枣树越冬期内生真菌的分离鉴定及抑菌研究

为了明确越冬期枣树组织中内生真菌的种类和分布情况,并从中筛选出对枣缩果病病原有拮抗作用的内生菌株,从越冬期健康枣树的树皮、枣股、枝条中分离获得内生真菌225株,共鉴定出12个真菌属,其优势属为茎点霉属、链格孢属、毛壳菌属和镰刀菌属。研究结果表明:越冬期健康枣树组织中内生真菌分布普遍,种类丰富;不同品种、不同部位中内生真菌的种类和数量都有差异。室内平板对峙实验结果表明:在225株内生真菌中,有25株内生真菌对枣缩果病病原茎点霉菌1-2有抑制作用,最高抑菌率为85.45%;有14株内生真菌对病原壳梭孢菌1-5有抑制效果,最高抑菌率为76.00%;内生真菌F7和F8对两种病原菌均有抑菌效果,而且均有明显的抑菌带,这说明上述两株菌均可产生抗菌代谢物质。     

Pyrenocines J-M: four new pyrenocines from the endophytic fungus, Phomopsis sp

Four new pyrenocines, named pyrenocines J-M (1-4), were isolated from an endophytic fungus, Phomopsis sp, by a bioassay-guided method. The structures of the new compound have been assigned from 1H and 13C NMR spectra, DEPT, and by 2D COSY, HMQC, and HMBC experiments. Preliminary studied showed that compounds 1, 2 and 4 showed good antifungal, antibacterial, and algicidal properties, while compound 3 had good antibacterial and algicidal properties.     

东北红豆杉内生真菌的鉴定-中国新记录属种及其代谢产物

从长白山自然保护区原始森林200多年生东北红豆杉(TasxuscuspidataSiebetZucc.)树皮中分离94株内生真菌,鉴定出19种。其中,除2个新种、1个新变种和6个国内已知种(已详细描述并发表在《林业研究》2003,14(4):290-294),本文还对包括9个新记录属种的形态特征进行了详细描述:坚孢葡萄单孢(StaphylotrichumcoccosporumMeyeretNicot)、砖红葡萄痕孢(BotryodemalateritiumPapendoorf&Upadhyay)、球头孢[Oedocephalumglomerulosum(RulliardSacc.)、球头三型孢(TrichosporonoidesoedocephalisHaskins&Spencer)、扁圆球孢[Beniowskiasphaeroidea(KalChbrenneretCaeke)Mason]、疣顶孢(ThermomycesverrucosusPugh,Blakeman&Morgan-jones)、腐殖厚壁孔孢(GilmaniellahumicolaBrron)、球乳突孢[Papulariasphaerosperma(Pers.:Fr.)vonHohnel]、褐钝孢[Ambrosiellabrunnea(Verrall)Batra]和1个新记录种:假线壳囊孢(CytosporinanothaDied.)。对中国新记录属种内生真菌,提供了形态学特征描述与图解及其生境、寄主的记载。用薄层层析技术,测定了各菌种发酵代谢产物可与紫杉烷类物质显色剂棗香草试液和稀碘化铋钾试液不发生明显的颜色反应。图10参23。     

灰背杜鹃菌根真菌的分离与鉴定及DGGE的应用

应用传统的培养方法分离菌根真菌,应用ITS rDNA分子生物学手段,进行了菌株的分类鉴定,并结合DGGE(变性梯度凝胶电泳)技术对菌根真菌进行检测。传统培养得到了2种与杜鹃花菌根真菌亲缘关系十分密切的真菌,分别为Phialocephala fortinii和Epacris pulchella root associated fungus。DGGE(变性梯度凝胶电泳)技术检测到1种菌根真菌为Rhizoscyphus ericae。     

油茶总RNA及mRNA的分离与纯化

利用改良的CTAB法,从富含酚类糖类等次生物质油茶叶片中分离出总RNA,并从总RNA中,通过两次过Oligo(dT)柱分离纯化得到mRNA.实验结果证明:所提取的RNA和mRNA完整,质量较高,并应用到油茶cDNA文库的构建.     

Identification of Taxus cuspidata Sieb. et Zucc. endophytic fungi-new species, species known and their metabolite

A total of 94 isolates of endophytic fungi were isolated from the bark of 200-yr.-old Taxus cuspidata Sieb. et Zucc.in the primeval forest of the Changbai Mountain Natural Reserve, and 19 species of endophytic fungi were identified, including 10 new recorded-genus-species, 2 new species (Phomopsis Iongiscoleosporu Y. Xiang et Lu An Guo Wu Wen Fang, Coniothyrium macrospoum Y. Xiang J.X. et Lu An Guo Wu Wen Fang), 1 new varied species (Alternaria alternata (Fr.) Keissler var. taxi Y.Xiang et Lu An Guo) and 6 known species of China (Eurotium amstelodomi Mgngin, Eurotium repens de Bary, Botrytis sp.,Penicillium citrinum Thom, Epicoccum nigrium LinK, Fusarium sp.). Through thin layer chromatography (TLC), liquid fermentation metabolite of the strains was determined, and four strains (Alternaria alternata (Fr.) Keissler var. taxi Y. Xiang et Lu An Guo Wu Wen Fang, Botrytis sp., Eurotium amsteloodomi Mgngin, Eurotium repens de Bary) were screened out, whose metabolites reacted positively with the vanillic aldehyde that was one special taxoid developer. Among the four strains, Alternaria alternata (Fr.) Keissler var. taxi Y. Xiang et Lu An Guo, produced one compound largely, which positively reacted with one alkaloids developer-Bismuth potassium iodide. The compound is identified as taxoids type through spectrum analysis. This demonstrates that Alternaria alternata (Fr.) Keissler var. taxiY. Xiang et Lu An Guo can highly produce taxoids largely.     

高速逆流色谱法分离制备太白贝母中贝母辛

运用高速逆流色谱(HSCCC)技术从太白贝母中分离制备贝母辛。以正己烷/乙酸乙酯/甲醇/水(体积比4∶5∶3∶5)为溶剂体系,转速800 r/min,流速2.0 m L/min,温度35℃,检测波长为215 nm,从50 mg太白贝母总生物碱中经一次HSCCC分离可得到5.3 mg贝母辛,纯度为96.3%,回收率为87.2%。并利用高效液相色谱-四极杆飞行时间质谱(HPLC-QTOF MS)对贝母辛进行了结构确证。     

三羟鹅膏毒肽的反相高效液相色谱分离纯化

用反相高效液相色谱法,以0.02mol醋酸铵-乙腈为流动相和梯度洗脱模式,从灰花纹鹅膏菌中分离纯化出三羟鹅膏毒肽(amanin),产品纯度达96%以上,三羟鹅膏毒肽的分子量为903.3.     

云南松纵坑切梢小蠹寄生真菌种类调查及其应用技术研究

在云南主要蠹害林区，采集松纵坑切梢小蠹异常虫体７６号，在室内进行分离培养、纯化鉴定，致病测定后，到林间接种试验。筛选出对松纵坑切梢小蠹致病性强的本色镰刀菌ＦｕｓａｒｉｕｍｃｏｎｃｏｌｒＲｅｉｎｒ，球孢白僵菌Ｂｅａｕｖｅｒｉａｂａｓｉａｎａ（Ｂａｓｌｓ）Ｖｕｉｌ．粘孢白僵菌ＢｅａｕｖｅｒｉａｖｅｌａｔａＳａｍｓｏｎｅｔＥｖａｎｓ，青霉菌Ｐｅｎｉｃｉｌｉｕｍｓｐ．拟青霉菌Ｐａｅｃｉｌｏｍｙｃｅｓｓｐ．等５种寄生真菌。经林间蛀干实验，小蠹带菌在干内死亡率分别为８５．６％，８２．１％，８０．５％，８１．５％，８２．１％，９０．１％。     

Susceptibility of longleaf pine roots to infection and damage by four root-inhabiting ophiostomatoid fungi

Restoration of longleaf pine-dominated uplands is common on many public and private lands throughout the southeastern United States. The once dominant longleaf pine ecosystem is important to many now-threatened and endangered plant and animal species, and land managers are increasing efforts to reestablish this fire-dependent forest. Unfortunately, tree mortality in longleaf pine has been observed following attempts to re-introduce prescribed fire. Root-inhabiting ophiostomatoid fungi and their insect vectors have invaded roots of symptomatic longleaf pine. Although, the relationship between ophiostomatoid fungi and longleaf pine roots is poorly understood. In order to assess the pathogenicity and virulence of four ophiostomatoid fungi to longleaf pine, trees within two broad age classes (20–30 and 40–60 years) were used for root inoculations during the fall of 2006 and 2007 along with the spring of 2007 and 2008. All fungal species consistently caused resin-filled, discolored lesions on the phloem surface extending to the xylem. The successful inoculation of healthy longleaf pine roots confirms the pathogenicity of Grosmannia huntii, Leptographium procerum, Leptographium serpens, and Leptographium terebrantis. G. huntii caused the largest lesions, including 22.20 cm², 13.37 cm², and 9.21 cm² larger than L. procerum, L. terebrantis, and L. serpens respectively. In contrast, L. procerum caused significantly smaller lesions than all other fungi, including 8.65 cm² smaller than L. terebrantis and 10.69 cm² smaller than L. serpens. Restoration efforts of longleaf pine may be affected by fungal root infection in the future. Future studies should focus on the interactions between stress factors associated with longleaf pine to define more clearly the ecological role of root-inhabiting ophiostomatoid fungi in the ecosystem.     

红绒盖牛肝菌发酵培养基筛选及液体培养条件

在摇瓶液体培养条件下,研究了14种不同成分培养基对红绒盖牛肝菌菌丝体生长的影响及液体培养条件,结果表明,红绒盖牛肝菌最适发酵培养基为:蔗糖2%、玉米粉2%、黄豆粉1.5%、酵母膏0.5%、MgSO40.05%、KH2PO40.1%。培养基中添加有机硅类消泡剂对红绒盖牛肝菌菌丝体生长没有影响;液体发酵的适宜初始pH为5～6;液体培养周期为10 d,菌丝体生物量可达5.7 g/L。     

黄芪鲨烯合酶基因的克隆和序列分析

参照豆科其他植物的鲨烯合酶基因序列设计了引物,通过逆转录PCR方法,从荚膜黄芪（Astragalus mem-branaceus（fish.）Bge）中克隆了鲨烯合酶基因cDNA序列（GenBank登录号HQ829974）。通过BLAST发现黄芪鲨烯合酶推测的氨基酸序列与大豆鲨烯合酶（BAA22559.1）序列相似性最高,达到了88%。黄芪鲨烯合酶的克隆为以后通过基因转化提高黄芪皂苷含量亦或通过生物工程手段生产黄芪皂苷打下基础。     

竹笋胰蛋白酶抑制剂的分离纯化及其性质

为寻找天然胰蛋白酶抑制剂,经过葡聚糖凝胶G25、G50柱、二乙二胺乙基纤维素－葡聚糖凝胶A50分离纯化从竹笋中获得了竹笋胰蛋白酶抑制剂(BSTI),纯化后浓度比粗提物提高了1200倍,SDS-PAGE显示纯化胰蛋白酶抑制剂为单一条带。其性质研究表明：经100℃热变性15 min后,BSTI仍保持50%的抑制活性,表明BSTI有较强的热稳定性;BSTI的等电点为4.4左右; BSTI在CoCl2溶液中还有94.7%的活性,但是在FeSO4溶液中活性却完全被压制;BSTI在氧化剂中活性不高;几种还原剂对其活性有一定的抑制,但是维生素C不仅不会抑制其活性反而能够提高它的活力;螯合剂KCN能大幅提高BSTI的活性,EDTA和喔星会抑制其活性。     

银杏叶中黄酮和萜类内酯分离工艺条件优化

用闪式硅胶柱层析分离纯化银杏叶中有效成分黄酮醇苷和萜类内酯工艺条件的优选。用不同极性溶剂(乙醚、乙酸乙酯、丙酮、乙醇)对加样品后的闪式硅胶层析柱进行洗脱,收集洗脱液,采用高效液相色谱法分别测定每份洗脱液中黄酮醇苷和萜类内酯的含量,从而筛选出应用闪式硅胶柱层析时,最佳的洗脱液种类和用量为:萜类内酯用3倍床体积的乙酸乙酯洗脱,黄酮醇苷用4倍床体积的丙酮洗脱即得。     

绵马贯众多酚的分离纯化及其抗氧化活性研究

采用70%乙醇提取绵马贯众多酚,并通过吸附实验考察了7种大孔树脂对绵马贯众多酚粗提物吸附率和解吸率的影响,发现HPD100树脂对绵马贯众多酚的纯化效果较好;进一步分析了样品质量浓度、上样速度、解吸乙醇体积分数、洗脱速度及样品pH值对HPD100树脂纯化绵马贯众多酚的影响,得出最佳纯化工艺条件为:样品溶液质量浓度为2.00 g/L,pH值为5,上样速度为1 mL/min,解吸乙醇体积分数为70%及洗脱速度为2 mL/min。该工艺条件下,绵马贯众多酚纯度由纯化前的27.64%提高到纯化后的54.00%(得率为6.46%),纯化效果显著。纯化后绵马贯众多酚抗氧化活性显著增加,对DPPH自由基(DPPH·)及羟基自由基(·OH)的IC₅₀值分别为0.01和0.49 g/L(纯化前为0.05和1.79 g/L),0.04 g/L时对DPPH·的清除率为90.82%,与Vc相当;质量浓度为0.05 g/L时,纯化后绵马贯众多酚对铁离子还原能力为0.47,明显高于纯化前的0.25;且在酸性条件下DPPH·清除能力以及铁离子还原能力更高,在碱性条件下·OH清除能力更高。     

杜仲总木脂素及松脂醇二葡萄糖苷的纯化研究

通过大孔吸附树脂-C18反相硅胶柱层析-正相硅胶柱层析联用技术,对杜仲总木脂素及松脂醇二葡萄糖苷(PDG)进行纯化研究。结果表明,含总木脂素23.33%和松脂醇二葡萄糖苷粗品0.960 5%的杜仲提取液经AB-8树脂一次纯化,可以制得71.61%的杜仲总木脂素有效部位以及8.070%的松脂醇二葡萄糖苷粗品,再经C18反相硅胶柱层析纯化,得到53.12%的松脂醇二葡萄糖苷。最后通过正相硅胶柱层析进一步纯化,得到91.50%的松脂醇二葡萄糖苷产品。     

水竹叶总黄酮的提取及纯化工艺研究

以水竹叶为原料，采用水浴和超声波两种提取方法，通过单因素与正交试验确定了水竹叶总黄酮提取的最佳条件，并筛选了不同树脂对总黄酮的吸附和解吸附特性。实验结果表明：水浴提取最佳条件为：温度50℃，提取时间2．0h，乙醇体积分数60％，固液比1：20（g：mL，下同）；总黄酮得率1.65％。影响得率的因素为：固液比〉乙醇体积分数〉提取温度〉提取时间。超声波提取最佳条件为：2．0g水竹叶，超声波作用时间30min，乙醇体积分数60％，固液比1：15；总黄酮得率为1．27％。影响得率的因素为：超声波作用时间〉乙醇体积分数〉固液比。AB-8树脂对总黄酮纯化具有较好效果，体积分数为70％的乙醇是较好的解吸剂。     

沙棘果废渣中原花青素提取纯化工艺研究

以儿茶素为对照,沙棘原花青素得率为考察指标,通过正交试验法优选提取沙棘果废渣中原花青素的最佳工艺,并在此基础上,采用大孔树脂吸附、分离、纯化原花青素提取物。结果表明,超临界CO2脱脂后的沙棘果废渣中原花青素最佳提取工艺参数为：70%（体积分数）乙醇作溶剂,料液比1∶7,提取温度40℃,提取时间1 h;D-101型树脂对沙棘果废渣中原花青素吸附能力较强,纯化后的原花青素纯度可达73.3%.     

金银花中绿原酸的提取及纯化方法比较

主要介绍了近10年来金银花中绿原酸的提取及纯化工艺,重点对各种方法进行了分析、论述和比较,并且结合实际生产应用的可行性,提出了合理的操作工艺:先用水提法提取出金银花中的有效成分,再用大孔树脂法对提出物进行纯化,最终可得到提取率及纯度都较高的绿原酸。