Mutagenesis and Domestication of Wild Pholiota nameko by High-voltage Corona Electric Field Protoplasts

doi:10.11924/j.issn.1000-6850.casb2023-0853

Abstract

Abstract:

The purpose of this study is to screen new varieties of Pholiota nameko with rapid growth rate. Pholiota nameko protoplasts were prepared by enzymatic hydrolysis using wild Pholiota nameko from Alshan of Inner Mongolia as materials, and the protoplasts were mutated by high-voltage corona electric field. The protoplasts were cultured by coating method on PDA medium in order to obtain high quality mutant strains. The experimental results show that under the conditions of voltage of 6kv and processing time of 10 s-10 min, the Pholiota nameko protoplasts mutagenesis mortality rate is 30.51%-100%; When the voltage is 7 kV and the treatment time is 10 s-5 min, the mutagenesis mortality rate is 39.55%-100%, and the daily growth rate of regenerated strains from mutagenic protoplasts is higher than that of the control group. Taking a voltage of 6 kV as an example, the growth rate of protoplast regenerated strains mutagenized by 10 s are 119% higher than the control group. Quantitative analysis of laccase activity revealed that the laccase activity of protoplast regenerated strains under 6 kV and 3 min mutation conditions increased by 8.79 times compared to the control group on the 6^th day, which indicates that high-voltage corona discharge mutagenesis of wild Pholiota nameko protoplasts can improve the growth rate and quality of the strain, enhance laccase activity, and obtain positive mutant strains, providing a new direction for further domestication of wild Pholiota nameko.

Key words: Pholiota nameko, protoplast, high-voltage corona electric field, growth rate, laccase

CLC Number:

S646

LIU Chaoxiong, WANG Haiyan, MA Fengli, XU Xingyu, LI Yajiao, SUN Guoqin, FENG Tingting, GUO Jiufeng. Mutagenesis and Domestication of Wild Pholiota nameko by High-voltage Corona Electric Field Protoplasts[J]. Chinese Agricultural Science Bulletin, 2026, 42(11): 65-73.

Figures/Tables 14

References 24

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编号	电压/kV	时间	再生菌落个数/个	再生率/%	致死率/%
CK	0	0	66.67	0.02220	0.00
1	6	10 s	46.33	0.01545	30.51
2	6	30 s	46.00	0.01535	31.00
3	6	1 min	39.33	0.01310	41.01
4	6	3 min	37.00	0.01235	44.50
5	6	5 min	27.00	0.00900	59.50
6	6	10 min	0.00	0.00000	100.00
7	7	10 s	40.30	0.01345	39.55
8	7	30 s	37.00	0.01235	44.50
9	7	1 min	29.00	0.00965	56.50
10	7	3 min	23.00	0.00765	65.50
11	7	5 min	0.00	0.00000	100.00
12	7	10 min	0.00	0.00000	100.00

编号	电压/kV	时间	再生菌落个数/个	再生率/%	致死率/%
CK	0	0	66.67	0.02220	0.00
1	6	10 s	46.33	0.01545	30.51
2	6	30 s	46.00	0.01535	31.00
3	6	1 min	39.33	0.01310	41.01
4	6	3 min	37.00	0.01235	44.50
5	6	5 min	27.00	0.00900	59.50
6	6	10 min	0.00	0.00000	100.00
7	7	10 s	40.30	0.01345	39.55
8	7	30 s	37.00	0.01235	44.50
9	7	1 min	29.00	0.00965	56.50
10	7	3 min	23.00	0.00765	65.50
11	7	5 min	0.00	0.00000	100.00
12	7	10 min	0.00	0.00000	100.00

差异源	SS	df	MS	F	P-value	F crit
时间	1264.07	1	1264.07	16.70661	0.006448	5.987378
电场强度	4041.96	6	673.6599	8.90344	0.008799	4.283866
误差	453.9773	6	75.66288
总计	5760.007	13

差异源	SS	df	MS	F	P-value	F crit
时间	1264.07	1	1264.07	16.70661	0.006448	5.987378
电场强度	4041.96	6	673.6599	8.90344	0.008799	4.283866
误差	453.9773	6	75.66288
总计	5760.007	13

编号	电压/kV	时间	菌丝生长直径/mm
编号	电压/kV	时间	第5天	第6天	第7天	第8天
CK	0	0	17.03	18.02	21.73	24.47
1	6	10 s	21.60	22.67	28.67	37.87
2	6	30 s	17.07	20.33	24.50	29.70
3	6	1 min	14.83	17.17	19.43	22.05
4	6	3 min	22.77	22.00	26.67	32.27
5	6	5 min	20.40	25.23	27.33	32.13
7	7	10 s	14.17	16.90	21.50	25.53
8	7	30 s	19.57	23.83	26.13	29.10
9	7	1 min	16.80	18.90	28.30	33.33
10	7	3 min	18.03	20.57	23.97	27.33