射频辅助提取香菇柄多糖

探讨射频技术在香菇柄多糖提取中的应用,优化了射频辅助提取香菇柄多糖的工艺,并与传统水提法、超声波辅助提取法和微波辅助提取法进行了比较。采用Draper-Lin中心组合设计和全因素中心组合设计对影响射频提取的处理时间、料液比、作用温度、预浸泡时间进行了筛选和优化。优化结果为:射频处理5.2min,作用温度(82.6±3)℃,料液比1:40(5g:200mL),预浸泡时间20min。在此条件下,香菇柄多糖提取率达到43.1mg/g。与传统水提法、超声波辅助提取法和微波辅助提取法相比,得率分别提高了94.1%,40.8%和5.38%;与传统水提法相比,多糖纯度有所提高(42.8%→46.7%),提取时间也大大缩短(6h→5.2min)。     

氮离子束注入诱变筛选万隆霉素高产菌株的研究

为获得万隆霉素高产菌株,用氮离子束注入万隆霉素产生菌进行诱变处理,并用链霉素抗性筛选法和抑菌圈筛选法进行突变株的筛选。结果表明:通过能量为20keV、剂量为90×2.6×1013N+/cm2的氮离子束注入处理万隆霉素产生菌的分生孢子,筛选出万隆霉素高产菌株WN939,其摇瓶发酵产素水平较出发菌株提高了82.10%。经5代传代培养,其产素水平稳定。     

胡枝子栽培香菇试验

利用豆科多年生落叶灌木胡枝子的木屑做培养基主料栽培香菇,所产香菇味美可口,营养丰富,符合卫生标准。一亩水土流失山地栽植胡枝子800株,第二年便可产干枝条500公斤。用以栽培香菇,可产鲜菇约500公斤,获纯收入约650元。这不仅大大提高群众种植胡枝子的积极性,促进治理水土流失,而且也为水土流失地区找到了一条脱贫致富的新门路。     

胶质芽胞杆菌及其突变株生理特性的初步研究

本文对经过筛选培育出的分解硅酸盐能力较强的胶质芽胞杆菌突变株 UV0 9K及抗高温、耐酸碱菌株 p Ht10 K的生理特性进行了研究。其中 ,最佳氮源为麦麸汁 ,与出发菌株相比 ,分解硅酸盐能力分别提高 164%、12 .7%。最佳碳源为葡萄糖 ,与出发菌株相比 ,产酸能力分别提高2 60 %、155%。金属离子、培养温度、p H、气 /液比等也对突变株的生长有一定影响。其结果可作为选定工业化条件的参考。     

香菇多糖对鲤鱼（cyprinus carpio）离体培养免疫细胞的活性调节作用*

采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖（APS）和香菇多糖（LNT）后,分别采用 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响;实时定量PCR法测定头肾巨噬细胞细胞因子IL-1β诱导表达的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧爆发活性;其低浓度时对细胞的氮呼吸爆发活性无显著影响,随着作用浓度的增加表现为高浓度抑制作用;香菇多糖可以显著促进鲤鱼外周血白细胞的增殖,且能显著增强IL-1β的体外诱导表达。结果表明香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是一种有潜力的鱼用免疫增强剂。     

固氮粪产碱菌（Alcaligenes faecalis）胞外多糖突变株的筛选及特性

应用Ｔｎ５转座子对粪产碱菌野生型菌株Ａ１５０１进行诱变处理，用荧光增白剂及ＴＴＣ为标记筛选，分别获得胞外多糖缺陷型和丰富型突变株。激光共聚焦观察证实，ＥＰＳ突变株确实具有与野生型不同的表面特性，贴根试验表明，突变株贴根菌数均少于野生型，胞外多糖生成过多或过少均影响菌体与根表的有效结合。     

常压室温等离子诱变选育高产橙、黄色素红曲菌研究

为了提高红曲菌(Monascus sp.)产橙、黄色素的性能,采用新型的常压室温等离子体(ARTP)诱变技术处理红曲菌WM951孢子悬液。结果表明,最佳照射时间为90 s,此时红曲菌突变效果最好,其孢子致死率为78%,形态突变率为14.2%。试验最终从15株突变菌株筛选得到1株高产橙、黄色素的突变菌株WM951M1,对该菌株的发酵产物进行色价检测分析,发现该突变菌株产橙、黄色素能力比出发菌株分别提高136%、43%,30℃培养10d后其固态发酵红曲米中橙、黄色素色价最大值分别为3 620、3300 U·g-1。出发菌和突变菌所产色素的全波长扫面图谱显示,突变菌所产色素最大吸收峰出现在465nm处,并在410 nm处也有吸收峰,说明该菌主要合成橙、黄色素。在传代培养的过程中,该突变菌株表现出良好的遗传稳定性及形态稳定性,不仅可应用于生产橙、黄色素,也可用于生产色调偏橙型红曲米,还能丰富红曲菌菌株库资源。     

产黄纤维单胞菌CR-14菌株诱变选育及发酵条件优化

为提高产黄纤维单胞菌CR-14纤维素酶活力,对菌株进行亚硝酸、紫外线及复合诱变处理,筛选产纤维素酶活较高的突变株,并对其发酵条件进行优化。结果表明,经亚硝酸和紫外线复合诱变得到一株产纤维素酶活较高的菌株Y-UA-18,其酶活力为10.57 U,为原菌株产酶活力的1.67倍。通过正交试验得到菌株Y-UA-18产酶最佳培养基为:秸秆粉1.0%、复合氮源0.6%、KH2PO40.1%、Mg SO40.1%、Na Cl 0.08%;最佳培养条件为:初始p H值7.0、培养温度30℃、发酵时间3 d。通气量对菌株Y-UA-18产酶影响不明显,但在厌氧条件下发酵液酶活显著降低,0.1%TW-80对菌株Y-UA-18的产酶没有影响。     

离子注入对苹果酒酵母菌的影响

以野生型苹果酒酵母菌Y0 2 为出发菌株 ,利用 8× 1 0 15ion cm2 注入剂量对出发菌株Y0 2 进行诱变处理 ,得到 1株形态特征、部分生化特征、菌体蛋白质等发生明显变异的突变株IONⅡ 1 1dry ,该突变株的产酒率比出发菌株提高 2 2 4% ,表明离子注入诱变育种是行之有效的     

链霉菌Snea253菌株的变异分析及对根结线虫活性影响

为明确委内瑞拉链霉菌(Streptomyces venezuelae)自发突变对南方根结线虫活性的影响。本研究利用继代培养法选育Snea253突变株,对形态分化及相关基因进行分析,并测其代谢产物的生物活性。获得6株菌落形态差异大的自发突变菌株,其中不产色素的3株突变菌株Snea253-G、Snea253-B和Snea253-F杀线虫活性分别为61.0%、45.5%和35.0%;产色素的3株突变株Snea253-A、Snea253-H和Snea253-J杀线虫活性分别为48.7%、37.2%和37.9%。ssgA、bldC、bldD、whiB、whiE和whiI基因克隆测序结果显示各突变株基因序列没有差异,可能此6个基因没有直接参与控制Snea253菌株的自发突变。链霉菌自发突变不但使其形态发生变异,而且也影响了其代谢产物的生物活性,研究为从形态上筛选优良菌株提供了相关理论依据。     

Biodegradation of oak (Quercus alba) wood during growth of the shiitake mushroom (Lentinula edodes): a molecular approach

The chemical transformations that occur during growth of the shiitake mushroom (Lentinula edodes) on oak (Quercus alba) were investigated to improve mushroom cultivation and utilization of the spent substrate. Oak logs were decayed by L. edodes over 8 years, during which time they were sampled at six intervals (30, 40, 66, 76, 77, and 101 months). Fresh and decayed oak samples were analyzed using solid-state (13)C NMR and pyrolysis-gas chromatography-mass spectrometry as well as off-line thermochemolysis with tetramethylammonium hydroxide. Degraded oak exhibited lower carbon contents and increased oxygen content compared to the control. Solid-state (13)C NMR analysis revealed that polysaccharides were the major component of both fresh and decayed oak but that L. edodes mediated the preferential loss of cellulose and xylans as compared to lignin, which remained in an altered form. Several trends point toward the degradation of lignin, including a decrease in the proportion of syringyl units as compared to guaiacyl units and a reduction in side-chain length. An increase in guaiacyl and syringyl acid-to-aldehyde ratios occurred with growth, which suggested that the fungus had caused oxidation of Calpha-Cbeta bonds. The overall effect of L. edodes on oak is similar to that of many white-rot fungi, which simultaneously degrade all cell wall components.     

果蔬微孔膜气调包装模型与试验验证

建立微孔膜气调包装的理论模型是保证气调包装质量、进行微孔膜气调包装系统设计的关键。基于孔径与分子平均自由程的关系,应用Fick扩散定理,结合果蔬呼吸速率模型,建立微孔膜果蔬气调包装内外气体交换的数学模型。以香菇为对象,利用密闭系统法确定其呼吸速率;采用硬质罐加PE膜封合包装香菇并在25℃、50%RH下储藏50 h以上,测定不同初始气体组分、微孔孔数与孔径条件下包装内气体浓度变化,对香菇微孔膜气调包装模型进行试验验证。结果表明,模型预测值与试验数据吻合度较高,所建模型能较准确地预测香菇微孔膜气调包装内外气体交换过程。     

Basic composition and amino acid contents of mushrooms cultivated in Finland

The basic composition (moisture, total carbohydrates, dietary fiber, crude fat, ash, nitrogen, and protein) and amino acid contents were determined in the cultivated mushrooms Agaricus bisporus/white, Agaricus bisporus/brown, Lentinula edodes, and Pleurotus ostreatus. In addition, nitrogen-to-protein conversion factors were calculated for each species by dividing the sums of amino acid residues with total (Kjeldahl) nitrogen contents. The dry matter contents of mushrooms varied from 7.7% to 8.4%. The dry matter of mushrooms contained large amounts of carbohydrates, from 4.5 (A. bisporus/white) to 5.8 g/100 g fresh weight (L. edodes). L. edodes proved to be an especially good source of dietary fiber (3.3 g/100 g fresh weight); the other mushrooms contained 1.5-2.4 g/100 g fresh weight. Crude fat, ash, and protein (based on amino acid analysis) contents of the mushrooms varied 0.31-0.35, 0.49-0.78, and 1.8-2.09 g/100 g fresh weight, respectively. Mushrooms proved to be good sources of almost all essential amino acids when compared with common vegetables. The mean nitrogen-to-protein conversion factor analyzed in the present study was 4.7 +/- 0.21. When using this factor, a very good estimation of protein contents could be obtained for the main species of mushrooms cultivated in Finland.     

食用菌栽培相关木霉种的鉴定

从木霉污染的食用菌菌筒和子实体中分离纯化了49株木霉菌株。采用形态学方法以及ITS/5.8S测序分析,对这些木霉菌进行了分类鉴定。结果表明,中国福建、浙江等省食用菌栽培相关木霉种以哈茨木霉Trichoderma harzianum Rifai和长枝木霉T. longibrachiatum Rifai为主,少量为深绿木霉 T. atroviride Karsten和棘孢木霉T. asperellum Samuels;木霉污染菌的种类与采集地点、食用菌的种类有一定相关,如在浙江庆元香菇Lentinus edodes (Berk.)sing菌筒中分离的木霉主要是哈茨木霉,在广州刺芹侧耳（杏鲍菇）Pleurotus enyngii菌筒中分离的木霉主要是棘孢木霉,而从福建浦城食用菌污染袋中分离的木霉主要是深绿木霉。两种分类方法对木霉鉴定结果基本一致,因此形态学分类方法结合分子生物学方法使食用菌栽培相关木霉种的分类鉴定更准确和可靠。     

Effects of management on the yield and high-molecular-weight polysaccharide content of shiitake (Lentinula edodes) mushrooms

Shiitake (Lentinula edodes (Berk.) Pegler) mushroom production in the United States has increased greatly over the last twenty years. Additional expansion of the shiitake mushroom market should be possible if the product can be marketed as a functional food, i.e., a food that has health-promoting effects beyond its nutritional value. High-molecular-weight polysaccharides (HMWP) including lentinan in shiitake may promote human health. This study was conducted to determine if management protocols influence the HMWP of shiitake mushrooms. Results indicate that measuring the total carbohydrate content of water-extractable, ethanol-insoluble polysaccharides was a simple way to estimate HMWP. Results also indicate that log-grown shiitake contained more HMWP than did substrate-grown shiitake. Among log-grown shiitake, both mushroom strain and tree species influenced HMWP content. The results suggest that there is considerable variation among shiitake mushrooms in HMWP content and that production protocols influenced the HMWP content of mushrooms.     

固氮斯氏假单胞菌铵载体amtB基因的功能和结构分析

固氮斯氏假单胞菌(Pseudomonas stutzeri)A1501是分离自水稻根际的联合固氮细菌,属变形细菌γ亚族。全基因组序列分析表明该菌含有2个紧密连锁的的铵载体蛋白编码基因amtB1和amtB2,位于同一个操纵子中glnK基因下游。铵载体蛋白AmtB1和AmtB2分别包含12和11个跨膜结构域。为研究amtB的功能,构建了amtB极性双突变株(PKOB1)。amtB极性双突变导致菌株生长缓慢;在铵浓度分别为3.3和10mmol/L条件下,其固氮酶活仍保持在0.01mmol/L的最佳固氮条件的87%和74%,而野生型已经基本尚失固氮能力。以硝酸盐为唯一氮源时,突变株细胞培养液的铵浓度可达到3.4mmol/L,而野生型并未检测到铵,表明AmtB可能参与了P.stutzeriA1501的铵离子转运,并与固氮酶活和氮代谢相关。     

香菇分段变温红外喷动床干燥工艺参数优化

为获得节能低耗和品质较优的干香菇,利用红外喷动床干燥设备,对香菇分段变温干燥工艺进行试验研究。在单因素试验基础上运用Box-Behnken Design优化试验,研究前期风温、转换点含水率和后期风温对单位能耗、粗多糖含量、亮度L*值和收缩率的影响,通过加权综合评分法推导多项式回归模型,优化红外喷动床干燥工艺参数。经响应面优化的干燥参数为：前期风温56.00 ℃、转换点含水率53.00%、后期风温72.00 ℃,该工艺下单位能耗为143.52 kJ/g、粗多糖含量9.98 mg/g、亮度L*值68.11、收缩率83.15%,综合评分值为35.37,与预测值拟合度高达99.27%,表明应用红外喷动床干燥取得的香菇制品能满足当前香菇干燥的发展趋势及需求,为香菇干制品的综合应用及生产加工提供参考。     

香菇表达序列标签(EST)的SSR分析及EST-SSR引物的筛选

从真菌基因组计划网站(FGP)和NCBI网站数据库下载了总长度为8.1Mb的11150条香菇的EST(包括香菇的10条cDNA)序列，通过SSRhunter 1.3软件结合手工查找，从中发现2.83％即316条EST含有一共469个SSR.，平均每17.3kb出现一个EST-SSR。在所有EST-SSR中，三碱基和六碱基SSR出现最多，分别占EST-SSR总数的38.00％和20.00%，出现较多的基元为(A)n、(T)n、(GA)n、(AG)n、(TGA)n、(GAT)n和(TCTTT)n，占所有EST-SSR的35.39％。利用Oligo 6.0软件设计了51对EST-SSR引物，并选用其它物种的24对引物，对香菇菌株进行了PCR扩增。实验结果表明，设计的51对引物有39对具有明显的扩增产物，选用的其它物种的24对引物均没有清晰可见的特异性扩增产物。部分有扩增产物的EST-SSR引物具有多态性。