimental foundation and scientific basis for further development of C.formosana.

The volatile components of mycelia of Paecilomyces hepiali and Paecilomyces gunnii were extracted by simultaneous distillation extraction(SDE) and analyzed by gas chromatography-mass spectrometry(GC-MS).The results indicated that there were 25 and 32 volatile components in mycelia of P.hepiali and P.gunnii,respectively.Butylated hydroxytoluene and 2H-pyrrol-2-one,1,5-dihydro-1-methyl are the two major volatile components in the mycelia of P.hepiali,accounting to 65.78% and 12.77%,respectively.Butylated hydroxytoluene and(E,E)-2,4-decadienal are the two major volatile components in the mycelia of P.gunnii,accounting to 62.11% and 12.32%,respectively.Eight components are in common in liquid submerged fermentation mycelia of the two fungi and make up 71.58% and 69.67% of the total volatile components,respectively,indicating that the major volatile components in the two experimental mycelia are similar.The analysis of volatile components provided a better understanding of the pharmaceutical activities of the mycelia of P.hepiali and P.gunnii.

The fruiting bodies of Ophiocordyceps longissima were cultured successfully using the strain of Hirsutella longissima RCEF3891,the anamorph of O.longissima,and the effect of different growth factors on fruiting bodies formation was studied in this paper.The results showed that the optimal culture conditions for fruiting bodies of O.longissima were as follows: 9 d of the liquid fermentation period of H.longissima,with addi-tion of 10% animal protein in the medium,pH 6.0 of the nutrient solution,cultivation at 25℃,mycelia culture stage under the condition of complete darkness and continuous light(1-200 lx) after covering with mycelia.

[Objective]To select antagonistic strains against rice sheath blight and adapted to rice straw retention.[Method]Straw degradability of 4 bacterial strains of Bacillus and 9 Trichoderma strains isolated from soils were studied.Antagonistic ability of Bacillus B4 and Trichoderma T3,T5,T6 and T9,which showed better straw degradability,against rice sheath blight and adaptability to rice straws were tested.[Result]Bacillus B4 and Trichoderma T3,T5,T6 and T9 had much better degradative efficacy than other strains,of which Trichoderma T5 and T9 showed the best degradability.The results showed that the inhibition rate of Bacillus B4 was 40.48% against rice sheath blight on dual culture,and inhibition zones could maintain stable.Trichoderma T3,T5,T6 and T9 had higher inhibition rates,which were 46.48%,45.63%,50.99% and 53.38%,respectively.The metabolites from the 5 strains could inhibit the growth of rice sheath blight and completely prevent the formation of sclerotium,and the 5 strains had stronger adaptability to rice straws.[Conclusion]Trichoderma T5 and T9 had not only better straw degradability,but also stronger inhibition to rice sheath blight.

Phytases are a group of enzymes capable of releasing phosphate from phytate,and have been ap-plied mainly to animal(swine and poultry) diets in order to improve phytate P digestibility and mineral bioavail-ability.RCEF4908,a strain with high phytase yield,was screened from a lot of phytase-producted fungal isolates.According to morphological characters and internal transcribed spacer sequences of rDNA,and it was identified as Penicillium oxalicum Currie & Thom.The fragment of the mature phytase was cloned directly from DNA of P.oxalicum RCEF4908 with the special primers designed from phytase gene in GenBank No.AY071824.The de-duced amino acid sequence of this phytase with the conserved motifs RHGXRXP and HD belongs to a typical histidine acid phosphatases.The phytase was overexpressed in the recombinant P49,and its specific activity could reach 8 000 U·mL-1 after induced 144h at optimum pH 5.5,and showed high activity with phytic acid sodium salt at a pH range of 2.5 to 7.0.This recombinant phytase studied here can retain 53.8%,52.5% and 45.2% of its activ-ity after denaturation at 90℃ for10,20 and 30 min,respectively.The broad pH optima and high thermostability of the phytase makes it a better candidate for feed-pelleting applications.

以古尼拟青霉生物量和麦角甾醇含量为指标,对古尼拟青霉的高产麦角甾醇培养条件及麦角甾醇提取工艺进行了优化。正交试验结果表明,高产麦角甾醇的最适液体培养基为蔗糖4%,麸皮2%,KH2PO40.025%和NaCl0.05%;响应面实验结果表明,麦角甾醇最佳提取工艺为:提取溶剂浓度为80%乙醇,液料比为200:1,提取时间为65min。

Glutathione peroxidase, one of the most important components of anti-peroxidation system of the organism, plays an important role in the responses of microorganisms to the environmental stress. In this study, three oxidative stress reagents were performed to determine the stress effect on glutathion peroxidase of Isaria farinosa. Results indicated that the early treatment of oxidative stress under three kinds of conditions all made the GPX activity of bacterium cells decreased. In the later period of cultivation, the GPX activity of bacterium cells under H202 stress had quite small difference compared to the contrast. While it were greatly increased compared to the contrast under the methylene blue and nitrosodisulfonate stress. The Pf340 strain showed diverse biological behavior in response to different oxidative stress.

19 species of Cordyceps and its allies from some nature preserves in Anhui Province, southeastern China are reported. They are Cordyceps phymatospora sp. nov., C. bassiana and its anamorph Beauveria bassiana, C. longissima and its anamorph Hirsutella longissima, C. formosana and its anamorph Hirsutella sp., C. gracilioides, C. filiformis and so on. Specimens examined are deposited in Research Center on Entomogenous Fungi, Anhui Agricultural University (RCEFAAU).

A Beauveria bassiana strain Bb21 was isolated from naturally infected green peach aphid Myzus persicae (Hemiptera: Aphididae). The effects of the strain on M. persicae and its two predaceous natural enemies Chrysoperla carnea (Neuroptera: Chrysopidae) and Harmonia axyridis (Coleoptera: Coccinellidae) were investigated under laboratory conditions. Bb21 had strong pathogenicity to M. persicae, with the LD50 of 97 conidia x mm(-2) (45-191, 95% confidence interval), but was less pathogenic to the second instar nymph of C. carnea, with the LD50 of 1089 conidia x mm(-2). The LD50 for C. carnea was 10.2 times higher than that for M. persicae. The pathogenicity of Bb21 to H. axyridis was very weak, with a low infection rate of 13% even at a high concentration 5 x 10(8) conidia x mL(-1). The Bb21 at low conidia concentration had less effect on the developmental period and fecundity of the two predaceous natural enemies. However, when applied at the high concentration 5 x 10(8) spores x mL(-1), Bb21 shortened the larval stage of H. axyridis averagely by 1.4 d and decreased the adult emergence rate and fecundity by 33% and 14%, respectively, and shortened the larval stage of C. carnea averagely by 0.7 d and decreased the adult emergence rate and fecundity by 24% and 11%, respectively. Since the LD50 for green peach aphid was much lower than that for the two predaceous natural enemies, and had very low effect on the adult emergence rate and fecundity of the two predators at the concentration recommended for field spray, Bb21 could be applied as a biocontrol agent of M. persicae in the integrated management of pernicious organisms.