Development of Novel Polymorphic EST-SSR Markers in Bailinggu (Pleurotus tuoliensis) for Crossbreeding

Abstract

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.

Genome Sequencing Illustrates the Genetic Basis of the Pharmacological Properties of Gloeostereum incarnatum

“…AAs were the next most abundant family of CAZymes in G. incarnatum ; AAs identified in this species included 21 AA9, 18 AA1, and 14 AA3 enzyme families. These three AA families are also the most abundant AAs in other fungi [10,11,12,13]. However, only three AA2 family proteins, the lignin-modifying fungal peroxidases (PODs), were identified in G. incarnatum .…”

“…With the rapid advancement of sequencing technologies, the number of available fungal genomes has increased [9,10]. However, genomes of medicinal mushrooms remain scarce.…”

Pleurotus eryngii Genomes Reveal Evolution and Adaptation to the Gobi Desert Environment

“…The protoplast-derived monokaryons of PEE81 from Europe and PEF12 from China were isolated according to Dai et al (2017), which were used for the subsequent whole genome sequencing. Dikaryotic mycelia of PEE81 and PEF12 were cultured in a liquid Malt Yeast Extract Glucose (MYG) medium for 7 and 10 days (d), respectively.…”

Genomic Analyses Provide Insights Into the Evolutionary History and Genetic Diversity of Auricularia Species

“…De novo genome sequencing was conducted on the monokaryon A. cornea strain AC1 (hereafter, AC1) that was derived from the cultivated strain CCMJ2567 using protoplast monokaryogenesis technique (Dai et al, 2017), but with a lywallzyme incubation time of 5 h. A total of 24 Wood Ear strains were used for resequenced the whole genome, including 12 A. cornea strains (seven cultivated strains from China and five wild individuals from Africa) and 12 A. heimuer strains from China (six cultivated strains and six wild individuals) (Supplementary Table S1). These strains were cultured on Malt Yeast Extract Glucose (MYG) medium (10 g/L maltose, 5 g/L yeast extract powder, and 5 g/L glucose) with cellophane sheets and then incubated at 24°C for 9 days.…”

“…PCRs were then conducted in reaction conditions and thermal cycling protocols as described previously (Fu et al, 2017), but with an annealing temperature of 59°C. PCR products were confirmed using silver staining (Dai et al, 2017).…”