Breeding of Lepista sordida with High-yield Exopolysaccharide by Plasma Mutagenesis

doi:10.13560/j.cnki.biotech.bull.1985.2021-0278

Abstract

Abstract:

The polysaccharide of Lepista sordida owns huge medicinal value,and breeding L. sordida strain with high-yield polysaccharide could not only enrich the germplasm resources,but also promote the technological innovation and industrial development. To obtain L. sordida strains with high-yield exopolysaccharide,the monokaryon strain LS01_10 derived from protoplast was used as parental strain and plasma mutagenesis was conducted. When the optimal mutagenesis power was 200 W and inducing time was 60 s,5 mutant strains with high-yield exopolysaccharide and stable inheritance were obtained. The exopolysaccharide of these 5 mutant strains was 1.2-2.4 times higher than that of the parental strain. The morphology and color of hyphae of the mutant strains varied at different degree,the biomass was higher than that of the parental strain. The variations in genetic level among mutant and parental strains were confirmed by inter-simple sequence repeat（ISSR）analysis. The mutant strains with high-yield polysaccharide provide important germplasm resource for further resolving the mechanism of high polysaccharide yield.

Key words: Lepista sordida, plasma mutagenesis, exopolysaccharide

FAN Min, WANG Li-ning. Breeding of Lepista sordida with High-yield Exopolysaccharide by Plasma Mutagenesis[J]. Biotechnology Bulletin, 2021, 37(11): 119-124.

Figures/Tables 6

Table 1 Primers and their sequences of ISSR for this study

引物 Primer	序列 Sequence（5'-3'）	退火温度 Annealing temperature/℃
ISSR1	AGAGAGAGAGAGAGAGT	50
ISSR9	CTCCTCCTCCTCCTC	58
ISSR11	GGGTGGGGTGGGGTG	52
ISSR20	CACCACACACACACACA	55
ISSR30	ATGTATGTATGTATGA	55

Fig.1 Dosage-time curve of plasma mutagenesis

Fig.2 Exopolysaccharide contents of LS01_10 cultured in different carbon and nitrogen resources Tryptone was used as nitrogen source in figure A,and maltose was used as carbon source in figure B. Different lowercase letters indicate significant differences（P<0.05）,the same below

Fig.3 Exopolysaccharide contents of mutant strains

Fig.4 Mycelial morphology cultured in liquid media,PDA plates（A）,and biomass（B）of parental and mutant strains

Fig.5 Agarose electrophoresis patterns of PCR products amplified by primer ISSR1,ISSR9 and ISSR11

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