巴西蘑菇菌体深层发酵培养条件的优化及成分分析*

对巴西蘑菇深层发酵条件进行了详细的研究,确定了巴西蘑菇菌体发酵的最优方案是:7%葡萄糖,1.5%酵母膏,0.3%磷酸氢二钾,0.1%硫酸镁,VB1 10mg/100ml,自然pH,接种量为15%,装液量为100ml/250ml三角瓶,150r/min,25℃恒温培养9天,菌丝干重达到1.8g每100ml发酵液。通过对巴西蘑菇子实体与深层培养菌丝体中蛋白质营养成分及多糖含量进行了分析比较,发现它们均是极好的营养食品之源。     

基质碳氮比对广叶绣球菌生长发育的影响

培养基质碳氮比显著影响食用菌菌丝生长及子实体生长发育。广叶绣球菌是一种珍贵的食药用真菌,前期研究发现其生长发育过程中对碳源的需求较多。本研究采用松木屑、复合氮源作为栽培原料,研究基质碳氮比对绣球菌菌丝生长及子实体生长发育的影响。结果表明：6种不同碳氮比（C/N）的栽培基质对菌丝生长速度无显著影响,但是随着C/N比值的增加,子实体形成期逐渐延长,子实体形成率呈先增大后减小的趋势。当基质C/N在48.8-55.9之间时,采收率较高,在80-90d内,达到采收标准的菌袋比例分别为83.7%和81.4%。当基质C/N为43.4时,鲜菇产量最高,但外观品质受影响。综合考虑生产成本及效率,采用松木屑和复合氮源作为栽培原料、适宜的基质C/N为48.8时,绣球菌子实体形成期为64d,子实体形成率为95%且较为集中,鲜菇平均产量为231g/袋（76%集中于200-260g/袋）,整个生产周期为84d。本研究结果表明绣球菌具有“高碳低氮”的营养生理特性。     

Enzyme activities and substrate degradation during white rot fungi growth on sugar-cane straw in a solid state fermentation

Two strains of Pleurotus spp., grown in solid state fermentation on sugar-cane straw, degraded the dry matter by 50% after 60 days. The rate of substrate consumption and the dry weight of fruiting bodies decreased in consecutive flushings. Both strains vigorously attacked hemicellulose (80% of total degradation) and lignin (70%). Fruiting bodies were rich in protein and lipids, and had a low content of carbohydrates and ash.     

Production of γ-linolenic acid by Cunninghamella echinulata cultivated on glucose and orange peel

A newly isolated strain of Cunninghamella echinulata grown on glucose produced significant quantities of biomass and cellular lipids in media with high C/N ratio. The oil yield from glucose consumed increased after nitrogen exhaustion in the growth medium, but gamma-linolenic acid (GLA) content in cellular oil systematically decreased during the lipid accumulation process. When lipid accumulation was completed, GLA concentration in the cellular lipids progressively increased. The highest GLA production (720 mg/l) was achieved in medium with a C/N ratio equal to 163. C. echinulata was also able to grow on orange peel. The C/N ratio in the orange peel decreased from 50 to 26 during solid-state fermentation. Maximum oxygen uptake was observed during assimilation of reducing sugars, whereas a polygalacturonase activity was detected after reducing sugars had been exhausted. The maximum GLA production was 1.2-1.5 mg/g of fermented peel, calculated on a dry weight basis. After enrichment of the pulp with inorganic nitrogen and glucose, an increase in the production of oil and GLA was observed.     

Tea waste as a supplement for the cultivation of <Emphasis Type="Italic">Ganoderma</Emphasis> <Emphasis Type="Italic">lucidum</Emphasis>

Tea waste (TW) was investigated as a new supplement for substrate mixtures in Ganoderma lucidum cultivation. The effects of sawdust (S) based substrates supplemented with TW at the various levels (75S:25TW, 80S:20TW, 85S:15TW, and 90S:10TW) and Ganoderma lucidum strains on yield, biological efficiency (BE) and the chemical composition of fruiting bodies were determined in solid-state fermentation. Significant differences were found among substrates regarding yield and BE, while yield and BE of the strains were not different. The substrate formulations producing highest yield and BE were 80S:20TW (87.98 g/kg substrate and 34.90%) and 75S:25TW (82.30 g/kg substrate and 31%). Yield and BE of substrates containing TW were generally higher than that of the control (80sawdust:18wheat bran:1sucrose:1 CaCO₃). Nitrogen, potassium, iron, and manganese contents and C:N ratios of substrates were strongly correlated with yield. BE showed positive and significant correlations with potassium, iron and manganese. Moisture content, potassium, magnesium, calcium, iron, and zinc contents of the fruiting bodies were affected by both strain and substrate. It was concluded that TW can be used as a supplement for substrate preparation in G. lucidum cultivation.     

糖类和植物生长调节剂对冬虫夏草子实体人工培养的影响

为提高人工培养冬虫夏草子实体的产量,需要优化其培养参数。本实验测定培养基中糖类和植物生长调节剂对冬虫夏草子实体产量的影响。于大米小麦作为主要组分的培养基中接入冬虫夏草菌,在9~13℃下培养60 d,转入4℃培养。葡萄糖培养基中,冬虫夏草子实体干重是麦芽糖培养基中的7.6倍,出现菌丝和收获子实体的时间也比麦芽糖培养基中至少快2个月;蔗糖培养基中未获得子实体。不同种类和浓度植物生长调节剂对冬虫夏草子实体产量影响显著。于菌液中加入环磷腺苷、三十烷醇和玉米素的培养瓶均未发现菌丝生长。加入100μg/mL 6-苄氨基腺嘌呤的培养瓶可见菌丝生长,但未见原基分化。转入4℃下6个月后,与对照相比,加入100μg/mL吲哚乙酸、1μg/mL和100μg/mL吲哚丁酸、10μg/mL赤霉素、1μg/mL和10μg/mL乙烯利,以及1μg/mL 2,4-D的培养瓶中子实体干重均显著提高,其中加入1μg/mL吲哚丁酸和1μg/mL 2,4-D的培养瓶的子实体干重是对照的15倍。实验结果为优化冬虫夏草子实体人工培育提供了支撑。     

中国东北地区三种蜜环菌的生物学特性及奥氏蜜环菌人工培养

蜜环菌属Armillaria真菌具有较高的食药用价值。由于蜜环菌的生长发育过程较复杂,还未完全实现商业化栽培,野生资源的供应受到季节性和地域性的影响。本研究以采自东北地区蜜环菌属的3个菌株为研究对象,通过培养物的形态特征及分子标记确定菌株JG19016为奥氏蜜环菌A. ostoyae,菌株JG19017为高卢蜜环菌A. gallica,菌株JG19018为中国蜜环菌生物种C。奥氏蜜环菌JG19016最适生长温度为25 ℃,高卢蜜环菌JG19017的最适生长温度为22 ℃,中国蜜环菌生物种C JG19018则在22-25 ℃时菌丝生长速度最快;3个菌株最适pH为5-6。奥氏蜜环菌JG19016对葡萄糖和蔗糖利用率较好,高卢蜜环菌JG19017对葡萄糖利用率较好,中国蜜环菌生物种C JG19018对葡萄糖和淀粉利用率较好;蛋白胨对3个菌株促进作用最强,为最适氮源。培养基中加入VB₁,对3个菌株的菌丝生长均有明显的促进作用。奥氏蜜环菌JG19016菌丝生长的最优培养基配方为：葡萄糖20 g,蛋白胨3 g,磷酸二氢钾2 g,硫酸镁1.5 g,VB₁ 10 mg,琼脂20 g,水1 L。在木屑基质中培养,其配方的最优碳氮比为38:1,最佳木屑粗细比为3:1以上。出菇条件探索结果显示,菌丝及菌索长满菌袋(17 mm×33 mm×5 mm丝聚乙烯袋)需要50-60 d,之后在18 ℃、60%湿度和12 h散射光的环境中,10 d左右可观察到原基产生。增加菇房湿度到90%-95%,2-3 d可观察到1-3 cm的幼子实体,7 d左右菌柄和菌盖完全分化,10 d左右观察到菌盖展开。     

不同培养料和发酵次数栽培巴氏菇比较

分别以玉米秸秆和稻草为培养料栽培巴氏菇,对菌丝生长性状进行对比;同时,在我国传统发酵栽培方法的基础上,将三次发酵法与二次发酵法进行了对比。试验结果表明:利用玉米秸秆栽培的巴氏菇在发菌速度及子实体质量和产量上均优于稻草;培养料经过3次发酵更适宜巴氏菇生长。经过3次发酵的玉米秸秆培养料栽培的巴氏菇产量和生物学效率分别为7.7 kg/m2和27.5%,子实体粗蛋白含量44.89%,可溶性糖含量44.01%,18种氨基酸总量31.70%,明显高于2次发酵的玉米秸秆培养料及2次、3次发酵的稻草培养料。     

HPLC指纹图谱技术在灵芝组织分离试验中的应用

利用 HPLC 指纹图谱技术研究从同一灵芝子实体不同部位组织分离获得的菌株三萜化合物的差异。用常规组织分离方法获得不同菌株,在 A、B、C、D 四种培养基上进行出菇试验,运用HPLC指纹图谱技术获得各子实体三萜提取物 HPLC 指纹图谱,计算图谱间的相似度,分析三萜指纹的差异。结果表明,从子实体不同部位分离获得的四个菌株在同一培养基相同条件下培养获得的灵芝子实体的三萜指纹图谱相似度均大于0.99;同一部位分离获得的菌株在不同培养基相同培养条件下获得的子实体,其粗三萜 HPLC 图谱相似度均大于0.96;不     

HPLC指纹图谱技术在灵芝组织分离试验中的应用

利用HPLC指纹图谱技术研究从同一灵芝子实体不同部位组织分离获得的菌株三萜化合物的差异。用常规组织分离方法获得不同菌株,在A、B、C、D四种培养基上进行出菇试验,运用HPLC指纹图谱技术获得各子实体三萜提取物HPLC指纹图谱,计算图谱间的相似度,分析三萜指纹的差异。结果表明,从子实体不同部位分离获得的四个菌株在同一培养基相同条件下培养获得的灵芝子实体的三萜指纹图谱相似度均大于0.99;同一部位分离获得的菌株在不同培养基相同培养条件下获得的子实体,其粗三萜HPLC图谱相似度均大于0.96;不同的组织分离部位和不同的培养基对灵芝菌株三萜指纹图谱的影响均不显著。18号菌株在C培养基上培养获得的子实体三萜含量最高,上层菌肉为最优组织分离部位,C培养基为最优培养基。灵芝的三萜组成不受生长环境的影响,而生长环境会对其三萜化合物含量产生一定的影响。本文首次应用HPLC指纹图谱方法对灵芝组织分离获得的菌株的差异性进行了研究。     

利用原生质体紫外诱变技术选育耐高温香菇菌株

【目的】运用原生质体紫外诱变技术选育香菇耐高温新菌株。【方法】以香菇菌株18为出发菌株,紫外诱变处理其原生质体,通过47°C热激3 h后菌丝恢复生长的情况来筛选获得耐高温诱变株,测定18及其所有诱变株在木屑培养基中的恒温长速、高温长速以及恢复长速,并进行高温出菇试验。【结果】筛选得到57株耐高温诱变株,其中诱变株N6、N44和N24的综合性状较好。恒温长速、高温长速以及恢复长速与出菇性状具有相关性,恢复长速与出菇产量、单菇性状、耐高温能力呈正相关,可初步作为预测耐高温菌株综合性状的指标。【结论】利用原生质体紫外诱变技术,可初步选育出耐高温香菇新菌株。     

布莱克韦尔虫草的生物活性评价及其子实体的人工培育

虫草在世界范围内广泛分布,具有多种生物活性及较高的食药用价值。为进一步丰富虫草真菌资源,本研究对从广东韶关丹霞山采集到的一份野生虫草样品进行了菌株的分离鉴定、生物活性评价和子实体的诱导培养。通过形态学和系统发育分析,将菌株ZYJ0835鉴定为布莱克韦尔虫草Cordyceps blackwelliae。利用5种不同的培养基进行液体发酵培养,发现该菌株的发酵上清液和菌丝体均具有抗氧化和纤溶活性。此外,该菌株还具有一定的抑菌活性。利用大米、小麦、柞蚕蛹3种培养基质,均成功诱导出子实体,并且人工培养的子实体也具有抗氧化和纤溶活性。本研究首次在中国境内报道布莱克韦尔虫草的分布,为进一步开发利用该虫草资源提供了理论依据。     

Solid-state fermentation of carob pods for ethanol production

The production of ethanol from carob pods by Saccharomyces cerevisiae in solid-state fermentation was investigated. The maximal ethanol concentration (160±3 g/kg dry pods), ethanol productivity (6.7 ± 0.2 g/kg per hour), ethanol yield (40 ± 1.8%), biomass concentration (7.5 ± 0.4 x 10⁸ cells/g carob pulp) and fermentation efficiency (80 ± 2%) were obtained at an inoculum amount of 3%, a particle size of 0.5 mm, a moisture level of 70%, a pH of 4.5 and a temperature of 30°C. Under the same fermentation conditions both sterilized and non-sterilized carob pods pulp gave the same maximum ethanol concentration.     

Lignocellulolytic enzymes profile during growth and fruiting of Pleurotus ostreatus on wheat straw and tree leaves

Cultivation of two commercial Pleurotus ostreatus (oyster mushroom) strains was performed in plastic bags. Tree leaves appeared to be an excellent growth substrate for the conversion into fruiting bodies with biological efficiency of 108-118%. The level of enzyme activity was strongly regulated during the life cycle of mushrooms. However, despite the quantitative variations, each strain had a similar pattern of enzyme accumulation in fermentation of both substrates. Laccase and MnP activities were high during substrate colonization and declined rapidly during fruiting body development. On the contrary, in substrate colonization P. ostreatus expressed comparatively low activity of hydrolases. When primordia appeared, the activity of these enzymes sharply increased. Both cellulase and xylanase activity peaked at the mature fruiting body stage. When mushrooms shifted to the vegetative growth, the activity of ligninolytic enzymes again gradually increased, whereas the activity of hydrolases decreased.     

Xylanase production by Ganoderma lucidum on liquid and solid state fermentation

Ganoderma lucidum, a white rot fungus, was exploited for its potentials to produce xylanase employing shake and solid-state culture conditions. Different culture conditions such as pH, temperature, carbon and nitrogen requirements for its growth and production of xylanase were optimized. The culture media pH 6.0-7.0 and temperatures 30 degrees-35 degrees C significantly promoted the growth as well as xylanase secretion into the media. Xylan and peptone were found to be the suitable carbon and nitrogen sources. Among the different agrowastes used, wheat bran was found to be the best substrate for the test fungus for the production of xylanase than sugarcane bagasse and rice bran in solid-state fermentation.     

Some Factors Affecting Fermentation Capacity and Net Growth of Rumen Microorganisms

The fermentation rate of rumen microorganisms obtained from a sheep fed on hay was determined at different hours after feeding. The highest rate of fermentation was obtained immediately after feeding whereas the lowest rate was always before feeding. The effect of adding glucose, sucrose, and starch alone or in combination with urea, sodium citrate, or succinic acid on the maximal fermentation rate was studied. In no case was the maximal fermentation rate greater than that obtained in the presence of concentrate and straw. An investigation on the effect of C/N ratios, dilution, and substrate concentration on the net growth of rumen microorganisms was undertaken. The highest net growth value was obtained with a C/N ratio of 16.5 to 17.5, which is the ratio found for the ration fed to the animal. It was possible to increase net growth approximately threefold by using the proper dilution and fivefold by using the proper feed concentration.     

蛹虫草对锌的耐性与富集特征

在培养基内添加不同量的锌,研究其对蛹虫草子实体的形成、子实体和菌丝体生物量、子实体多糖含量和葡萄糖含量的影响,以及蛹虫草子实体和菌丝体对锌的富集能力。结果表明锌对上述各项都有影响。液体培养条件下,锌浓度在453906mg/L范围内可以促进菌丝体生长,锌浓度超过4077mg/L时,菌丝生长受到抑制。培养基锌的浓度在4077mg/L以下时,蛹虫草菌丝体锌的富集量随着液体培养基锌浓度的提高而提高。固体培养条件下,锌含量在226453mg/kg范围内可以促进蛹虫草子实体生长,并且在此含量范围内,蛹虫草子实体中葡萄糖含量较高。培养基锌含量在680906mg/kg时,子实体多糖含量较高。培养基锌含量在2038mg/kg以下时,蛹虫草子实体中锌的富集量随着培养基锌含量的提高而提高,在培养基锌含量为2038mg/kg时,子实体中锌的含量达到28570mg/kg(干重)。     

信息素信号通路基因在斑玉蕈生长发育过程中的差异表达

为了更清楚地了解斑玉蕈菌丝成熟、原基形成和子实体发育的过程,本研究对不同菌丝培养时期的栽培瓶进行出菇实验,并对其不同培养时期和生长发育关键时期的信息素通路基因进行差异表达分析,以期揭示信息素信号通路基因参与调节斑玉蕈菌丝的生长、子实体形成和发育的作用。研究结果表明：斑玉蕈菌丝培养40-80d过程中,子实体产量呈上升的趋势,说明菌丝的成熟程度对产量会产生重要影响。对斑玉蕈基因组中的信息素信号通路基因进行分析鉴定共获得了8个关键基因。信息素通路基因差异表达分析表明：在菌丝培养40-80d过程中,大部分信息素信号通路基因在第60天时表达量最高,其中ste20、cdc24和ste12上调了4-20倍,而在第80天出现下降。从菌丝恢复到扭结形成原基和子实体发育的过程中,大多数基因在原基时期表达量最高,其中ste20、cdc24、ste11和ste12表达量上调最为显著,在子实体成熟期这些基因表达量下降。因此,这说明在菌丝营养生长过程中,在第60天菌丝细胞增殖生长最为旺盛,而在第80天菌丝细胞基本停止生长,菌丝也逐渐达到成熟。同时,在菌丝生殖生长过程中,斑玉蕈持续地上调信息素通路基因表达使菌丝细胞不断地分裂增殖,从而使新生的菌丝扭结形成原基,其中ste3、ste20、cdc24、ste11和ste12基因可能对斑玉蕈菌丝细胞的分裂增殖和诱导子实体形成起到关键的作用。     

Use of various coffee industry residues for the cultivation of Pleurotus ostreatus in solid state fermentation

Studies were carried out to evaluate the feasibility of using coffee industry residues, viz. coffee husk, coffee leaves and spent coffee ground as substrates in solid state fermentation (SSF) to cultivate edible mushrooms Pleurotus. Eight strains of Pleurotus ostreatus and two strains of Pleurotus sajor‐caju were screened on a medium prepared from aqueous extract of coffee husk and agar. Based on best mycelial growth (9.68 mm/day) and biomass production (43.4 mg/plate in 9 days at 24°C), the strain P. ostreatus LPB 09 was selected for detailed studies. SSF was carried out using these substrates under different moisture conditions (45–75%) and spawn rates (2.5–25%). In general, although a 25% spawn rate appeared superior, the 10% spawn rate was recommended for all the three substrates in view of the process economics, as there was not any significant difference in the increase with 10 to 15%. The ideal moisture content for mycelial growth was 60–65% for coffee husk and spent coffee ground, and 60–70% for coffee leaves. The biological efficiency (BE), which is defined as the ratio of the weight of fresh fruiting bodies to the weight of dry substrate, multiplied by 100, and which indicates the fructification ability of the fungus for utilizing the substrate, was best with coffee husk. With coffee husk as the substrate, the first fructification occurred after 20 days of inoculation, and the biological efficiency reached about 97% after 60 days. When coffee leaves were used as the substrate, no fructification was observed even upon prolonged cultivation. With spent ground as the substrate, the first fructification occurred 23 days after inoculation and the biological efficiency reached about 90% in 50 days. There was a significant decrease in the caffeine and tannin contents (61 and 79%, respectively) of coffee husk after 60 days. It was remarkable to observe that caffeine was adsorbed onto the fruiting body (0.157%), indicating that it was not completely degraded by the fungal culture. However, no tannins were found in the fruiting body, indicating that the fungal strain was capable of degrading them. The results showed the feasibility of using coffee husk and spent coffee ground as substrates without any pre‐treatment for the cultivation of edible fungi in SSF, and provided one of the first steps towards an economical utilization of these otherwise unutilized or poorly utilized residues.     

壳聚糖和ε-聚赖氨酸处理对双孢蘑菇贮藏过程中品质的影响

以延长双孢蘑菇货架期为目标,探讨壳聚糖和ε-聚赖氨酸处理对采后双孢蘑菇在4 ℃贮藏过程中生理特性、营养品质和贮藏特性的影响。结果表明：与对照组双孢蘑菇相比,壳聚糖与ε-聚赖氨酸6:4复配溶液处理能够有效抑制双孢蘑菇表面微生物的生长和多酚氧化酶活性的增加,保持双孢蘑菇子实体较高的L*值和硬度,延缓双孢蘑菇的质量损失和细胞膜透性的升高,减少双孢蘑菇子实体的腐烂,保持较高商品率。在4 ℃条件下,壳聚糖与ε-聚赖氨酸6:4复配溶液对双孢蘑菇的保鲜性能最优,能够有效保持双孢蘑菇的商品品质和延长其贮藏时间。