等离子体诱变选育高产胞外多糖花脸香蘑菌株

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

摘要/Abstract

摘要：

花脸香蘑多糖具有较大的药用价值,选育高产多糖花脸香蘑菌株可丰富种质资源并促进花脸香蘑多糖科技创新和产业发展。以花脸香蘑原生质体单核体菌株LS01_10作为出发菌株,利用等离子体诱变技术选育高产胞外多糖的花脸香蘑菌株。在诱变功率为200 W,诱变时间为60 s的条件下,共获得5个性状稳定遗传的高产胞外多糖诱变菌株,其胞外多糖的产量是出发菌株的1.2-2.4倍。诱变菌株的菌丝形态和颜色都发生不同程度改变,且生物量均高于出发菌株。ISSR实验结果表明,诱变菌株与出发菌株相比在遗传水平发生不同程度的改变。高产多糖诱变菌株为进一步解析多糖高产机制提供了重要的菌种资源。

关键词: 花脸香蘑, 等离子体诱变, 胞外多糖

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

范敏, 王丽宁. 等离子体诱变选育高产胞外多糖花脸香蘑菌株[J]. 生物技术通报, 2021, 37(11): 119-124.

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

图/表 6

表1 供试ISSR引物及序列

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

图1 等离子体诱变效应曲线

Fig.1 Dosage-time curve of plasma mutagenesis

图2 不同碳源和氮源培养LS01_10胞外多糖含量 A:氮源为蛋白胨;B:碳源为麦芽糖;不同小写字母代表差异显著（P<0.05）,下同

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

图3 诱变菌株的胞外多糖含量

Fig.3 Exopolysaccharide contents of mutant strains

图4 出发菌株和突变菌株液、固形态（A）及生物量（B）

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

图5 引物ISSR1、ISSR9和ISSR11的PCR扩增电泳图谱

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

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