长寿老人源双歧杆菌优良菌株的筛选

以人结肠腺癌细胞系HT-29细胞为试材,对来源于广西巴马百岁以上长寿老人肠道的24株双歧杆菌进行了体外黏附试验。结果发现,双歧杆菌均具有一定的黏附能力,其中TTF、Z2、TZ5和J-1菌株具有较高的黏附能力。进一步对4株初筛双歧杆菌耐胃酸、胆汁酸和合成B族维生素能力的试验发现,双歧杆菌TTF菌株不仅能合成较高的B1、B2、B6、B12等多种B族维生素,而且在pH3．0的条件下处理120min存活率达93．11％,同时在2％胆盐浓度下处理24h有较好的存活,具有显著的综合优势。     

荷叶离褶伞优良菌株筛选

以棉籽壳、木屑、玉米芯等为主料设计了10个栽培配方,研究不同配方下7株荷叶离褶伞的菌丝生长和出菇情况。结果表明,各菌株菌丝颜色以雪白色为主,在玉米芯为主料的配方上菌丝体生长优于以棉籽壳、木屑为主料的配方,但均无子实体形成;以棉籽壳为主料有利于子实体的形成,特别是棉子壳添加少量木屑、树叶、腐殖土、发酵料和小麦更利于子实体形成,菌株3001的出菇周期最短（108d）,产量最高（214.80g/袋）,各菌株出菇的顺序为3001、1035、1004和1013。因此,栽培料组分对荷叶离褶伞菌丝生长及子实体形成影响较大。     

耐高温蛋白酶菌株的筛选

研究了土壤中耐高温蛋白酶菌株的筛选。采用平板透明圈法从7份土样中筛选到3株产蛋白酶菌株G1、G2、G3,运用菌落和菌体形态进行观察,初步鉴定G1、G2为芽孢杆菌属,G3为假单孢菌属。并对它们分别进行逐级升温驯化培养试验,结果表明：三株菌的最适生长温度均为43℃,其中G2、G3菌耐热性较差,在55℃高温下终止产酶,G1菌经驯化培养后,能耐67℃高温,且酶活力达48％,为拓宽酶制剂的应用提供了理论依据。     

蛹虫草优良菌株的筛选

通过对5株蛹虫草的菌丝形态和生长速率、液体培养生物量和胞外多糖、人工栽培和子实体中活性物质虫草多糖和虫草素的对比研究,筛选出优良的蛹虫草菌株。试验结果表明：蛹虫草6号菌株的菌丝的生长速率比其他菌株快;液体培养生物量和胞外多糖含量明显高于其他菌株;人工栽培的蛹虫草子实体头部大,子囊壳丰富,颜色橘黄,出草整齐均匀,出草率高,子实体中虫草多糖和虫草素的含量均高于其他菌株,表明6号菌株具有较高的经济价值,是值得开发和推广的好品种。     

猕猴桃优良株系筛选鉴定研究

报道猕猴桃的芽变选择,雄株选择,各优良株系的生物学特性．果实生长发育规律,产量,果实营养成分和加工性．采用模糊评分选优法筛选鉴定出桂海４号为较理想的优良株系,江西７９－２和桂海１４号为有发展前途的优良株系。     

产植酸酶菌株的分离筛选研究

研究了产植酸酶菌株分离筛选的方法 ,筛选得到分别适宜液态发酵和固态发酵的酶活力较高的黑曲霉XH和黑曲霉PA两菌株。在液态发酵KH2 PO4 对植酸酶产生有促进作用 ,其最佳用量为 8mg %;在固态发酵KH2 PO4 的添加对植酸酶的产生无明显促进作用。 2菌株所产植酸酶具有相似的酶学性质 ,最适作用温度为 5 5℃ ,当底物 pH为 3～ 5具有较高的酶活力。     

有性杂交选育银耳优良菌株的初步研究

为了解决银耳栽培生产中存在菌株优良性状单一,生产效率较低等问题,采用银科1#和Tr21两个不同品种菌株为亲本菌株,通过单孢子分离选取50个有性孢子单核体、两两配对有性杂交试验,显微观察镜检鉴定,统计杂交成功率为36%、杂交菌株与亲本的拮抗试验选育出7株杂交株,栽培对比农艺性状评价选育出1株(GT3)比亲本菌株生长快,生物转化率较高的菌株。试验结果表明有性杂交育种可作为一种行之有效的筛选银耳优良菌株的方法。     

长寿老人源双歧杆菌优良菌株的筛选*

以人结肠腺癌细胞系HT-29细胞为试材,对来源于广西巴马百岁以上长寿老人肠道的24株双歧杆菌进行了体外黏附试验。结果发现,双歧杆菌均具有一定的黏附能力,其中TTF、Z2、TZ5和J-1菌株具有较高的黏附能力。进一步对4株初筛双歧杆菌耐胃酸、胆汁酸和合成B族维生素能力的试验发现,双歧杆菌TTF菌株不仅能合成较高的B1、B2、B6、B12等多种B族维生素,而且在pH 3.0的条件下处理120m in存活率达93.11%,同时在2%胆盐浓度下处理24h有较好的存活,具有显著的综合优势。     

灵芝深层发酵优良菌株的筛选

灵芝种类多,种间与品种间的不同均可能引起有效成分在产量表达上的差异,为了能筛选适宜深层发酵法生产的灵芝菌株,以8个灵芝菌株为研究对象,比较不同菌株在固体培养基上的菌丝体及液体发酵的菌球萌发菌丝生长速度、液体发酵产物、子实体多糖等指标。结果表明:8个菌株菌丝体生长速度有一定差异,以灵芝G9、红芝早萌发且生长速度快,甜芝、紫芝、灵芝G8、血芝其次,灵芝5760、黑芝最慢。8个菌株深层发酵产物由于生物学特性不同而有差别,同时菌株菌丝体生物量与次生代谢产物多糖之间无必然的联系,甜芝菌丝体生物量最大但自溶速度最快,灵芝5760菌球细小,速度最慢,血芝次生代谢产物多糖最高。菌丝体多糖与子实体多糖比较,各个菌株的菌丝体多糖含量均高于子实体多糖含量。提取多糖工艺以超声波辅助破壁处理后,再以热水浸提法提取多糖的得率明显提高,紫芝、黑芝提高了2倍以上。     

筛选选择性降解玉米秸皮的菌株

目的:筛选能选择性降解玉米秸皮的菌株,用于玉米秸皮生物机械法制浆的研究。方法:采用Bavendamm-PDA平板对玉米种植地土壤内的真菌进行初筛选,之后采用玉米秸皮降解实验对产生棕色变色圈的菌株和实验室现有白腐菌进行复筛选。结果:筛选到4株能在Bavendamm-PDA平板上产生棕色变色圈的菌株;降解实验显示杂色云芝引起了最大31.76%的重量损失,重量损失最小的菌株为粗毛栓菌lg-9;除杂色云芝和白耙齿菌降低了玉米秸皮苯醇抽出物外,其他处理样均增加;碳水化合物测定结果表明白耙齿菌处理样综纤维素的损失最大,且各种菌株所引起的综纤维素损失中,纤维素和聚戊糖损失所占的比例有很大差别。结论:粗毛栓菌lg-9对玉米秸皮木素降解的选择性最强,之后为Sd、黄袍原毛平革菌-14和黄袍原毛平革菌-25。     

茯苓健脾作用活性部位的研究

通过建立饮食失节、过食肥甘所致的小鼠脾虚模型,以体重、饮食量、外部表现特征及脾指数为指标,从茯苓总三萜类、茯苓总多糖及茯苓水煎剂中确定具有健脾作用的有效活性部位。结果表明:茯苓总三萜类对脾虚小鼠有明显的恢复作用,茯苓水煎剂也有一定作用,而茯苓总多糖作用不明显。说明茯苓总三萜类是茯苓治疗过食肥甘所致小鼠脾虚证的活性部位。     

茯苓原生质体制备与再生条件的研究

研究了酶、酶解时间、菌龄、稳渗剂等对茯苓原生质体制备与再生的影响。茯苓原生质体制备的最佳条件为:纤维素酶(1．5%)和蜗牛酶(1．5%)的等量混合酶解系统,酶解时间3h,7d菌龄菌丝,产量可达1．77×10~7个/mL。以甘露醇为稳渗剂,采用CYM再生培养基,酶解时间3h,7d菌龄菌丝,其原生质体再生率最高,为0．164%。这一结果为茯苓通过原生质体技术进行菌种改良提供了重要技术参数。     

茯苓中三萜类和多糖类成分的研究进展

主要对中药茯苓中的三萜类和多糖类成分进行了综述。到目前为止已从茯苓的菌核和菌丝中分离到三萜类物质39个,其中羊毛甾-8-烯型三萜12个,羊毛甾-7,9(11)-二烯型三萜16个,3,4-开环-羊毛甾-7,9(11)-二烯型三萜7个,3,4-开环-羊毛甾-8-烯型三萜2个,三环二萜类1个,齐墩果烷型三萜1个;分离到多糖类物质23个。     

Draft Genome Sequence of the Reference Strain of the Korean Medicinal Mushroom Wolfiporia cocos KMCC03342

Wolfiporia cocos is a wood-decay brown rot fungus belonging to the family Polyporaceae. While the fungus grows, the sclerotium body of the strain, dubbed Bokryeong in Korean, is formed around the roots of conifer trees. The dried sclerotium has been widely used as a key component of many medicinal recipes in East Asia. Wolfiporia cocos strain KMCC03342 is the reference strain registered and maintained by the Korea Seed and Variety Service for commercial uses. Here, we present the first draft genome sequence of W. cocos KMCC03342 using a hybrid assembly technique combining both short- and long-read sequences. The genome has a total length of 55.5 Mb comprised of 343 contigs with N50 of 332 kb and 95.8% BUSCO completeness. The GC ratio was 52.2%. We predicted 14,296 protein-coding gene models based on ab initio gene prediction and evidence-based annotation procedure using RNAseq data. The annotated genome was predicted to have 19 terpene biosynthesis gene clusters, which was the same number as the previously sequenced W. cocos strain MD-104 genome but higher than Chinese W. cocos strains. The genome sequence and the predicted gene clusters allow us to study biosynthetic pathways for the active ingredients of W. cocos.     

16种茯苓菌株液体发酵产胞内多糖与三萜的比较研究

筛选茯苓高产胞内多糖和胞内三萜的优良液体发酵出发菌株。采用PDA富集固体平板培养与液体发酵培养测定菌丝体生长速率;采用液体发酵策略分析16种茯苓菌株产胞内多糖与胞内三萜的潜能。实验结果表明菌株生长于固体培养基与种子培养基的生长速率之间没有关联性;降低一级种子培养基初始pH值到4.0时能有效缓解茯苓菌株培养物褐化现象;AS5.137胞内多糖含量最高,达377.60±0.10 mg/g,而DB菌株显示出最高的胞内多糖产量,达1.01±0.13 g/L;Y1菌株胞内三萜含量最高,达83.89±4.28 mg/g,而Jingzhou28菌株胞内三萜产量最高,达136.63±26.66 mg/L。就生产茯苓胞内多糖与胞内三萜而言,AS5.137与DB菌株适合作为液体发酵产胞内多糖的出发菌株;Y1,Jingzhou28,Z(z)与Xingpinzhong菌株均较适合作为液体发酵产胞内三萜的出发菌株。     

茯苓生物学研究和菌核栽培现状及展望

茯苓Wolfiporia cocos是我国药食两用的传统中药,本文对茯苓的分类地位、交配型、生活史、遗传育种、菌核栽培以及组学相关研究进展进行了总结,对有关专利进行了分析。以研究及生产现状为基础,分析了茯苓在基础生物学方面研究的不足、菌核栽培中存在的相关问题,重点分析了交配型研究、子实体诱导形成和生活史研究等重要方面,并对未来茯苓生物学研究和科学栽培进行了展望。     

Response of Wolfiporia cocos to iron availability: alterations in growth, expression of cellular proteins, Fe3+-reducing activity and Fe3+-chelators production

Aims:  The main objective of this study was to evaluate the behaviour of the brown-rot fungus Wolfiporia cocos under differential iron availability.
Methods and Results:  W. cocos was grown under three differential iron conditions. Growth, catecholate and hydroxamate production, and mycelial and extracellular Fe³⁺-reducing activities were determined. Iron starvation slowed fungal growth and accelerated pH decline. Some mycelial proteins of low molecular weight were repressed under iron restriction, whereas others of high molecular weight showed positive iron regulation. Mycelial ferrireductase activity decreased as culture aged, while Fe³⁺-reducing activity of low molecular reductants constantly increased. Hydroxamates production suffered only limited iron repression, whereas catecholates production showed to be more iron repressible.
Conclusions:  W. cocos seems to possess more than one type of iron acquisition mechanism; one involving secretion of organic acids and ferrireductases and/or extracellular reductants, and another relying on secretion of catecholates and hydroxamates chelators.
Significance and Impact of the Study:  This paper is the first to report the kinetic study of brown-rot fungus grown under differential iron availability, and the information provided here contributes to address more traditional problems in protecting wood from brown decay, and also makes a contribution in the general area of the physiology of brown-rot fungi.     

稻曲病两个白化菌株的分离与生物学特性

为了筛选带有自然标记的稻曲病菌菌株,2010年从浙江省象山县和陕西省勉县采集和分离到2个稻曲病白化菌株,ZJa0201和SXa0101。它们在PSA培养基上的生长速度约为其他稻曲病菌株的3倍,未见产生厚垣孢子;在PS培养基上只能产生少量分生孢子。rDNA-ITS和rDNA-IGS序列分析表明,两个白化菌株也与稻曲病菌已知所有菌株的ITS序列同源性高于99.6%;rDNA-IGS序列也属于最为常见的类型,含有2个77bp的重复单元序列。由此推断,这两个白化菌株属于稻曲病菌产孢退化的突变体。白化菌株在PSA上     

1H NMR based metabolite profiling for optimizing the ethanol extraction of Wolfiporia cocos

Metabolite profiling of Wolfiporia cocos (family: Polyporaceae) had been much advancement in recent days, and its analysis by nuclear magnetic resonance (NMR) spectroscopy has become well established. However, the highly important trait of W. cocos still needs advanced protocols despite some standardization. Partial least squares discriminant analysis (PLS-DA) was used as the multivariate statistical analysis of the ¹H NMR data set. The PLS-DA model was validated, and the key metabolites contributing to the separation in the score plots of different ethanol W. cocos extract. ¹H NMR spectroscopy of W. cocos identified 33 chemically diverse metabolites in D₂O, consisting of 13 amino acids, 11 organic acids 2 sugars, 3 sugar alcohols, 1 nucleoside, and 3 others. Among these metabolites, the levels of tyrosine, proline, methionine, sarcosine, choline, acetoacetate, citrate, 4-aminobutyrate, aspartate, maltose, malate, lysine, xylitol, lactate threonine, leucine, valine, isoleucine, uridine, guanidoacetate, arabitol, mannitol, glucose, and betaine were increased in the 95% ethanol extraction sample compared with the levels in other samples, whereas level of acetate, phenylalanine, alanine, succinate, and fumarate were significantly increased in the 0% ethanol extraction sample. A biological triterpenoid, namely pachymic acid, was detected from different ethanol P. cocos extract using ¹H-NMR spectra were found in CDCl₃. This is the first report to perform the metabolomics profiling of different ethanol W. cocos extract. These researches suggest that W. cocos can be used to obtain substantial amounts of bioactive ingredients for use as potential pharmacological and nutraceuticals agents.