西藏产双孢菇(Agaricus bisporus)挥发性呈香成分初探

文章采用水蒸气蒸馏法提取来自西藏的3个双孢菇样品(Agaricus bisporus)的挥发油,以1个广州样品作为对照。通过加入薄荷醇作为参照标准物质的方法,应用气相色谱-质谱法结合NIST谱库检索研究了样品中的挥发性化学成分和相对含量,从4个双孢菇样品中分析鉴定了40个化合物。藏产编号为2796,709,694中的含量分别为43.10%,37.15%,31.34%,其中含量较高的挥发性成分依次为苯甲醇、糠醛、苯甲醛和柏木醇;广州样品0107号的含量为27.24%,其中除含量较高的苯甲醇、糠醛、糠醇外,尚有2-(1,1-二甲基乙基)-3-甲基-环氧乙烷,2-丙烯醇和马索亚内酯。研究结果表明菌种来源、产地气候、栽培方式会影响双孢菇的风味,为双孢菇的综合开发利用提供了理论依据。

Research of Holotrichia diomphalia Larvae Cultivation Technology with Agaricus bisporus Residues

[Objective] The aim was to explore key technology to cultivate Holotdchia diomphalia larvae with Agaricus bisporus residues and to provide, technologicar sup- ports for processing of Agaricus bisporus residues. [Method] In the research, fodder thickness, population density and residues under different treatments were set to re- search effects on Holotrichia diomphalia larvae. [Result] The optimal thickness of fodder was 25 cm and the optimal feeding density was 44-56 larvae per hectare. The dry residues were more suitable, compared with decomposed residues and corn bran powders, for cultivation of Holotrichia diomphalia larvae. [Conclusion] Cultivation of Holotrichia diomphalia larvae with Agaricus bisporus residues is a new method to make use of Agaricus bisporus residues and of significance for extension of agricul- tural circulation chain, increase of economic benefits and ecological benefits.

Whole-year Cultivation Techniques of Agaricus bisporus in Lianyungang District

Conventional Agaricus bisporus production is mainly concentrated in au- tumn and winter. Thus the Agaricus bisporus supply is not available throughout the year. Even worse, the price of Agaricus bisporus is cheap in boom season, reduc- ing the economic benefits; but the product is unavailable off season. The whole-year production of Agaricus bisporus will help to solve the problem and increase the eco- nomic benefits of farmers. In this paper, the whole-year cultivation techniques of Agaricus bisporus were summarized. There are a total of 3 seasons of Agaricus bisporus （spring, summer, autumn-winter） throughout the year. And thus the whole- year supply of Agaricus bisporus is achieved, the utilization of mushroom house is improved and the income of farmers is increased.

Standardized Cultivation Technology of Agaricus bisporus

Agaricus bisporus is a complete nutrition food, widely cultivated, and the demand on cultivation is general y lower, which advances Agaricus bisporus industry to certain extent. In order to improve yield of Agaricus bisporus, the research ruled the standardized cultivation technology and increased the yield by 20%, which can be an effective way for improving farmers’ incomes.

Molecular Cloning and Expression of Polyphenoloxidase Genes from the Mushroom, Agaricus bisporus

The polyphenoloxidase（PPO） is the key enzyme considered to be responsible for mushroom browning.By using homology cloning and rapid amplification of cDNA ends（RACE）,two new PPO genes and the corresponding cDNA were identified from the fruit bodies of Agaricus bisporus（AbPPO3 and AbPPO4,GenBank accession nos.GU936494 and GU936493,respectively）.The genomic DNA sequences of AbPPO3 and AbPPO4 are 2 080 and 2 189 bp in length,respectively,encoding putative polypeptides of approximately 66 and 68 kDa.The deduced amino acid sequences show characteristic features of two copper-binding domains conserved in the type III copper proteins including fungal polyphenol oxidases.Sequence comparisons indicate that AbPPO3 and AbPPO4 present 55.3% similarity to each other（48% identity）.We also obtained more than 1.5-kb long sequences upstream of the start codon of the AbPPO3 and AbPPO4 and recognized them as their respective putative promoters.Analyses of the two PPO promoter regions show that they contain abundant cis-acting elements which are probably responsible for anaerobic induction,light,wound,stress,and auxin response.Semi-quantitative RT-PCR results indicate that AbPPO3 and AbPPO4 were highly expressed in the mature fruit bodies and up-regulated after 2-d storage of mushroom.These results suggest that AbPPO3 and AbPPO4 may play roles in A.bisporus browning and pigmentation during development and postharvest storage and the elements in promoters may act as regulatory elements for the inducible expression of AbPPO3 and AbPPO4.The successful cloning and expression analysis of AbPPO3 and AbPPO4 warrant a further investigation on the structure and function of A.bisporus PPO which points to the possible targets for genetic manipulation.

Identification of commercial cultivars of Agaricus bisporus in China using genome-wide microsatellite markers

Agaricus bisporus is one of the most widely cultivated mushrooms in the world. Commercial cultivars are usually phenotypically alike and easy to be copied by isolating tissue cultures. This brings great challenges to distinguish different cultivars and to protect new varieties. Thus, techniques for the accurate identification of cultivars are essentially required. In this study, we accurately identified 11 commercial cultivars of A. bisporus released in China by using microsatellite(SSR, simple sequence repeat) markers. SSR markers were developed by mining the genome sequence. A total of 3 134 SSRs were identified, of which 1 490 SSRs were distributed in gene models, and 1 644 in the intergenic regions. A total of 17 polymorphic primer pairs were developed and SSR fingerprints were constructed for all the commercial cultivars. These SSR markers generated a total of 73 alleles, with an average of 4.29 per locus. Specifically, the primer combination of AB_SSR_2341 and AB_SSR_2590 could distinguish all the 11 commercial cultivars. The similarity coefficients of the 11 commercial cultivars were between 0.56 and 0.95 indicating that some of them were close related. Our results provide an efficient technique for the identification of A. bisporus cultivars in China, which can also facilitate the marker-assisted breeding in the future.

Screening on Chemicals for Preventing Mycogone peniciosa Magn in Agaricus bisporus

[ Objective ] The aim was to screen out chemicals with good control effect against Mycogone peniciosa Magn in Agaricus bisporus. [ Method ] M. peniciosa ofA. b/sporus was taken as the detection bacteria, and nine kinds of chemicals were conducted plate efficacy test in the laboratory. Mushroom house validation was carried out for the screened chemicals with strong inhibition to M. peniciosa. [ Result] Nine kinds of chemicals had obvious difference in control effects on M. peniciosa and effects on growth ofA. bisporus; 2 400 and 3 200 times dilution of 78% Boer MZ WP and 2 400 times dilution of 69% Fulishuang WP after casing material processing had good control effects on M. peniciosa and had no apparent adverse effects on mycelial growth and fruiting development of A. bisporus. [ Conclusion] The 2 400 and 3 200 times dilution of 78% Boer MZ WP and 2 400 times dilution of 69% Fulishuang WP are worth popularizing.

Morphological Features of Agaricus bisporus and Indoor Toxicity Test of Chemicals

Through separating and purifying diseased mushrooms infected by Mycogone perniciosa in Gaochun Area, Nanjing City, morphological features of the bacteria were identified, and toxicity of the bacterial strains was tested. The results showed that five bactericides in the experiment had obvious inhibitory effect on the growth of M. perniciosa, especially 50% Prochloraz manganese chloride complex WP 5 g/L had the best efficiency.

Effect of protease inhibitor from Agaricus bisporus on glucose uptake and oxidative stress in 3T3-L1 adipocytes

Objective:To explore the effect of the protease inhibitor from Agaricus bisporus(J.E.Lange)Imbach(AbPI)on glucose uptake and oxidative stress in 3 T3-L1 adipocytes.Methods:Adipocytes were differentiated and stained with OilRed-O staining to confirm adipogenesis.The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay,intracellular reactive oxygen species generation through flow cytometry,and morphologically through confocal microscopy using propidium iodide,4,6-diamino-2-phenylindol dihydrochloride,and 2’,7’-dichlorofluorescein diacetate dyes.The uptake of fluorescent glucose analog,2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy.Results:MTT assay showed that the cell survival rate was(28.00±3.00)%,(92.33±2.60)%,and(71.34±2.10)%in the presence of 2 mM H2O2,AbPI alone,and AbPI and H2O2 both,respectively,in comparison to the control.Oil-Red-O staining indicated that Ab PI enhanced adipogenesis.AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone,and showed insulinsensitizing effect in the presence of insulin,but failed to stimulate the uptake in the absence of insulin.Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon Ab PI treatment.Confocal microscopy showed that the damaged cell population rose to 3.50%,117.84%,and 261.50%in the presence of Ab PI alone,AbPI with H2O2,and H2O2 alone,respectively.Conclusions:The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.

Preliminary Study on Mixed Use of Agaricus bisporus Residue and Microbial Agent to Control Cucumber Root-knot Nematode

Single or mixed use ofAgaricus bisporus residue and microbial agent had better control effect on cucumber root-knot nematode disease in greenhouse. The results showed that the control effect of single use of A. b/sporus residue was 32.9% - 34.7%, and the yield increasing rate was 2.1% - 8.7% ; the control effect of mixed use ofA. b/sporus residue and microbial agent was 59. 1% - 63. 6%, and yield increasing rate was 2.6% - 3.5%. The experimental dose had no ad- verse effects on soil physical and chemical properties. The mixed use of A. b/sporus residue and microbial agent was one of the effective ways to control root-knot nematode disease.

Two Routes to Produce Chitosan from Agaricus bisporus

Two methods were used to produce chitosan by deacetylation of chitin which was extracted from Agaricus bisporus stipes.The first one gives chitosan 1 with low yield of 2.5%,degree of acetylation(DA)of 4%,molecular weight(MW)of 2.973×10^(5)(g/mol).The second route produces chitosan 2 with higher yield of 41%,degree of acetylation(DA)of 17.23%,molecular weight(MW)of 2.939×10^(5)(g/mol).Both chitosans were characterized by XRD,FTIR,^(1)H-NMR spectroscopy nuclear magnetic resonance of proton.The molecular weight(MW)was determined by size exclusion chromatography(SEC).Thermal analysis shows that both chitosans have moisture content lower than 10%.However chitosan 2 has the less ash%which is the quality grade for chitosan medical applications.Accordingly,fungal chitosan 2 could have potential medical and agricultural applications.

Bioinformatics analysis of metallothionein gene from <i>Agaricus bisporus, Ganoderma lucidum, Taiwanofungus camphorates</i>and <i>Paxillus involutus</i>

Metallothionein (MT) gene has an important role in the detoxification of toxic metals especially some heavy metals. Based on the published sequences of MT genes from Agaricus bisporus, Garnoderma lucidum, Taiwanofungus camphoratus and Paxillus involutus as a type of the edible or medical mushroom in NCBI database, their molecular structures, physiological functions and evolutionary relationship were analyzed using the bioinformatics methods. Results showed that AbMT, GlMT, TcMT and PiMT genes contained complete open reading frame (ORF) and their theoretical points of the last three encoding proteins were higher than 7.0. AbMT, GlMT, TcMT and PiMT were relatively rich in random coil and extended strand, but transmembrane helices and signal peptides were not found. 4 MTs were mainly localized in cell nucleus (over 60%) and their cellular functions might have some relation to central intermediary metabolism. Multiple sequence alignment indicated relatively high identity (more than 52%) and short genetic distances (lower than 0.900) among 4 MT nucleotide sequences. Abundant genetic diversity and strong codon bias were found based on the halotype diversity, average number of nucleotide differences, nucleotide diversity, effective number of codons, codon bias index and scaled chisquare. Simultaneously, we deduced that 4 MT genes during molecular evolution were under positive selection. The present study might provide basis for further investigation of MTs molecular mechanisms and genetic laws.

提高双孢蘑菇(Agaricus bisporus)生产效益的新技术

采用培养料一次性翻堆发酵，露地垄式菇床，播种后随即覆土，菇床加覆盖层栽培双孢蘑菇．它比同等营养水平的常规室外荫棚栽培产量增加２１７％，成本降低２７１％，利率达７４６％，是室外荫棚栽培的２８倍．

Accumulation Characteristics and Resistance to Heavy Metal Contamination of Agaricus bisporus Varieties in Shanxi Province,China

In order to evaluate the accumulation characteristics of heavy metals in the cultivars of Agaricus bisporus in Shanxi Province and to select the varieties with high resistances to heavy metal pollution, atomic absorption spectrophotometer and the atomic fluorescence spectrophotometer were used as the determination methods to compare the heavy metal contents and accumulation characteristics of 6 different varieties. The results showed that 3 heavy metal elements （lead, chromium and cadmium） were tested in the 6 varieties of A. bisporus , all of which were contaminated by heavy metals, but basically no beyond the national standards for food safety. In general, these varieties had less quality risk and could be eaten safely. The accumulation characteristics of heavy metals in these varieties were analyzed, and the varieties were screened according to the resistance to heavy metals pollution. The results showed that variety No. 1 and No. 6 were the varieties with strong comprehensive resistance to the 3 types of heavy metal pollution, and variety No. 3 was a species with weak comprehensive ability in resisting heavy metal pollution.

Study on the Accumulation of Heavy Metals in Varieties of Agaricus bisporus

The accumulation characteristics and resistance to heavy metal pollution of six varieties of Agar/cus b/spoms were studied. The results showed that arsenic, mercury and cadmium content could be detected in the six varieties, and all of the varieties were contaminated by heavy metals ,but they did not exceed the limits in the national standards for food safety. These varieties had a low quality risk and can be eaten safely. Varie-ties 1 and 6 were strongly resistant to the three kinds of heavy metals, while variety 4 was weakly resistant to heavy metal pollution.

Analysis of Bacterial Communities Present in Agaricus bisporus Phase Ⅱ Compost Using Amplified Ribosomal DNA Restriction Analysis(ARDRA)(Ⅱ)——Analysis of Bacterial Communities

Changes in the bacterial communities inhabiting Agaricus bisporus compost during Phase Ⅱ treatment were monitored using amplified rDNA restriction analysis(ARDRA).Sequence analysis data indicated that the bacterial communities associated with the compost samples were far richer in composition when determined by ARDRA compared with traditional methods based on bacterial isolation.Furthermore,the composition of the communities,especially in terms of the dominant bacteria during each of the four treatment stages(beginning of Phase Ⅱ,after pasteurization,5 days aeration,end of Phase Ⅱ,respectively),were very different.Restriction endonuclease digestion of mainly bacterial clones from four 16S rDNA libraries,C1,C3,C6 and C7,previously constructed on the basis of 16S rDNA fragments amplified from the four different stages of the Phase Ⅱ composting process,revealed the presence of unique microbial types.Thermophilic bacteria belonging to the Bacilli,and previously unreported strains of Trichococcus,Planococcus,Caryophanon,and subclass γ-Proteobacteria,were identified among the clones from the compost sampled during the aeration period(C6).Bacteria belonging to Thermus thermophilus and subclass α-Proteobacteria were detected in C1 and C7 composts sampled at the beginning and the end of Phase Ⅱ,respectively.Clones of some uncultured bacteria were also scored.

Design and experiment of a picking robot for Agaricus bisporus based on machine vision

Harvesting represents the crucial stage in the cultivation process of Agaricus bisporus mushrooms.An important way for the production process of Agaricus bisporus to reduce costs and increase income is to ensure timely harvest of Agaricus bisporus,reduce harvesting costs,and improve harvesting efficiency.There are many disadvantages in manual picking,such as high labor intensity,time-consuming work and high cost.In this study,a set of mushroom picking platform including climbing mechanism,picking robot,and control system was designed and developed.The picking robot consisted of a truss mechanism,an image acquisition device,a mushroom collection device,and a picking actuator.The profile picking actuator could realize the function of constant force clamping.An online size detection algorithm for Agaricus bisporus based on deep image processing was proposed.The algorithm included removal of abnormal noise points,background segmentation,coordinate conversion,and diameter detection.The precision picking system for Agaricus bisporus with coordinate compensation function controlled by Industrial Personal Computer was designed,and the visual control interface was developed based on Labview.Through the performance test,the reliability of machine vision recognition and the overall operating stability of the picking platform were verified.The test results showed that in the process of machine vision recognition,the recognition accuracy rate was higher than 92.50%,the missed detection rate was lower than 4.95%,the false detection rate was lower than 2.15%,and the diameter measurement error was less than 4.50%.The image processing algorithm had high recognition rate and small diameter measurement error,which could meet the requirements of picking operation.The picking platform’s picking success rate was higher than 95.45%,the picking damage rate was lower than 3.57%,and the picking output rate was higher than 87.09%.Compared with manual picking,the recognition accuracy rate of the picking platform was increased by 6.70%,the picking output rate was increased by 1.51%.The overall performance of the picking platform was stable and practical.

Computer Supported Sensory Profiling Analysis of Three Agaricus Cultivars

The sensory profile analysis is a commonly used method in the evaluation of directly consumed horticultural products （fruits, vegetables, grapes etc.）. The results of this type of analysis give an opportunity to distinguish the evaluated products from each other from a consumption point of view. In this paper, three Agaricus species （＂white button mushroom＂, ＂cream type＂ and ＂almond portobello＂）, were characterized by sensory profile analysis. The sensory attributes can have an influence on consumer product preference therefore it is essential to describe products, for example mushrooms. This is the first report which focus on describing the full sensory profile of these fresh mushrooms.

葛根提取物对双孢菇防褐变保鲜的影响

褐变是影响双孢菇保鲜过程中的重要反应,对双孢菇的品质变化起着至关重要的作用。为探究葛根提取物对双孢菇褐变及品质的影响,该实验采用葛根提取物通过浸泡、涂抹、与柠檬酸组合等方法处理双孢菇,测定双孢菇储存过程中失重率、总酚含量、丙二醛含量、相对电导率、多酚氧化酶(polyphenol oxidase,PPO)活性、过氧化物酶活性、质构参数和色差等指标,以表征处理效果。结果表明,采用葛根提取物涂抹能提高双孢菇的保鲜品质,柠檬酸对品质保持有不同程度的促进作用。在第10天,与对照组相比,葛根提取物和柠檬酸组合处理的双孢菇失重率降低41.26%,总酚消耗量降低16.90%,丙二醛含量上升幅度减少85.69%;同时有效抑制相对电导率增大和PPO活性,减缓质构参数的下降和褐变指数的上升,维持双孢菇表面颜色亮度。相关性分析表明,双孢菇保鲜过程中的质量损失与PPO酶促褐变和质构参数呈极显著相关(P<0.01),L*值的变化与褐变指数和PPO酶促褐变呈极显著相关(P<0.01)。该研究为探究葛根提取物对双孢菇保鲜的影响提供了参考,有助于开发更健康、可持续的双孢菇保鲜方法。

双孢蘑菇栽培基质二次发酵过程真菌群落结构变化分析

为明确双孢蘑菇栽培基质二次发酵过程中微生物动态和多样性,采用高通量测序技术,测定了双孢蘑菇栽培基质的二次发酵过程中真菌群落结构变化,并探究群落组成与环境因子的关系。结果表明:双孢蘑菇栽培基质二次发酵过程4个发酵阶段真菌分类单元(OTU)总数999个,包含10个门、26个纲、56个目、126个科、240个属和403个种,其中嗜热链球菌属(12.72%~42.63%)、unclassified_f__Sordariaceae(0.12%~31.63%)、念球菌属(1.30%~17.32%)、曲霉属(2.57%~8.98%)、青霉属(2.88%~9.76%)和丝壳属(0.89%~4.18%)等属真菌占主导地位。发酵过程中,嗜热链球菌属、念球菌属、曲霉属、青霉属等属丰度在发酵的早、中期呈上升趋势,后期下降;嗜热链球菌属和青霉属真菌相对丰度于发酵中期达到高峰。氨气排放与堆体温度为发酵终点关键指标,冗余分析真菌群落与氨气排放和堆体温度的相关性表明,嗜热链球菌属、青霉属和念球菌属丰度与发酵温度和氨气排放呈正相关,其丰度越高,二次发酵周期短且发酵质量优。