检测大蒜病毒和产生脱毒蒜的方法

简述了目测汰毒法、指示植物法、酶联免疫吸附检测法 (ELISA)、逆转录聚合酶链式反应法 (RT PCR)、直接组织印迹免疫测定法 (DTBIA)等病毒检测技术 ,以及采用茎尖培养、花序轴离体培养、茎盘培养、茎盘圆顶培养等方法产生脱毒蒜的关键技术及其脱毒效果     

芳香族化合物对斑玉蕈菌丝生物量、漆酶活性及其转录水平的影响

芳香族化合物适当时间适当浓度添加到培养基中,可提高真菌漆酶活性,有助于增强其对木质纤维素的利用效率。为了增强斑玉蕈漆酶活性,本文研究了8种芳香族化合物对其酶活的影响及其与菌丝生物量的相关性。研究发现在无诱导物条件下,斑玉蕈漆酶活性和菌丝生物量相关系数r为0.9956,说明它们呈正相关,但是整个培养过程漆酶活性相对较低;供试的芳香族化合物对漆酶活性都有不同程度的诱导作用,其中添加0.1mmol/L的愈创木酚对斑玉蕈漆酶活性诱导作用最大,达到3倍以上,同时提高了斑玉蕈菌丝生长速度和菌丝生物量;而随着添加时间的延长,部分化合物对漆酶活性和菌丝生物量都产生不同程度的抑制作用,这可能因为化合物对菌丝毒性的延长导致菌丝生长变慢或死亡;进一步研究发现,斑玉蕈3个漆酶同工酶基因lcc2、lcc3和lcc4在诱导剂愈创木酚的影响下转录水平都不同程度地上调。研究结果表明诱导漆酶活性可以提高斑玉蕈菌丝生长速度和生物量,暗示可能通过提高漆酶活性的方法,提高斑玉蕈的培养基利用效率。     

食用菌病毒的研究进展

病毒是引起食用菌发生病害的重要病原之一,由于其具有潜隐性、不易辨认的特点,因而难以被人们所察觉,一旦发病就难以控制。在食用菌研究领域中,食用菌病毒使食用菌产量严重下降,引起的病害越来越受人们的重视,逐步成为该领域的研究热点。因此,本文主要针对食用菌病毒的结构与分类、危害、传播方式、检测方法以及食用菌病毒的脱毒技术等方面进行了综述,并对其存在的问题和前景进行了展望。这为食用菌病毒的检测、脱毒和无毒菌种的生产提供了理论依据,并为食用菌病毒的深入研究提供参考。     

氮源浓度对平菇菌丝生长和子实体形成的影响

本文报道了合成培养基中不同氮素水平对平菇菌丝生长和子实体原基产生的影响。平菇菌丝对基质中氮含量的要求较宽。在０．０２８－０．０９３％范围内生长都良好;但缺氮时严重抑制生长,菌丝也不发生扭结。实验结果还表明,硫胺素可能对氮素利用有促进作用,较高的起始氮浓度（０．０４７－０．０９３％）有利于子实体原基的形成。     

苹果脱毒苗与常规苗脂肪酸组成与抗氧化酶活性的比较研究

以甘肃陇东黄土高原3个主栽苹果品种嘎拉、秦冠、富士的12代继代培养脱毒苗与常规苗进行叶片类脂脂肪酸组成及抗氧化酶活性的比较研究。结果表明,苹果脱毒的膜脂过氧化程度降低,叶片类脂脂肪酸不饱和度提高。过氧化氢酶(CAT)和过氧化物酶(POD)活性升高,而超氧化物歧化酶(SOD)活性与常规苗无明显差异。     

食用菌对黄曲霉毒素B1的脱毒作用研究

目的 研究平菇、金针菇、黑木耳等七种食用菌对黄曲霉毒素B1（AFB1）的脱毒效果。方法 首先通过降解圈直径与菌落直径之比（Da/Dm）筛选出具有脱毒作用的株菌;然后通过AFB1残留量筛选出脱毒能力最强的菌株;最后将该菌株接种于含AFB1的玉米粉和大米中,考查其对粮食中AFB1的脱毒效果。结果 初筛发现平菇、黑木耳、金针菇对AFB1有较强脱毒作用,其Da/Dm分别为1.6±0.02、1.5±0.01、1.4±0.02;复筛发现黑木耳的脱毒能力最强,与AFB1共培养10 d后能清除88.16%的AFB1;进一步发现黑木耳对玉米粉和大米中AFB1有一定去除作用,清除率分别为62.4%和15.73%。结论 黑木耳对受AFB1污染的粮食作物有较好的脱毒作用,可用其控制食品与饲料中的AFB1。     

金针菇褐化病毒(FvBV)脱毒方法

【目的】评价5种不同脱毒方法对金针菇(Flammulina velutipes)菌株的脱毒效果,筛选出脱毒率高和脱毒后金针菇菌株菌丝生长速度、生物量、漆酶活力等性状改善明显的脱毒方法。【方法】以栽培金针菇菌株F-4889为研究材料,从菌丝体中提取大小约2.0 kb的病毒dsRNA,经RT-PCR鉴定该病毒为金针菇褐化病毒(FvBV)。采用菌丝尖端分离、原基组织分离、原生质体单核化、有性生殖和核迁移5种脱毒方法对金针菇菌株进行脱毒处理,利用dsRNA技术和RT-PCR检测脱毒效果。【结果】菌丝尖端分离脱毒后得到1株脱毒菌株;原基组织分离法未能脱毒;原生质体单核化脱毒法得到3株脱毒单核菌株和2株原单杂交脱毒菌株;有性生殖脱毒法获得脱毒孢子单核菌株23株和单孢杂交脱毒菌株8株;核迁移脱毒后得到5株核迁移脱毒菌株。脱毒率依次为25.0%、0、7.5%、57.5%和100%。脱毒菌株的菌丝生长速度、生物量、漆酶活力等均优于出发菌株、菌丝尖端和原基组织分离菌株。【结论】这5种方法中原生质体单核化、有性生殖和核迁移脱毒法脱毒效果较佳,均能有效脱除FvBV,脱毒率高,脱毒后菌株菌丝生长速度、生物量、漆酶活力等均明显提高。     

火菇属野生菌株发酵液对栽培菌株的化感效应

为探究火菇属Flammulina野生菌株发酵液对栽培菌株的影响,以火菇属野生柳生金针菇F. rossica F52、野生金针菇F. filiformis F39和栽培金针菇F. filiformis F47为供试菌株,采用固体平板培养和液体发酵的方法,对比了添加不同体积分数火菇属野生菌株发酵液对栽培菌株平板中菌丝长速、液体发酵中菌丝生物量、干质量及漆酶和淀粉酶活力的影响。结果表明：不同体积分数野生金针菇菌株F39发酵液对栽培菌株F47菌丝长速有抑制作用,对发酵液中菌丝生物量无显著影响,当添加发酵液体积分数为50%时可显著提高菌株F47发酵液中漆酶活力,当体积分数为40%时,可显著提高菌株F47发酵液中淀粉酶的活力（P<0.05）;添加不同体积分数的野生柳生金针菇菌株F52发酵液对菌株F47除淀粉酶外其他指标均影响显著,添加体积分数为30%的发酵液对栽培金针菇F47菌丝长速具有显著的促进作用,当添加发酵液体积分数为40%时,对菌株F47液体发酵中菌丝干质量和漆酶活力均有显著促进作用（P<0.05）。证明了化感效应不仅存在于属间,在同属下的种间也存在。     

布洛芬对脱毒甘薯叶节苗生根和生长的影响

将脱毒甘薯叶节苗基端于布洛芬液浸 1 0min然后扦插 ,能有效地促使薯苗生根 ,增加根数和根干重 ,提高根系活力 ;并能促使薯苗生长 ,茎叶干重和腋芽发育成的枝数增加 ,茎节繁殖系数提高。布洛芬的最适作用浓度为 1 0 0 0mg·L-1 。     

启动子替代构建糙皮侧耳漆酶高产菌株

POXC是糙皮侧耳合成最多的一种漆酶。应用启动子替代技术,用构巢曲霉的甘油醛-3-磷酸脱氢酶基因（gpd）启动子替代POXC基因的启动子,构建了超量表达POXC糙皮侧耳转化子。转化子中POXC基因表达量比出发菌株提高了0.72–3倍。在PDA平板培养、PD摇瓶培养和棉籽壳试管培养条件下,转化子漆酶活力显著提高,比出发菌株提高了1.5倍以上。用棉籽壳栽培,转化子菇产量比出发菌株提高了16.2%,培养料中木质素含量比出发菌株减少21%。结果表明,应用高效启动子替代能够显著提高糙皮侧耳漆酶基因的表达量、漆酶活力及其木质纤维素降解能力。     

Chemical Composition and Character Impact Odorants in Volatile Oils from Edible Mushrooms

The aim of this study was to investigate the chemical composition and the odor‐active components of volatile oils from three edible mushrooms, Pleurotus ostreatus, Pleurotus eryngii, and Pleurotus abalonus, which are well‐known edible mushrooms. The volatile components in these oils were extracted by hydrodistillation and identified by GC/MS, GC‐olfactometry (GC‐O), and aroma extract dilution analysis (AEDA). The oils contained 40, 20, and 53 components, representing 83.4, 86.0, and 90.8% of the total oils in P. ostreatus, P. eryngii, and P. abalonus, respectively. Odor evaluation of the volatile oils from the three edible mushrooms was also carried out using GC‐O, AEDA, and odor activity values, by which 13, eight, and ten aroma‐active components were identified in P. ostreatus, P. eryngii, and P. abalonus, respectively. The most aroma‐active compounds were C₈‐aliphatic compounds (oct‐1‐en‐3‐ol, octan‐3‐one, and octanal) and/or C₉‐aliphatic aldehydes (nonanal and (2E)‐non‐2‐enal).     

Saponin Stimulates Fruiting of the Edible Basidiomycete Pleurotus ostreatus

The addition of saponin from Quillaja to malt extract agar dramatically stimulated fruit body development of Pleurotus ostreatus. Higher concentration of Quillaja saponin added to the medium suppressed the growth of pilei of fruit bodies. The results indicate that the triterpenoid-saponin is a bioactive substance for fruiting of P. ostreatus.     

不同栽培基质对糙皮侧耳不同发酵方式下产漆酶活性的影响

为了比较糙皮侧耳栽培种在不同常规栽培基质上的漆酶活性,分析更适合糙皮侧耳生长的栽培基质,以1株糙皮侧耳Pleurotus ostreatus栽培菌株为研究材料,研究在固态和液态发酵条件下添加木屑、玉米芯和棉籽壳这3种栽培基质后,对其产漆酶活性的影响。结果表明,不同栽培基质对糙皮侧耳漆酶活性具有极显著的影响（P<0.001）;不同发酵方法对糙皮侧耳漆酶活性也具有极显著的影响（P<0.001）,仅第2天差异不显著。固体发酵与液体发酵条件下,糙皮侧耳在棉籽壳培养基上所检测到的漆酶活性均高于在木屑或者玉米芯培养基上,表明棉籽壳对提高糙皮侧耳漆酶活性的诱导能力更强。此外,糙皮侧耳在棉籽壳培养基上能够快速分泌漆酶,表明棉籽壳对缩短糙皮侧耳漆酶分泌时间的诱导能力更强。     

Study of laccase production by Pleurotus ostreatus in a 5 l bioreactor and application of the enzyme to determine the antioxidant concentration of human plasma

Aims: To achieve high laccase production from Pleurotus ostreatus in a bench top bioreactor and to utilize the enzyme for determination of the total antioxidant concentration (TAC) of human plasma. Methods and Results: Laccase production by P. ostreatus studied in a benchtop bioreactor was as high as, 874·0 U ml^?1 in presence of copper sulfate. The enzyme was used to replace metmyoglobin and hydrogen peroxide for the estimation of TAC in human plasma. The trolox equivalent antioxidant concentrations determined by the laccase‐based method and metmyoglobin method ranged from 1·63 ± 0·011 to 1·80 ± 0·006 mmol l^?1 and from 1·41 ± 0·004 to 1·51 ± 0·008 mmol l^?1 plasma, respectively. Conclusions: Pleurotus ostreatus produced high amount of extracellular laccase in a benchtop bioreactor. The enzyme can be used to assay TAC of blood plasma without the interference encountered with the hydrogen peroxide and metmyoglobin mediated assay method. Significance and Impact of the Study: Laccase production by P. ostreatus obtained in this study was the highest among all reported laccase producing white‐rot fungi. Moreover, an accurate laccase‐based assay method was developed for detection of TAC in human plasma.     

Water-in-oil microemulsions as the reaction medium for the solvent-sensitive yellow laccases

To determine the applicability of water-in-oil microemulsions for enzymatic conversions catalysed by yellow laccase from Pleurotus ostreatus (YLPO) D1 the following were studied: (i) the catalytic activity of the YLPO D1 in the oxidation of typical phenolic substrates: catechol (CAT), 2,6-dimethoxyphenol (DMOP) and 2,2^'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), at the appropriate pH optimum values in aqueous buffer solutions and in 62 mM bis-2-(ethylhexyl) sulfosuccinate sodium salt (AOT) in isooctane water-in-oil microemulsions; (ii) the effect of acetonitrile (ACN) on the kinetic parameters of DMOP oxidation catalysed by this laccase; (iii) the optimum conditions for the laccase catalytic activity in AOT in isooctane w/o microemulsions (w, laccase, AOT concentration); (iiv) the possibility of using the optimum water-in-oil microemulsions for the oxidation of aromatic alcohols (veratryl alcohol (VA) and benzyl alcohol (BA)) and the oxidative degradation of selected pollutants (3-chlorophenol, anthracene (ANT) and fluorene (FLU)).     

Water-in-oil microemulsions as the reaction medium for the solvent-sensitive yellow laccases

To determine the applicability of water-in-oil microemulsions for enzymatic conversions catalysed by yellow laccase from Pleurotus ostreatus (YLPO) D1 the following were studied: (i) the catalytic activity of the YLPO D1 in the oxidation of typical phenolic substrates: catechol (CAT), 2,6-dimethoxyphenol (DMOP) and 2,2^′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), at the appropriate pH optimum values in aqueous buffer solutions and in 62?mM bis-2-(ethylhexyl) sulfosuccinate sodium salt (AOT) in isooctane water-in-oil microemulsions; (ii) the effect of acetonitrile (ACN) on the kinetic parameters of DMOP oxidation catalysed by this laccase; (iii) the optimum conditions for the laccase catalytic activity in AOT in isooctane w/o microemulsions (w, laccase, AOT concentration); (iiv) the possibility of using the optimum water-in-oil microemulsions for the oxidation of aromatic alcohols (veratryl alcohol (VA) and benzyl alcohol (BA)) and the oxidative degradation of selected pollutants (3-chlorophenol, anthracene (ANT) and fluorene (FLU)).     

8个平菇栽培菌株的亲缘关系及培养特性研究

研究了8个平菇栽培菌株的亲缘关系及培养特性。结果表明:8个栽培菌株分别属于3个品种,根据子实体性状可划分为深色和浅色2个品系。其中菌株P8、P5和P1菌丝生长速度快,生长势强,产量高。     

Screening of Pleurotus ostreatus isolates for their ligninolytic properties during cultivation on natural substrates

Thirteen basidiospore-derived isolates of Pleurotus ostreatus f6 strain differing in the level of ligninolytic enzyme production and other characteristics (mycelium extension rate, colony morphology) from the parental strain were cultivated on natural substrates. Under these conditions ligninolytic enzyme activity, loss of organic mass, polycyclic aromatic hydrocarbons (PAHs) degradation and colonization of sterile and nonsterile soil were studied. The activity of ligninolytic enzymes was substantially higher in straw than in liquid culture, although the differences between the isolates were less pronounced on this substrate. Some of the isolates showed a very good ability to decompose the lignocellulosic substrate (straw) and a relatively high loss of organic mass was found after 50 days of cultivation in these strains. The original strain f6 and isolates B13 and B26 successfully degraded all seven tested PAH compounds present in experimental soil samples, but the higher or lower ligninolytic enzyme production of isolates tested had no substantial effect on the extent of the degradation. In our screening, six basidiospore-derivedisolates growing well in nonsterile soil were found, whichcould be suitable for the prospective biotechnological exploitation.     

Preparation and crossing of basidiospore-derived monokaryons –- a useful tool for obtaining laccase and other ligninolytic enzyme higher-producing dikaryotic strains of Pleurotus ostreatus

Laccase and other ligninolytic enzyme higher-producing dikaryons of Pleurotus ostreatus were obtained after crossing of compatible basidiospore-derived monokaryons selected from the parental basidiospore population on the basis of exceptionality in enzyme production, mycelium extension rate and/or colony morphology. As all detected changes in enzyme activity, mycelium extension rate, colony appearance and degradation of the polymeric dye Poly B411 were relatively stable after repeated testing, the dikaryotic isolates prepared in this way seem to be useful for the future biotechnological exploitation. No correlation between the colony appearance or the mating type and the enzyme activity or other characteristics tested has been found.