糙皮侧耳Pleurotus ostreatus子实体凝集素基因polectin2及其启动子的克隆及功能鉴定

凝集素在食用菌的生长发育、抗病虫等逆境及抗肿瘤等方面发挥着重要的作用,但是关于糙皮侧耳凝集素基因的抗逆功能研究还比较薄弱。因此,本文利用PCR技术克隆得到糙皮侧耳子实体凝集素polectin2基因及其启动子序列,测序和生物信息学结果显示该基因不含内含子,完整开放阅读框为408bp,编码136个氨基酸,预测蛋白分子量15.3k Da;启动子polectinpro序列长度为927bp,生物数据库PLACE分析结果表明polectinpro含有低温、干旱胁迫诱导和病程相关等抗逆元件。然后,构建含有GFP融合蛋白的重组植物表达载体pCAMBIA1302-polectinpro-gfp,并进行烟草遗传转化,结果证明该启动子能够在低温诱导下驱动gfp基因表达。同时成功构建了含有polectin2的重组植物表达载体pCAMBIA1301-polectin2并进行了烟草遗传转化;对阳性转化植株T1代和野生型种子进行低温胁迫实验,结果表明转基因种子萌发率明显高于野生型。综上所述,推测polectin2基因具有耐低温功能,且其启动子具有低温诱导活性。上述结果为进一步利用子实体凝集素polectin2基因提高食用菌的耐低温等抗逆功能提供了理论依据和技术支持。     

Veratryl alcohol stimulates fruiting body formation in the oyster mushroom, Pleurotus ostreatus

The oyster mushroom, Pleurotus ostreatus, cultivated in solid state on sugarcane bagasse-wheat bran (5:1) medium in the presence of veratryl alcohol resulted in an increased production of the fruiting body at earlier times compared to when the fungus was grown in the absence of veratryl alcohol. The results indicate a new physiological role for veratryl alcohol in stimulating fruiting body formation. Veratryl alcohol also stimulated laccase production during the mycelial growth stage. Evidence is also presented that laccases were involved in the physiological development of the fruiting body.     

Purification, structure and immunobiological activity of a water-soluble polysaccharide from the fruiting body of Pleurotus ostreatus

The water-soluble polysaccharide (POP), with a molecular mass of 2.4x10(4)Da, was obtained from the fruiting body of Pleurotus ostreatus. Structure features of the purified polysaccharide were investigated by a combination of chemical and instrumental analysis, such as methylation analysis, Smith degradation, GC-MS, (13)C and (1)H NMR and FTIR. The results indicated that the backbone of POP was composed of (1-->6)-linked-alpha-D-galactopyranosyl and (1-->2,6)-linked-alpha-D-galactopyranosyl residues, which were terminated with a single terminal (1-->)-beta-D-glucose residue at the O-2 position of galactosyl along the main chain in the ratio of 1:1:1. Preliminary tests in vitro showed POP is capable of enhancing concanavalin A (ConA)- or lipopolysaccharide (LPS)-induced lymphocyte proliferation, which suggested that POP could be a potential immunostimulating agent for use in functional foods or medicine against both pathogens and cancer.     

Structure-activity relationships of the saponins dioscin and dioscinin

Hemolytic and antifungal activities of partial acid hydrolysates of dioscin and dioscinin were compared. In general, these activities were proportional to the number of sugar residues and those derivatives having branched sugar chains showed higher activities than those with straight chains. The 17-hydroxyl group of the dioscinin derivatives reduced both biological activities.     

Gastroprotective activities of a polysaccharide from the fruiting bodies of Pleurotus ostreatus in rats

In this study, a water-soluble polysaccharide (POPw) was successfully purified and identified from the fruiting bodies of Pleurotus ostreatus, with a molecular weight of 2.3×10(4)Da. POPw contained 97.1% total sugar, 0.3% uronic acid and 0.2% protein. Gas chromatography (GC) analysis suggested that POPw was composed of Glc (52.3%), Gal (25.8%), Man (10.0%), Rha (6.1%), and Ara (5.2%). Animal experiments showed that oral administration with POPw significantly inhibited acetic acid-induced gastric lesions in rats, accompanied with a significant increase in mucus synthesis and the prostaglandin production. In addition, POPw could significantly increase the level of glutathione (GSH) and the activity of superoxide dismutase (SOD), as well as decreasing the content of thiobarbituric acid-reactive substances (TBARS) in acetic acid-treated rats with gastric ulcer. These results suggested that the gastroprotective effects of POPw on mice ulcer models can be attributed to its ameliorating effect on oxidative damage and reinforcing effect of gastric mucosal resistance.     

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.     

Purification and characterization of a new ribonuclease from fruiting bodies of the oyster mushroom Pleurotus ostreatus.

A ribonuclease (RNase), possessing an N-terminal sequence disparate from those of ribonucleases from other mushrooms and previously isolated Pleuotus ostreatus RNases, was purified from the fruiting bodies of the edible mushroom Pleurotus ostreatus. The N-terminal sequence of Pleurotus ostreatus RNase did not manifest homology even to a previously reported RNase from the same mushroom. The ribonuclease was adsorbed on CM-Sepharose and Mono S. It exhibited a molecular mass of 12 kDa in both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and gel filtration on Superdex 75. The ribonuclease displayed an activity of 11490 U/mg on yeast tRNA. The highest ribonuclease activity was exhibited toward poly U, followed by poly A and poly C. No activity was shown toward poly G. The optimal pH for its activity was 7 and the optimal temperature was 55 degrees C. It inhibited cell-free translation in a rabbit reticulocyte lysate with an IC50 of 240 nM.     

Cultivation of Pleurotus ostreatus on weed plants

Oyster mushroom, Pleurotus ostreatus (Jacq.:Fr.) Kumm. ITCC 3308 (collected from Indian Type Culture Collection, IARI, New Delhi, India, 110012) was grown on dry weed plants, Leonotis sp, Sida acuta, Parthenium argentatum, Ageratum conyzoides, Cassia sophera, Tephrosia purpurea and Lantana camara. Leonotis sp. was the best substrate in fruit body production of P. ostreatus when it was mixed with rice straw (1:1, wet wt/wet wt) for mushroom cultivation. The fruiting time for P. ostreatus was also less on Leonotis sp. than on any other weed substrates tested in the present investigation. T. purpurea was the least suited weed for oyster mushroom cultivation. The main problem of oyster mushroom cultivation on weed substrates was found to be low yield in the second flush that could be overcome by blending weed plants with rice straw. The protein contents of the fruit bodies obtained from Cassia sophera, Parthenium argentatum and Leonotis sp. were not only better than rice straw but also from the rice straw supplemented weeds.     

Yellow Blotch of Pleurotus ostreatus

Yellow blotch disease of the oyster mushroom (Pleurotus ostreatus) was first observed in a commercial mushroom farm in California in 1983. The disease, caused by Pseudomonas agarici, is characterized by primordia, with yellow droplets on their surface, which become stunted, yellow to orange, and deformed as they mature.     

不同诱导培养基对糙皮侧耳液体发酵产漆酶活性的影响

利用1株糙皮侧耳Pleurotus ostreatus栽培菌株为材料,研究添加碱性木质素或者配合简单碳源或氮源后对其液体发酵产漆酶活性的影响。结果表明,不同诱导培养基对糙皮侧耳漆酶活性具有极显著的影响（P<0.001）,而且不同诱导培养基对糙皮侧耳菌丝生物量也产生了极显著的影响（P<0.001）。此外,只利用碱性木质素或者是再添加碳源葡萄糖均有利于糙皮侧耳产漆酶,既包括产漆酶酶活性的提升,同时也包括产漆酶时间的提前,但只利用碱性木质素诱导不利于菌丝生物量的积累;而富含简单碳/氮源的诱导培养基,无论是否含碱性木质素,均有利于菌丝生物量的积累,其中,富含简单碳/氮源的培养基中再添加碱性木质素后的菌丝生物量和漆酶活性均高于不添加碱性木质素时的菌丝生物量和漆酶活性。相比而言,含碱性木质素的培养基中测得的漆酶活性大部分时间下都要高于不含木质素的简单碳/氮源培养基,含碱性木质素的培养基对糙皮侧耳菌株产漆酶的诱导作用更强。     

Structural characterization and in vitro antitumor activity of a novel polysaccharide isolated from the fruiting bodies of Pleurotus ostreatus

A novel water-soluble polysaccharide (POPS-1) was obtained from the fruiting bodies of Pleurotus ostreatus by hot water extraction, ethanol precipitation, and fractionated by DEAE-cellulose ion exchange chromatography and sepharose CL-6B gel filtration chromatography using an ATKA explore 100 purifier. The structure characterization and antitumor activity of the POPS-1 were evaluated in this paper. According to GC analysis, HPGPC, FT-IR, partial acid hydrolysis, periodate oxidation and Smith degradation, methylation and GC-MS analysis, the results indicate POPS-1 (M(w)=31 kDa) was composed of Man; Gal; Glc with a molar ratio of 1:2.1:7.9, it had a backbone of beta-(1-->3)-linked glucose residues, which occasionally branches at O-6. The branches were composed of (1-->3)-linked Glc, (1-->4)-linked Gal, (1-->4)-linked Man, and terminated with Glc and Gal residues. Cytotoxicity assay showed POPS-1 presented significantly higher antitumor activity against Hela tumor cell in vitro, in a dose-dependent manner, and exhibited significantly lower cytotoxicity to human embryo kidney 293T cells than Hela tumor cells compared with 5-Fu. The results suggest POPS-1 may be considered as a potential candidate for developing a novel low toxicity antitumor agent.     

Growth characteristics of Pleurotus ostreatus in bioreactors

Pleurotus ostreatus was cultured in a bioreactor equipped with different impeller geometries under non-limiting nutrient conditions. With a Rushton turbine impeller the specific growth rate decreased 30% and pellet diameter was reduced 15% when the aeration rate was increased from 1 to 1.5 vvm. Agitation rate reduced the pellet diameter from 5.1 mm to 2.8 mm using 200 rpm and 400 rpm of agitation, respectively. Specific growth rates of 0.036, 0.020 and 0.041 h–1 were obtained with Roshton (disc turbine), Helical Ribbon and InterMIG impellers, respectively. Impeller geometry is important to control the pellet size and consequently growth rate of P. ostreatus.     

刺芹侧耳下脚料多糖的纯化与结构解析

采用超滤分离结合乙醇沉淀的方法,从刺芹侧耳下脚料水提物中纯化获得一多糖组分PEP30。苯酚硫酸法检测其糖含量约为94.2%,HPSEC-MALLS-RI系统分析其重均分子量(Mw)为3.74×106Da,多分散系数为1.03,为窄分布样品。通过红外光谱、单糖组成分析、甲基化GC-MS分析和核磁共振技术对多糖的结构特征进行了研究。结果表明,PEP30为一种β-D-葡聚糖,其主链以β-(1→3)-糖苷键连接,支链以β-(1→6)-糖苷键连接,支链与主链上糖残基的摩尔比为1:3。     

A novel peptide with ribonuclease and translation-inhibitory activities from fruiting bodies of the oyster mushroom Pleurotus ostreatus.

From the fresh fruiting bodies of the oyster mushroom a peptide with a molecular weight of 9 kDa and demonstrating a novel N-terminal sequence GPCYLVAFYESSGRR was isolated. The isolation procedure involved ion exchange chromatography on CM-Sepharose and Mono S. The peptide was adsorbed on both types of chromatographic media. The peptide demonstrated a ribonuclease activity of 650 U/mg toward yeast transfer RNA. It inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC50 of 15 nM.     

Examining genetic relationships of Chinese Pleurotus ostreatus cultivars by combined RAPD and SRAP markers

Combined randomly amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) were used to assess the genetic diversity of Pleurotus ostreatus strains cultivated in China. For the RAPD and SRAP analyses, 479 and 282 polymorphic bands were obtained from 20 P. ostreatus strains using 20 and 13 selected primers or primer pairs, respectively. A combined RAPD/SRAP dendrogram grouped the 20 strains into five clades with a coefficient of 0.690. The comparison of RAPD and SRAP was evaluated in the present study. The combined RAPD/SRAP markers provided reliable information regarding the relationships among the P. ostreatus strains.     

平菇多糖清除02-及对红细胞膜自由基氧化的影响

平菇经热水抽提,Sevag法去蛋白后,用乙醇沉淀,得平菇粗糖.上Sephadex G-100层析,可得精制多糖.在体外反应产生O2-,此体系中,加入不同浓度的平菇多糖.平菇多糖在较低浓度时(<200 mg/L)具有清除O2-作用,而在较高浓度(>200 mg/L)时,作用不明显.同时,用荧光法研究平菇多糖对小鼠红细胞脂质过氧化的影响,结果表明平菇多糖能够抑制小鼠红细胞的脂质过氧化.     

Pantoea beijingensis sp. nov., isolated from the fruiting body of Pleurotus eryngii

Four Gram-negative-staining, facultatively anaerobic bacterial isolates were obtained from a fruiting body of the edible mushroom Pleurotus eryngii showing symptoms of soft rot disease in Beijing, China. Partial 16S rRNA gene sequencing, together with partial rpoB sequencing, placed these isolates in the genus Pantoea. Multilocus sequence analysis based on the partial sequences of gyrB, rpoB, infB and atpD revealed Pantoea dispersa and Pantoea gaviniae as their closest phylogenetic relatives and indicated that these isolates constituted a possible novel species. DNA–DNA hybridization studies confirmed the classification of the new isolates as a novel species and phenotypic tests allowed for differentiation from the closest phylogenetic neighbours. The name Pantoea beijingensis sp. nov. [type strain LMG 27579^T = KCTC 32406^T = JZB2120001^T (deposited at Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences)] is proposed.     

平菇病毒dsRNA传播途径的探究

通过对平菇孢子dsRNA检测、单双杂交和单单杂交,从垂直传播和水平传播2个方面来研究平菇病毒dsRNA的传播途径,结果表明：病毒dsRNA通过孢子垂直传播的几率很小,通过细胞融合的水平传播几率相对比较高,在实际生产中平菇病毒dsRNA的主要传播途径还有待进一步的研究。