Two strains were selected as test materials from 28 strains of Stropharia rugosoannulata at home and abroad using SRAP molecular marker technology and high temperature progressive cycle domestication. Protoplast regeneration and fusion technology were applied for preliminary selection of thermotolerant strains, and 2 excellent strains, S20 and R10, were screened out. Thermotolerant mycelium and high yield feature S20 and high proportion of good quality mushrooms and thermotolerant fruiting body feature R10. Monospore hybridization was conducted between S20 and R10. After several subcultures and cultivation experiments under high temperature conditions, J8L4 (Shannongqiugai No. 3) was eventually selected as the best strain with thermotolerance and high yield. The strain could well grow under the compost material temperature of 27-34°C and air temperature of 40-47°C for 7 days consecutively without contamination and fruits in 40 days. The production reached 14.56kg/m 2, with biological conversion rate of 91.03% and good quality mushroom ratio of 66.21%. The results indicate that the multi-level domestication and screening using comprehensive multiple breeding techniques make the selected strain property and genetic stability better.
Insertion site analysis is very important in functional genomics research of Flammulina filiformis. The analysis methods commonly used are reverse PCR, thermal asymmetric interlaced PCR, Tail-PCR, chromosome walking, etc., and these methods are hard to operate, time-consuming, poorly specific and inefficient. In recent years, the next generation sequencing has been applied in identification insertion site that transformants were sequenced one by one, but it has a large workload and a high cost. In this study, a matrix design was used to mix the DNA of multiple transformants to form a sample pool. After resequencing, the insertion sites were analyzed. The sequencing data of M sample pools can analyze the insertion sites of M× (M+1)/2 transformants. The matrix design was used to detect 21 transformants, and 21 insertion sites were identified. This method indicates that matrix design is practicable for insertion site identification and suitable for large sample analysis, such as mutant library.
Cytochrome c peroxidase (CcP) is one of the major antioxidant enzymes involving in hydrogen peroxide degradation and plays important roles in oxidative response of fungi. A cytochrome c peroxidase encoding gene (ffccp) was obtained, based on the genome data of Flammulina filiformis. The ffccp gene sequence was 1 913bp in length, containing a complete open reading frame (1 098bp) and encoding 365 amino acids. The bio-informatic predict results showed that FfCcP likely located in “membrane bound mitochondria” but contained no transmembrane helices, signal peptides, or disulfide bridges. The sequences alignment results showed that FfCcP contained both the heme distal and proximal residues and peroxidase domain. The phylogenetic analysis showed that the FfCcP sequence was highly similar to Pleurotus ostreatus and other mushroom CcPs (identity>70%) and clustered into CcP clade. These results demonstrated that the protein encoded by ffccp was cytochrome c peroxidase. The RT-qPCR results suggested that ffccp gene expression was up-regulated by both oxidative stress and injury stress, but the regulatory mechanisms might be different. The gene expression of ffccp was also detected during fruiting body development, and the results showed that ffccp was highly expressed in the stipe of elongation stage. Further study also demonstrated that the ffccp gene expression pattern was strongly correlated with the stipe elongation rates. It seems that the high-level expression of ffccp may be beneficial for stipe elongation in F. filiformis.
As an inevitable natural factor, magnetic field has an important influence on the growth and development of organisms. However, there are few reports about the effect of magnetic field on large fungus growth. The growth and gene expression of Flammulina filiformis mycelium were detected by rubidium magnet treatment. The results showed that magnetic field of rubidium magnet inhibited the growth of Flammulina filiformis mycelium, and the inhibition was strong when the magnetic induction intensity was over 0.003T. A gene encoding the magnetic receptor named as ff-magr and an iron transporter ff-fief of Flammulina filiformis were obtained by homology matching. Quantitative PCR results showed that the expression of ff-magr was significantly down-regulated at both N and S poles, and the magnetic fields at both N and S poles had no significant effect on the expression of ff-fief, indicating that the magnetic receptor protein was involved in the response process of the inhibition of magnetic field, and the magnetic receptor not merely regulated the growth and development of mycelia by regulating iron content. The molecular mechanism of magnetic field influencing the growth and development of fungi needs further study.
Twenty-eight strains of Flammulina filiformis from China and abroad were used for genome resequencing. In total, 1 241 583 SNP and 623 670 InDel sites were detected by mapping with reference genome. The screened 1 474 high-quality SNP sites with PIC (polymorphism information content) ranged from 0.101 to 0.966 were applicated for further analysis on genetic distance, UPGMA phylogenetic trees, population structure, and PCA between strains. The result showed that genetic distance between strains were 0.057-0.631. UPGMA tree showed the cultivation strains was a subclade within a large mixed clade of wild and cultivation strains, and seasonal cultivation and industry cultivation strains can be distinguished, consisting with the history of breeding cultivation of F. filiformis. Five subpopulations were detected by STRUCTRUE software. The PCA result were consistent with the analysis based on other mathematical methods.
Using the parental monokaryons APP7 and M2S16 of the Auricularia cornea hybrid APM2-16 as research materials, the sequence of A mating type loci was obtained from the genome of the monokaryon APP7 by homologous sequence alignment, and the physical location of mip was mapped. Primers were designed according to the transcriptome of APM2-16 and the related information of the monokaryon APP7 genome. The HD gene sequences in monokaryons APP7 and M2S16 were obtained by PCR amplification and sequencing. It was revealed that the two had no homology in the nucleotide sequences, and both encoded homodomain transcription factors through bioinformatics analysis. This study is helpful to spawn genetic improvement of A. cornea.
The function of laccase at different developmental stages of Hypsizygus marmoreus was clarified. Thirteen laccase gene sequences predicted by transcription sequencing of Hypsizygus marmoreus were analyzed and identified, and a phylogenetic tree was constructed. Laccase activity and gene expression levels in different growth and development stages were detected. The results showed that 10 of the 13 gene fragments were laccase genes. The evolutionary relationship between different laccase isoenzymes is obvious difference. Most laccases are closely related to those of wood rot fungi (Flammulina filiformis and Pleurotus spp.). Enzyme activity detection in different growth and development stages of Hypsizygus marmoreus showed that the laccase activity increased gradually from the mycelial regeneration stage to pinhead stage, while decreased gradually in the later stage of fruiting body formation. RT-qPCR of mycelial samples cultured in 40d, 60d, and 80d and samples in different growth and development stages was performed. The results revealed that during the period of vegetative growth of mycelium, the expression of most laccase genes continually increased by 1-3 times in 40-60d, and decreased in 60-80d. However, during reproductive period, the relative expression of most laccase genes reached the maximum level at pigmentation stage or primordium stage, but decreased at fruiting stage, consisting with the detection result of laccase activity. The relative expression of lcc3, lcc7, lcc8 and lcc9 increased by 10-100 times during the reproductive period of Hypsizygus marmoreus, indicating that different laccase might play different roles in the growth process from vegetative mycelium, interwoven mycelium to fruiting body formation and development, while highly expressed laccase genes might play a major role in matrix degradation and fruiting body formation.
The pileus of Volvariella volvacea easily opens after harvest and easily self-dissolves, oozily rots and smells in cold storage (below 10°C), making the fungus difficult to store. Previous studies showed that the Vvrin1 gene of MADS-box transcription factor might play a certain role in the elongation of stalk and the pileus expansion. In this study, the overexpression vector of Vvrin1 gene was constructed, and the heterokaryotic strain H1521 mycelium block was transformed by agrobacterium-mediated method. Through hyodamycin resistance plate screening, the hyodamycin resistance gene and Vvrin1 gene overexpression special fragment were amplified by PCR, and the Vvrin1 gene expression level in the transformants were analyzed by qRT-PCR, and finally 8 relatively reliable transformants were obtained. Preliminary study of the phenotype of the transformants indicated that in which seven transformants grew faster than the wild strain H1521, showing significant difference (P<0.05), and the hyphae of transformants were denser, with deeper color on colony surface. It could be inferred that MADS-box transcription factor Vvrin1 gene might participate in regulation of hypha growth and pigment synthesis and accumulation in V. volvacea. The results provide the strain materials and data support for further study on the regulation mechanism of mature senescence (especially pileus expansion) of V. volvacea.
Bioinformatics analysis was performed of the manganese peroxidase 1 (MnP1) gene of Lentinula edodes. The strain 18 and its mutagenic strain 18N44 of L. edodes were selected as experimental materials, and gene expression and enzyme activity of manganese peroxidase during high temperature stress were determined. The results showed that LeMnP1 is located outside the cell. This protein belongs to the peroxidase superfamily and has high homology with that of basidiomycetes. The recovery ability of 18N44 was obviously higher than that of strain 18 after high temperature stress. During the high temperature treatment, the level of LeMnP1 gene expression in 18N44 was higher than that in 18, showing a tendency of first increasing and then decreasing. The MnP activity of 18N44 was significantly higher than that in 18 at 0h. With the prolongation of the duration of high temperature stress, MnP activity in 18N44 decreased significantly. However, LeMnP1 gene expression and enzyme activity remained stable in 18 during high temperature treatment. It is speculated that higher expression of LeMnP1 in 18N44 may be closely related to the faster recovery properties of 18N44 after high temperature stress.
Flammulina filiformis is a kind of low-temperature fruiting fungus. Its primordium formation and the development of fruiting bodies both require low temperature and the latter needs lower. Therefore, the process of factory production consumes large energy and is disadvantageous to industrialized low-cost production. In this study, transcriptome sequencing technology (RNA-Seq) was used to analyze F. filiformis mycelium development and primordium formation. A total of 7 935 differentially expressed genes was screened, of which 4 025 genes were up-regulated and 3 910 genes were down-regulated after primordium formation. Through analyzing the metabolic pathway bioinformatics such as GO annotations and KEGG pathway annotations, there is an inference from the metabolic regulation of cold-induced primordium formation. When the F. filiformis mycelium cells were in cold condition, the expression of genes related to sugar transporter decreased, leading to a decline in the efficiency of carbon source uptake, therefore the most of gene expression related to the glycolysis pathway were down-regulated, impelling a decrease of synthesis of acetyl CoA, the substrate of tricarboxylic acid cycle, and finally causing decrease in the cell energy output. This negative feedback signal up-regulated the genes expression of oxidative metabolism of lipids stored in cells, and produced acetyl CoA for the production capacity of tricarboxylic acid cycle. This negative feedback also led to the up-regulation of unsaturated fatty acid synthesis gene expression to regulate cell fluidity. Most gene expression of the phospholipid and sphingolipid metabolism pathways were up-regulated, and the synthesis was increased, changing the cell membrane components, and then the cells remodeled into another state. Most of the gene expression related to DNA replication, RNA transcription, and protein synthesis were up-regulated, indicating that the cell was in the period of vigorous proliferation when the primordium formed. The results of this study revealed at molecular level that the primordium formation of F. filiformis accompanied the transformation of energy sources, the mutual regulation of various metabolic pathways, and the impact of related genes expression, being helpful to breeding of new F. filiformis strains tolerant to high-temperature and reducing energy consumption.
The nutritional characteristics and taste and flavor of some edible fungi, fruits and vegetables were scientifically evaluated and compared. Four common edible fungi Flammulina filiformis, Hypsizygus marmoreus, Lentinula edodes and Agaricus bisporus, and four common fruits and vegetables Malus domestica, Musa acuminata, Daucus carota and Solanum lycopersicum were used as test materials. The free amino acids were measured by an automatic amino acid analyzer, and the ratios of free amino acid components and taste characteristics of these eatables were systematically compared. Principal component and cluster analysis showed that the four edible fungi, and four fruits and vegetables were significantly divided into two categories. The total quantity of free amino acids in edible fungi was on an average more than three times that in fruits and vegetables. Each free amino acid in the four edible fungi was proportionally balanced. The percentage of essential amino acids/(essential amino acids + non-essential amino acids) of the four edible fungi was around 40%, and that of the essential amino acids/non-essential amino acids was close to or over 60%, approaching the ideal value. The taste amino acids, umami amino acids, sweetness amino acids, aromatic amino acids and bitterness amino acids, of the four edible fungi were more than twice those of the four fruits and vegetables. The most significant TAV values appeared in glutamic acid, phenylalanine, glycine, threonine, histidine, lysine, valine, arginine and alanine. These amino acids belonged to five major amino acid families, and their metabolic pathways were the same in edible fungi and fruits and vegetables, with the exception of lysine. The results showed that the tested edible fungi had higher quality amino acid ratios and richer flavor components than fruits and vegetables. This difference was overall reflected in the metabolism of many amino acids rather than individual amino acids.
During analysis of the meiotic recombination hotspots, we speculated that there might be a third mat-B sublocus presenting in the genome of Flammulina filiformis. The genomes of the homokaryotic strains ‘6-3’ and ‘6-21’ were annotated, and 8 paired homologous pheromone receptor genes and 3 paired homologous pheromone precursor genes were found, which constituted two mat-B subloci (α and β) those were 326kb apart. An extra pheromone precursor gene was found in the strain ‘6-21’, which was absent in ‘6-3’, constituting the third mat-B sublocus γ unlinked with α and β. Heterocaryon formed by mating ‘6-3’ with ‘6-21’ was cultivated for fruiting, and 142 single spore strains were isolated from the fruit-bodies. As a result, there were 8 genotypes of mat-B confirmed by compatibility tests. The third sublocus of mat-B was verified to have the same function as α and β loci.
Agrocybe aegerita is a delicious mushroom possessing very high economic value. The functional genomic research of this mushroom has been developed progressively with the completion of genome sequencing. The efficient transformation system is the key to technical foundation of gene function research. In this study, the PEG mediated method for genetic transformation of A. aegerita was carried out. A transformed DNA fragment was constructed by employing hygromycin resistance gene hph as selection marker and enhanced green fluorescent protein gene egfp as reporting gene. The sensitivity assessment showed that the hygromycin B concentration of 150μg/mL could inhibit the mycelial growth of A. aegerita completely. The highest yield of protoplasts could be obtained by using 2% lysing enzyme to digest the fresh mycelia at 30°C for 3h. The constructed DNA fragments were transferred into the protoplasts by PEG-mediated method, and transformants were obtained with a transformation efficiency of 7 protoplasts/μg DNA through hygromycin resistance selection. Transformants were verified by PCR analysis and fluorescence microscopy observation. The results indicated that hph and egfp were successfully transformed into the protoplast, and they stably expressed in the strain of A. aegerita. The stable PEG‐mediated transformation system provided an essential technical basis of the study on gene function of this fungus.
The popular edible and medicinal fungus, shiitake (Lentinula edodes) has the highest yield among edible fungi in China, but it is often faced with the problem of degradation of cultivar characters. Di-mon cross breeding between the monokaryotic strain ‘808’ and other six strains (three wild and three cultivated strains) was carried out to screen the main cultivars suitable for local cultivation. Among generated 270 di-mon combinations, 89 hybrids were identified through observation of clamp connections and antagonistic test, of which 10 with qualified characteristics were selected via cultivation experiments. Comparison of the agronomic traits resulted in obtainment of excellent hybrids ZJXG 5 and ZJXG 8 having characteristics of exceptional segregation, good appearance quality, light brown color and high biological efficiency of 86%-88% in the first four refruiting period. Both were the off-springs of bikaryotic parents originated from the local wild strain S1 in Changbai Mountains and the local main cultivated strain BY1. This study revealed that the compatibility, fruiting rate and yield performance of the di-mon hybrids exhibited higher level when collection of the parent strains and selectivity of filial generations were conducted in the close region. Taking advantage of local wild L. edodes resources for breeding usually yields good cultivars adaptable to local natural environment.
MAT-type genes, ISSR polymorphism and genetic clustering of 166 strains originated from different hyphae sprouted from different spores and different germ pores of four wild Morchella spp. were analyzed and hybrid experiments were conducted. The results showed morphological and genetic differences are apparent among strains. Four ISSR polymorphic primers were selected from 12 primers for genetic diversity analysis and 22 polymorphic bands were amplified with average polymorphism ratio of 88.0%. Based on GS and UPGMA, the hyphae of 166 strains can be divided into two groups separately corresponding to Morchella sextelata (KIB1811) and M. importuna (KIB1754, KIB1760 and KIB7042). The cluster analysis suggested that strains from different hyphae sprouted from the same spore have genetic differences. The genetic differentiation of the hyphae of the different single-spores from the same fruiting body is evident while that from different fruiting bodies is more evident. The results of MAT-type genes’ test showed that strain from single-spore has only one MAT-type gene (MAT 1-1-1 or MAT 1-2-1) and strains originated from hyphae sprouted from single germ pores in the same spore harbor the same MAT-type gene. This demonstrated morel is heterothallic fungus. Compared with M. sextelata, the genetic diversity of M. importuna is more plentiful and genetic background is more abundant. ISSR analysis also showed that the genetic relationship between morels could not be accurately measured simply according to the morphological characteristics of mycelium and sclerotium. These results provide theoretical reference for accurate breeding of high-quality morel strains.
Pleurotus eryngii is a typical basidiomycete with a tetrapolar mating system. Usually, a pair of compatible monokaryotic mycelia is required to form a stable dikaryotic mycelium. In this study, the A and B mating genes (HD1 and HD2) and pheromone precursor genes PEphb3.1 and PEphb3.3 from P. eryngii monokaryotic strain 181 (A1B1) were cloned and transformed into the compatible monokaryotic strain 183 (A2B2) by PEG-mediated transformation, and 22 positive transformants were obtained. Eleven transformants with clamp connection were further observed in detail, and the clamps and nuclear phases were described. This study provides a methodological reference for further research on genetic manipulation of mating genes. The concept and application value of homonuclear dikaryon are discussed.
Typical laccases usually belong to the subfamily 1 of auxiliary activity family 1 (the AA1_1 family), while the multicopper oxidases in AA1_2 family usually possess an activity to oxidize Fe 2+ to Fe 3+, and some of the known AA1_2 enzymes simultaneously possess a laccase activity. The whole genome of Morchella importuna only possess a single gene of AA1_2 family. Whether the enzymatic protein encoded by this AA1_2 gene possesses a laccase function remains undetermined. This study is aiming at understanding the functions of the gene from its expression profiling in combination with biochemical characterization of the encoding enzyme. Gene expression levels at different stages during the growth and development of M. importuna were determined by quantitative real-time PCR. The protein-coding sequence of the gene was cloned into a pET vector and overexpressed in Escherichia coli cells. Purified enzymatic protein was obtained by chromatography and then determined for its biochemical characteristics. The results showed that the AA1_2 family multicopper oxidase gene had low expression levels in the vegetative mycelia presented in exogenous nutrient bags and the soils, and high expression levels in primordia and fruiting bodies. The purified enzymatic protein had a molecular weight near 64kDa, showing bifunctional activities of ferroxidase (EC 1.16.3.1) and laccase (EC 1.10.3.2). The ferroxidase activity showed the maximum level at pH 4, while the laccase activity showed the maximum level at pH 6. Both enzymatic activities were optimum at 30°C, and retained over 70% of the activities after incubation for 16h at 30°C. Both the ferroxidase and laccase activities could be inhibited by Mn 2+, Pb 2+ and Hg 2+. The enzymatic protein showed a high tolerance to the protein denaturants SDS and urea. The findings confirmed that the enzymatic protein encoded by the AA1_2 family multicopper oxidase gene possessed bifunctional ferroxidase-laccase activities with enzymological evidences, which is the first report in macrofungi of Ascomycota, and might provide insights into further studies on the involvement of iron metabolism as well as laccase activity in the formation and development of morel fruiting body.
Stropharia rugosoannulata is an important edible fungus rich in nutrition and displays high capacity of lignocellulose degradation. It is a heterothallic basidiomycete with a tetrapolar mating system. However, the structures of mating loci have not been characterized. In this study, mating type A and B loci were analyzed by bioinformatics based on genomic data. The results revealed that A locus was composed of conserved HD1 and HD2 genes and the synteny of several genes downstream and upstream of A loci was higher as compared with other fungi, including conserved MIP, Sec61 protein, glycine dehydrogenase and β-flanking protein. B locus consisted of 5 pheromone receptors and 3 pheromone precursor genes. Compared with other fungi, the synteny of B loci was poor, indicating that B loci were highly variable in different fungi. The results obtained would help us to uncover the mating locus structures of S. rugosoannulata, being helpful to genetic breeding of the fungus.
Laccase plays a primary role in the lignin-degrading ability of Lentinula edodes and exerts significant influence on the quantity and qualities of products. To elucidate comprehensive transcriptional profile in the high laccase-producing monokaryotic mycelium, the transcriptome sequencing was performed and a total of 15 522 genes was annotated from the two levels of laccase activity of L. edodes strains. GO (gene ontology) analysis indicated that differentially expressed genes (DGEs) were mass-enriched in oxidoreductase activity, including the genes related to lignin-degrading enzymes and cytochrome P450 genes. KEGG (Kyoto encyclopedia of genes and genomes) pathway analysis revealed that the DGEs were significantly enriched in starch and sucrose metabolism and pentose and glucuronate interconversions including the genes encoding glycoside hydrolase and UDPG dehydrogenase, which were specifically up-regulated. 172 DGEs encoding transcription factors identified by searching the database led to the prediction of bZIP, C2H2 and C4 subfamilies probably binding with the laccases genes. The transcriptome results suggested that the alteration of gene expression involved lignin degradation and carbohydrate metabolism, in particular the up regulation of genes involved in pentose and glucuronate pathway, resulting in the high-efficiency bioconversion of lignin-degrading products provided as nutrient to promote the L. edodes growth. Transcription factors might have a crucial role in the regulation of laccase activity. This study provides critical genetic data for further understanding the metabolic mechanism of the high-yielding laccase activity monokaryotic strain.
With the development of printing and dyeing industry in China, the harmfulness of wastewater to the ecological environment is becoming increasingly serious, and it is urgent to explore a degradation method with obvious decolorization and low cost. It was found that Auricularia cornea strain AC5 could degrade various types of dyes, especially triphenylmethane dye. Degradation experiment of different dyes (75.0mg/L) was carried out for 12h using the extracellular crude enzyme solution of A. cornea incubated in 7d at 26°C under the condition of shake culture (160r/min). The result showed that the degradation rates of triphenylmethane dye malachite green, crystal violet, anthraquinone dye reactive blue 19 and azo dye active blue 222 were 83.27%, 71.77%, 67.81% and 63.92%, respectively. Enzyme activity determination showed that the laccase activity reached up to 321.0U/mL when degradation rate of malachite green attained to the highest level. The activities of lignin peroxidase and manganese peroxidase were lower than that of laccase. It was inferred that laccase played a major role in the process of dyes’ degradation. This study proved that it was a low-cost and convenient method using the crude enzyme solution of A. cornea to degrade dye wastewater.
Tr2016, a new cultivar resulted from multiple domestication for years of a wild Tremella fuciformis strain isolated from rotten wood in Pillow Mountain of Youxi County, Fujian Province, was successfully bred. The hyphae have clamp connections, forming white or yellowish colonies. Fresh fruiting bodies are white, peony-shaped with average diameter of 13.2cm and height of 7.2cm, and consist of several wavy or crimped pieces, turning into light yellow when dry. Tr2016 yields fruiting bodies on an average of 114.0g per cultivated bottle when fresh and 34.1g when dry in 45 days after inoculation.
Hypsizygus marmoreus ‘Finc-B-7’ was bred by single-spore inbred cross breeding technique using Finc-B-3 strain as the parent. The fruiting bodies of new strain have very clear marble streaks. Antagonism tests prove that it is significantly different from the parent strain. The suitable culture temperature is 18-23°C, the optimal mycelial culture period is 70 days, and fruiting body formation period is 21 days. The average yield is 188g/bottle. It is a new high-yielding strain of H. marmoreus with good quality.
‘Lixiang-2’, a new cultivated strain of Lentinula edodes with thermotolerance, originated from a parental natural mutant strain ‘Wuxiang-1’, was selected by using multiple manual targeted selection technology. The process of systematic selection, evaluation and screening was conducted for 4 consecutive years. The fruit bodies of this strain have short stipe and firmly fleshy thick pileus with light color. This strain is characterized by early fruiting, obvious refruiting periodicity, and tardy opening, etc. In regional test, the average yield of each cultivated bag (bag size 15cm×55cm) reached 526g, being 15.2% higher than that of the starting strain. The mushroom quality is better than that of ‘Wuxiang-1’ and ‘L9319’, and this strain is suitable for cultivation in Zhejiang during summer and autumn.
‘Zhehuang 1#’, a new cultivated variety derived from domestic breeding of wild Sanghuangporus vaninii is reported. The sporophore is fan-shaped or horse’ hoof-shaped, 6.0-15.0×3.0-5.0cm, with thickness of 2.0-5.0cm at the base and 0.2-2.0cm at the edges, and surface colour ranged from gold to yellowish-brown. The context is yellowish-brown. The optimal temperature for mycelium growth is 25-30°C and for sporophore development 20-25°C. The sporophore contains total flavonoid, crude polysaccharide and total triterpene of 8.26%, 5.40% and 0.96%, respectively. ‘Zhehuang 1#’ is planted in spring and autumn in Zhejiang Province. It shows strong resistance to environmental stress, high rate of finished product, high grade commodification, and high yield (13.3g/bag), suitable for sack cultivation.
‘Shuangbao 106’ cultivar was isolated from a single-spore culture of Agaricus bisporus ‘A20’. Its sporophores are scattered, or rarely clustered, and approximately hemispherical. The cap is white, with smooth bright surface and dense texture, and 3.5-5.0cm in diameter and 2.0-3.0cm in thinkness. The stipe is 2.0-2.8cm long, stout, white, subcylindrical, and slightly inflated at the base. The optimal mycelial growth temperature is 22-25°C, and fruiting temperature is 10-24°C with optimum of 16-18°C. Fresh fruiting body contains 1.00% crude fibre, 3.54% protein and 2.84% total amino acids. ‘Shuangbao 106’ is characterised by its early fruiting, good appearance of sporophore, high yield, and high grade commodification. It is suitable for industrialized cultivation.
