Using agro-residues in bioreactors to produce mushrooms is an important component of eco-agriculture. Here, we tested the cultivation of Lentinula edodes with corn cob (CC) and corn straw (CS), and then evaluated the resulting enzyme activities, agronomic traits, textures and nutrient compositions of mushrooms. Laccase (T1 formula, 3.26 g U⁻¹) and carboxymethyl cellulase (T3 formula, 1.01 g U⁻¹) activities were the highest at the time for a complete substrate colonization stage (TCSC), while acidic xylanase activity was the highest (CK formula, 4.05 g U⁻¹) in the mushroom block to color-turned (TMBCT) stage. The biological efficiency of growth on the T6 formula was 8.82% higher than growth on the CK formula, wherein the low C/N ratio of the substrate had an obvious negative effect on yield while the mass ratio of pileus (MRP) of fruiting bodies did not change with mixed substrates. No significant differences were observed in mineral composition for CK formulas, but corn crop waste (CCW) formulas exhibited more optimal nutritional contents. A formula containing more corn cob and sawdust (SD) (sum of at least 70%) as the substrate can produce fruiting bodies with good hardness. These results indicate that the use of corn cobs as the main ingredient, mixed with sawdust and corn straw to grow L. edodes provides a more efficient use of agro-residues for growth. Thus, mixed agro-residue formulas have exceptional advantages in texture, nutrition of fruiting bodies, and yields.

Corn stalks are a major source of agricultural waste in China that have the potential for more efficient utilisation. In this study, we designed substrate formulas with different proportions of corn stalks to cultivate Pholiota microspora. The substrate formula for P. microspora cultivation that could partially or completely replace sawdust with corn stalks was selected through the analysis of mycelial growth rates, fruiting body traits, yield, biological efficiency, nutrients, and mineral composition. Our results showed that the substrate formula T2 (38% wood chips and 38% corn stalks) resulted in the highest yield of 275.66 ± 2.87 g per bag, which was 6.60% higher than that of formula CK, and the highest biological efficiency of 90.75 ± 0.04%, which was 4.58% higher than that of CK, with no significant differences from CK in terms of fruiting body traits, nutrients, or mineral composition. The substrate formula T1 (19% corn stalks) led to mushroom yields with the highest mineral and amino acid contents and was thus more suitable for the cultivation of medicinal P. microspora. Therefore, substrates comprising a mixture of corn stalks and sawdust can be used as a novel, inexpensive, and high-yield alternative for the cultivation of P. microspora.

Outbreaks of wet bubble disease (WBD) caused by Mycogone perniciosa are increasing across the world and seriously affecting the yield of Agaricus bisporus. However, highly WBD-resistant strains are rare. Here, we tested 28 A. bisporus strains for WBD resistance by inoculating M. perniciosa spore suspension on casing soil, and assessed genetic diversity of these strains using 17 new simple sequence repeat (SSR) markers developed in this study. We found that 10 wild strains originating from the Tibetan Plateau in China were highly WBD-resistant strains, and 13 cultivated strains from six countries were highly susceptible strains. A total of 88 alleles were detected in these 28 strains, and the observed number of alleles per locus ranged from 2 to 8. Cluster and genetic structure analysis results revealed the wild resources from China have a relatively high level of genetic diversity and occur at low level of gene flow and introgression with cultivated strains. Moreover, the wild strains from China potentially have the consensus ancestral genotypes different from the cultivated strains and evolved independently. Therefore, the highly WBD-resistant wild strains from China and newly developed SSR markers could be used as novel sources for WBD-resistant breeding and quantitative trait locus (QTL) mapping of WBD-resistant gene of A. bisporus.

Grifola frondosa is an economically important edible and medicinal mushroom usually produced on substrate consisting of sawdust supplemented with wheat bran. Cultivation of G. frondosa on crop straw (corn cob, corn straw, rice straw, and soybean straw) as a substrate was optimized by using the D-optimum method of the simplex-lattice design, and the alternative of crop straw as a substitute for sawdust in the substrate composition was determined by the optimized model. The results showed that there was a significant positive correlation existing between the yield and corn cob. The growth cycle was negatively correlated with sawdust, corn cob and soybean straw, with sawdust significantly shortening the growth cycle of G. frondosa. The optimized high-yielding formula included 73.125% corn cob, 1.875% rice straw, 23% wheat bran and 2% light calcium carbonate (CaCO3) (C/N = 48.40). The average yield of the first flush was 134.72 ± 4.24 g/bag, which was increased by 39.97% compared with the control formula. The biological efficiency (BE) was 44.91 ± 1.41%, which was increased by 38.53% compared with the control. Based on the results of this study, corn cob can replace sawdust as one of the main cultivation substrates of G. frondosa.

Brown blotch disease (BBD) caused by Pseudomonas tolaasii is one of the most devastating diseases of Pleurotus spp. worldwide. Breeding for resistant strains is the most effective method for controlling BBD. To identify resistant germplasm for BBD management, 97 strains comprising 21 P. cf. floridanus, 20 P. ostreatus, and 56 P. pulmonarius were screened by two different methods; namely, inoculation of the pathogen on the mushroom pileus (IMP) and on the spawned substrate (IMSS) under controlled conditions. Out of the 97 strains screened, 22 P. pulmonarius, and four P. cf. floridanus were moderately resistant to BBD using the IMP method. Eleven P. pulmonarius, six P. cf. florida, and one P. ostreatus strains were highly resistant to BBD using the IMSS method. All of the 97 strains showed varying degrees of susceptibility using the IMP method, but eight strains were completely resistant using the IMSS method. Combining these two methods, five strains were highly resistant (four P. pulmonarius and one P. cf. floridanus) and 11 were moderately resistant (eight P. pulmonarius and three P. cf. floridanus). The resistance sources to P. tolaasii identified in P. pulmonarius and P. cf. floridanus could be used for further breeding of Pleurotus spp.

Identification of monokaryons and their mating types and discrimination of hybrid offspring are key steps for the crossbreeding of Pleurotus tuoliensis (Bailinggu). However, conventional crossbreeding methods are troublesome and time consuming. Using RNA-seq technology, we developed new expressed sequence tag-simple sequence repeat (EST-SSR) markers for Bailinggu to easily and rapidly identify monokaryons and their mating types, genetic diversity and hybrid offspring. We identified 1110 potential EST-based SSR loci from a newly-sequenced Bailinggu transcriptome and then randomly selected 100 EST-SSRs for further validation. Results showed that 39, 43 and 34 novel EST-SSR markers successfully identified monokaryons from their parent dikaryons, differentiated two different mating types and discriminated F₁ and F₂ hybrid offspring, respectively. Furthermore, a total of 86 alleles were detected in 37 monokaryons using 18 highly informative EST-SSRs. The observed number of alleles per locus ranged from three to seven. Cluster analysis revealed that these monokaryons have a relatively high level of genetic diversity. Transfer rates of the EST-SSRs in the monokaryons of closely-related species Pleurotus eryngii var. ferulae and Pleurotus ostreatus were 72% and 64%, respectively. Therefore, our study provides new SSR markers and an efficient method to enhance the crossbreeding of Bailinggu and closely-related species.