培养基及培养条件对冬虫夏草菌固体发酵产分生孢子的影响

为获得冬虫夏草菌固体发酵产分生孢子的最优工艺,以野生分离的冬虫夏草菌为材料,对其固体发酵产分生孢子的培养基及培养条件进行了研究。试验结果表明：泥炭土为最佳基础培养基,该培养基中冬虫夏草菌气生菌丝生长一般,但产分生孢子最多,可达4.2×10³个/g;泥炭土培养基中添加0.1‰ IAA（吲哚乙酸）、0.1‰ IBA（吲哚丁酸）和0.1‰ NAA（萘乙酸）能促进冬虫夏草菌气生菌丝的生长和分生孢子的产生,其分生孢子达8.1×10³个/g;该基础培养基中,冬虫夏草菌于18℃培养30d后,在10℃、相对湿度45%、蓝光照射进行诱导,分生孢子可达1.0×10⁴个/g。本研究建立了一种大量获取冬虫夏草菌分生孢子的方法,为冬虫夏草繁育奠定了基础。     

冬虫夏草原生质体形成和再生的初步研究

研究冬虫夏草菌丝体的菌龄、酶种类、酶解温度、酶解时间、pH、稳渗剂和几种再生培养基对原生质体形成和再生的影响。最佳条件为 :生长 6d的菌丝体 ,组合酶 (1%蜗牛酶 +1%纤维素酶 ) ,酶解温度 36℃ ,酶解时间 2 .5h,pH6 .4 ,稳渗剂 0 .4M甘露醇溶液 ,RM3再生培养基。在此条件下原生质体的形成为 2 .0 4× 10 9个 ml,再生率为 0 .0 91%。     

榅桲的组织培养和快速繁殖

植物名称:榅桲(Cydonia oblonga)。材料类别:早春将榅桲的一年生枝条,在温室中水培10—15天,待其开始萌动时,剪成带1—2个腋芽的茎段,经常规消毒后,在无菌条件下剥离茎尖培养。培养条件:茎尖生长培养基:MS+BA0.3mg/L(单位下同)+GA 0.5+NAA 0.2。增殖培养基:MS+BA 1+IAA 0.2。生根培养基:1/2MS+     

不同培养基对冬虫夏草菌丝生长的影响

不同培养基对3个来源不同的冬虫夏草菌菌丝生长影响的研究结果表明,在不同培养基上,各菌株之间在菌丝萌发时间、40d菌落直径和菌落形态上表现出一定的差异。同时筛选到的最优培养基比已报道培养基的菌落生长速度快2～3倍,且菌丝为白色、粗壮,菌丝质量较优。     

冬虫夏草菌丝生长的营养需求*

从青海玉树洲巴塘山采集的野生冬虫夏草中分离到一株冬虫夏草菌,通过该菌对碳源、氮源、无机盐和维生素的利用试验,筛出该菌菌丝液体培养的最优条件:葡萄糖1.5%、麦芽汁1%、蛋白胨1%、酵母膏0.5%、pH6.0,温度23℃,接种量5—10%,摇床转速80—90r/min。     

野生冬虫夏草与蝙蝠蛾被孢霉发酵菌粉主要营养成分分析与比较

目的:分析与比较蝙蝠蛾被孢霉发酵菌粉与野生冬虫夏草的主要营养成分。方法:采用干燥法测定的水分含量,用恒量法测定灰分含量,用酸水解法测定粗脂肪含量,用凯氏定氮法测定粗蛋白含量,用重量法检测粗纤维和粗多糖含量,采用HPLC检测氨基酸和核苷类物质含量,采用GC测定脂肪酸含量,采用原子吸收法测定微量元素种类及含量。结果:发酵菌粉与野生冬虫夏草均属于高蛋白、高碳水化合物、低脂肪的食品原料,而菌粉中低含量的粗纤维更利于人体对食物的消化和营养吸收。菌粉含有比野生冬虫夏草更丰富的氨基酸(19种)和多不饱和脂肪酸种类(13种)。同时,微量元素的种类和比例也更加合理,有害元素As的含量显著低于野生冬虫夏草。结论:蝙蝠蛾被孢霉发酵菌粉的营养价值可以与野生冬虫夏草相媲美,具有良好的开发应用前景。     

丁香杂交胚的组织培养

植物名称:丁香属植物(Syringa)。材料类别:①授粉40～50天的丁香(Syringameyeri×S.microphylla)杂交胚;②授粉60～110天的丁香。(Syringa persica×S.VuZgarls"Albaplena”)杂交胚。培养条件:基本培养基为MS培养基,蔗糖浓度3％,pH=5.8,培养温度25±2℃,光照强度1000～2500lx,每日光照14小时。杂交组合①取授粉后40～50天的杂交幼胚进行培养。杂交组合②取授粉后60～110天的杂交胚培养,从授粉后60天开始,每周取胚培养1次。当授粉90天后,每隔1天取胚培养1次,直到110天     

桔梗叶片的离体培养和植株再生

植物名称:桔梗(Platycodon grandiflorum)。材料类别:叶片。选两年生桔梗植株,在花蕾形成前后分批采摘叶片,均用饱和漂白精片水溶液消毒20分钟,无菌水冲洗2—3次,分别接种。培养条件。以MS为基本培养基,添加水解乳蛋白500mg/l,将 NAA0.5ppm和 6-BA1ppm配比组成培养基Ⅰ,MS+2,4-D 0.2ppm组成培养基Ⅱ,培养基Ⅲ为1/2 MS+IBA 0.5mg/l。材料接种后,先在黑暗或微光下培养25—30天,再移到光照度2000lx的条件下培养,培养温度为20—28℃。     

局限青霉生淀粉水解酶的产生条件及酶一般性质的研究

生淀粉水解酶产生菌局限青霉(Penicillium restrictum)127-2是由土壤中分解得到的,具有较强的降解生淀粉能力,水解1g生淀粉需8.0国际单位酶量.Sasaki等曾报道草酸青霉1-9、变幻青霉的培养液能分解生淀粉,但未报道酶的形成条件;Takao等研究了罗尔伏革菌生淀粉酶的形成条件和酶的一般性质.本文介绍局限青霉127-2生淀粉水解酶的形成条件和酶的一般性质.1 材料和方法1.1 菌种培养1.1.1 斜面培养:用查氏琼脂斜面培养基在28—30℃培养5—7天.1.1.2 三角瓶振荡培养:于250ml三角瓶中加40ml发酵培养基,经1kg/cm~2 30分钟灭菌,冷却后接种,置于28—30℃的摇床(200r/min)振荡培养4—5天.1.2 酶活力测定     

糖类对冬虫夏草菌Ophiocordyceps sinensis芽生孢子产量及毒力的影响

冬虫夏草Chinese cordyceps是食药两用的传统名贵资源。其人工培养需要冬虫夏草菌Ophiocordyceps sinensis的优质芽生孢子。为获得高产量及高感染力芽生孢子,本研究测定不同糖类培养基对冬虫夏草菌芽生孢子产量及毒力（对小金蝠蛾Thitarodes xiaojinensis幼虫的存活率、携菌率和僵虫率）的影响。两个冬虫夏草菌株于含6种糖（葡萄糖、果糖、阿拉伯糖、麦芽糖、蔗糖和海藻糖）的液体培养基中分别培养30、45和60 d,计数芽生孢子产量并将收集的孢子注射感染两个品系的小金蝠蛾6龄幼虫,置于不同温度下观察幼虫的存活率、携菌率及僵虫率。结果显示：不同糖源培养基中芽生孢子的产量差异显著,其中含麦芽糖培养基中芽生孢子产量最高;注射菌株和幼虫品系对被注射幼虫的存活率无显著差异;不同糖源培养基获得的芽生孢子对蝠蛾幼虫的存活率无显著差异,对携菌率和僵虫率产生影响;以含麦芽糖为糖源的培养基、芽生孢子培养时间显著影响被注射幼虫的存活率与僵虫率,其中培养30 d的冬虫夏草菌所注射的幼虫的存活率、僵虫率显著高于60 d的;温度也显著影响被注射幼虫的存活率、僵虫率,10℃的幼虫存活率和僵虫率显著高于14℃、18℃的。研究结果为培养优质冬虫夏草芽生孢子,提高被侵染蝙蝠蛾幼虫僵虫率提供了参考。     

SOME FACTORS AFFECTING GROWTH OF APHANOMYCES EUTEICHES IN SYNTHETIC MEDIA

Papavizas , G. C., and C. B. Davey . (USDA, ARS, Crops Research Division, Beltsville, Maryland.) Some factors affecting growth of Aphanomyces euteiches in synthetic media. Amer. Jour. Bot. 47(9) : 758–765. Illus. 1960.—Some nutritional requirements of 3 single-zoospore isolates of Aphanomyces euteiches Drechsler were studied in a synthetic medium (SM-1) consisting of mineral salts, D-glucose, DL-glutamic acid, and thioglycolic acid. Micronutrients were essential for growth, whereas vitamins were not. Controlled pH experiments showed that the fungus has a relatively wide pH range (5.2–7.7) for growth with an optimal range between 5.4 and 6.5. When the pH of the medium was maintained within the optimal range by periodic adjustments, ammonium nitrogen was utilized as the sole source of nitrogen. Nitrate nitrogen was unavailable to the fungus under all conditions. DL-glutamic acid was superior as a nitrogen source to either the D or the L isomer. Growth rate and final mycelial dry weight in a synthetic medium (SM-2) containing 14 amino acids in proportions found in 1.0% solution of yeast extract approximated those obtained in complex media. Mycelial dry weights in SM-2 were superior to those obtained in the glucose-glutamic acid-thioglycolic acid medium, but this superiority was due to DL-methionine, a sulfur-containing compound of the amino acid mixture. Sulfates were not utilized as sole source of sulfur, whereas cysteine, cystine, and thioglycolic acid supported fair amounts of growth. The optimal sulfur concentrations from thioglycolic acid for growth in SM-1 were between 128 and 192 mg/1. Mycelial yields equalling or exceeding those supported by complex media were obtained with high concentrations of D-glucose and DL-glutamic acid.     

Improvement of insulin resistance and insulin secretion by water extracts of Cordyceps militaris, Phellinus linteus, and Paecilomyces tenuipes in 90% pancreatectomized rats

The effect of supplementation with Phellinus linteus (P. linteus), Paecilomyces tenuipes (P. tenuipes), and Cordyceps militaris (C. militaris) mushroom water extracts on the insulin secretion and insulin resistance of 90% pancreatectomized (Px) male Sprague Dawley rats was investigated. Px rats were daily administered 0.5 g of P. linteus, P. tenuipes, and C. militaris aqueous extracts or a placebo per 1 kg body weight with a 40% fat diet for 8 weeks. Fasting serum glucose levels were lower in rats receiving C. militaris than in the control group. Insulin secretion at the elevated serum glucose levels was lowest in rats that consumed P. tenuipes in hyperglycemic clamp. Whole body glucose disposal rates increased in C. militaris but decreased in P. tenuipes compared to those in the control group in euglycemic hyperinsulinemic clamp. The GLUT4 content and fraction velocity of glycogen synthase in the soleus and quadriceps muscles increased in the rats treated with C. militaris, but P. tenuipes decreased both. In sum, a water extract of C. militaris ameliorates insulin resistance by enhancing glucose utilization in skeletal muscles.     

大别山地区虫生真菌群落结构与生态分布

本文对安徽大别山地区虫生真菌的群落结构和生态分布规律进行了研究。在天堂寨和鹞落坪2个自然保护区共设样方278个,先后采集罹病标本1986个,共计50种,隶属于4目、4科、16属。优势种依次为粉拟青霉、细脚拟青霉、下垂虫草、球孢白僵菌和金龟子绿僵菌。在垂直分布上,种的丰度、多度和物种多样性指数都以海拔1100-1150m最高,随着海拔升高或降低,各多样性指标基本上表现递减的趋势。季节分布上,夏季是虫生真菌物种分布最丰富的季节,随着温度下降和雨量的减少,种的丰度和多度也明显下降。另外,该区虫生真菌群落呈现出明显的水平地带性分布规律。     

细脚拟青霉培养性状和药理作用的初步研究

细脚拟青霉被认为是蛾蛹草(Cordyceps polyarthra Moller)的不完全阶段。在自然基物上生长最佳,容易培养。该菌生长最适温度为24—26℃,40℃分生孢子失活。无性世代周期观察表明,从孢子萌发到次代分生孢子产生仅24小时。药理试验结果,该菌对小鼠毒性口服剂量LD_(50)>70g/kg;腹腔注射 LD_(50)为17.6±1.78g/kg。并对小鼠常压耐缺氧能力、镇静、镇痛等作用进行了实验,和对照相比均有非常显著的差异。且毒性较低,可能成为一种有药用价值的资源菌。     

Improvement of lipid production from an oil-producing filamentous fungus, <Emphasis Type="Italic">Penicillium brevicompactum</Emphasis> NRC 829, through central composite statistical design

In the present study, 13 filamentous fungi were screened for their lipid production and an oleaginous fungus, Penicillium brevicompactum NRC 829, was found to be the highest lipid producer. Screening of various agro-industrial residues was performed and sunflower oil cake proved to be the best substrate for lipid production. A central composite design was employed to investigate the optimum concentrations of the most significant medium components required to improve the lipid production by P. brevicompactum. The results clearly revealed that the maximal lipid production of 8.014 ± 0.06 gL^?1 (representing 57.6% lipid/dry biomass) was achieved by the fungus when grown for 6 days at 30 °C under static condition in a medium containing sunflower oil cake, NaNO₃ and KCl at final concentrations of 8, 0.75 and 0.25 gL^?1, respectively. Gas chromatography-mass spectrometry analysis of P. brevicompactum lipid indicated that linoleic acid (LA) (C18:2–6, 9) was the most abundant fatty acid, accounting for up to 62% of the total fatty acid profile, followed by palmitoleic acid (C16:1, 16%) and linolenic acid (C18:3, 8%). These results suggest that P. brevicompactum NRC 829 may have potential for commercial development for the production of LA by fermentation using cheap raw material.     

Nutritional requirements and media development for Lactococcus lactis IL1403

Lactic acid bacteria are extensively used in food technology and for the production of various compounds, but they are fastidious in nutrient requirements. In order to elucidate the role of each component precisely, defined multicomponent media are required. This study focuses on determining nutrient auxotrophies and minimizing media components (amino acids, vitamins, metal ions, buffers and additional compounds) for the cultivation of Lactococcus lactis subsp. lactis IL1403, using microtitre plates and test tubes. It was shown that glutamine and asparagine were the most important media components for achieving higher biomass yields while the branched-chain amino acids were necessary to increase specific growth rate. The amino acid and glucose ratio was reduced to achieve minimal residual concentration of amino acids in the medium after the growth of cells, whereas the specific growth rate and biomass yield of cells were not considerably affected. As the percentage of each consumed amino acid compared to initial amount is larger than measurement error, these optimized media are important for achieving more precise data about amino acid utilization and metabolism.     

The Influence of Calcium or Manganese on the Resistance to EDTA, Polymyxin B or Cold Shock, and the Composition of Pseudomonas aeruginosa Grown in Glucose- or Magnesium-depleted Batch Cultures

Cultures were batch grown in simple salts media in which growth was limited either by depletion of glucose and magnesium (C/Mg-dep) or by glucose alone (C-dep). Cultures were also grown in these media supplemented by calcium and/or manganese.
All cultures grown in the C-dep media were sensitive to ethylenediaminetetraacetic acid (EDTA), polymyxin and also to cold shock but were relatively resistant to ethyleneglycol-bis(2-aminoethyl ether)-N, N-tetraacetic acid (EGTA). Inclusion of calcium or manganese in the growth medium enhanced lysis by EDTA. Cultures grown in the basic C/Mg-dep media were resistant to EDTA, EGTA, polymyxin and to cold shock. Sensitivity to these agents was retained by cultures grown in C/Mg-dep media supplemented with Ca²⁺ and/or Mn²⁺. Cells grown in C/Mg-dep media with added Mn²⁺ were more sensitive to EDTA and polymyxin than those from the unsupplemented C/Mg-dep media but still resistant compared with C-dep cultures. All cultures from supplemented C/Mg-dep media were more sensitive to EGTA than those from any of the C-dep media.
Whole cells and cell walls from these various media had differing amounts of cell wall, phosphorus, amino sugar, carbohydrates, readily extractable lipid (REL), total phospholipid (PL), and especially differences in cell wall divalent metal cation content.
The differences in PL, REL and amino sugars and carbohydrate did not correlate with the response of C-dep and C/Mg-dep bacteria to EDTA, EGTA or polymyxin. The results are discussed in relation to the hypothesis that the sensitivity of Pseudomonas aeruginosa to polymyxin and EDTA is more dependent on outer membrane cation content rather than on other components, e.g. PL and lipopolysaccharide.     

淡水育珠蚌体外培养外套膜细胞分泌的珍珠质的性质研究

对淡水育珠蚌分泌珍珠质的外表皮进行组织培养,取培养不同时间的培养基,用氨基酸分析仪对其氨基酸测定的结果与空白培养基作比较,其中珍珠所含的各种氨基酸均增加,尤其是珍珠中含量最高的丙氨酸,甘氨酸和谷氨酸增加最多,珍珠药用有效成分之一的牛磺酸,随组织培养时间的延长而逐渐增加,用等离子光谱测定培养组织匀浆液与未培养比较,钙的含量也大为增加,结果表明,离体组织培养分泌的珍珠质的化学成分和性质与活体基本相同。     

六种彩叶植物营养成分及有害元素含量分析

以六种彩叶植物紫叶李、红花木、红叶石楠、红枫、鸡爪槭和金叶女贞的叶片为材料,对其营养成分及有害元素含量进行了测定及分析。结果表明:蛋白质和可溶性糖含量以红花木为最高,维生素C含量以红枫为最高,β-胡萝卜素含量差异最大,紫叶李最高达15.99mg·kg-1。六种彩叶植物矿质营养元素种类齐全,其中红花木的营养元素总量最高。六种彩叶植物均含有18种氨基酸成分,必需氨基酸和总氨基酸含量各不相同,但必需氨基酸占总氨基酸的比例却较为一致,且都高于40%。六种彩叶植物叶片内均含有一定量的铅、镉、砷、汞等有害元素,但有害元素尤其是砷和汞的含量均低于允许量。     

四个不同地理鲇群体肌肉营养组成的比较分析

为了丰富黄河、松花江和大洋河鲇的肌肉营养数据, 试验采用国家标准方法对4个自然河段鲇群体(黄河白银段BY、黄河郑州段ZZ、松花江哈尔滨段HEB和大洋河东港段DG)背部肌肉的营养组成进行测定。结果表明, BY群体粗蛋白显著(P0.05)高于ZZ和DG两群体, BY群体粗脂肪显著(P0.05)高于ZZ、DG和HEB三群体。四群体脂肪酸组成以不饱和脂肪酸为主, 单不饱和脂肪酸中油酸含量最高, BY群体油酸含量显著(P0.05)高于ZZ、DG和HEB三群体, BY群体EPA+DHA含量最低, 和ZZ、DG和HEB三群体有显著(P0.05)差异。四群体氨基酸组成均以谷氨酸含量最高。除缬氨酸和异亮氨酸外, 四群体其余氨基酸评分(AAS)均接近或大于1; 四群体各氨基酸的化学评分(CS)均大于0.5。缬氨酸为第一限制性氨基酸, 异亮氨酸为第二限制性氨基酸。必需氨基酸指数(EAAI), HEB群体最高, DG群体最低。该结果说明, BY群体与ZZ、DG和HEB三群体营养价值差别大, ZZ、DG和HEB三群体间营养价值接近。四群体的营养差异可能由不同地理环境导致。