Study on the Function of ABC Transporter Gene LeABC2 in Thermotolerance of Lentinula edodes

doi:10.16420/j.issn.0513-353x.2025-0545

Acta Horticulturae Sinica ›› 2026, Vol. 53 ›› Issue (5): 1315-1328.doi: 10.16420/j.issn.0513-353x.2025-0545

• Genetic & Breeding · Germplasm Resources · Molecular Biology • Previous Articles Next Articles

Study on the Function of ABC Transporter Gene LeABC2 in Thermotolerance of Lentinula edodes

WU Yulong¹, WANG Yifei¹, LIU Gaolei¹^,², WANG Lu¹, ZHAO Zhidong¹, WANG Zhuoren³, XIAO Yang¹^,^*()

¹ Institute of Applied Mycology/State Key Laboratory of Agricultural Microbiology/Hubei Hongshan Laboratory， Huazhong Agricultural University， Wuhan 430070， China
² Institute of Vegetable， Wuhan Academy of Agricultural Sciences and Technology， Wuhan 430065， China
³ Institute of Economic Crops， Hubei Academy of Agricultural Sciences， Wuhan 430064， China

Received:2025-11-13 Revised:2025-12-01 Online:2026-05-25 Published:2026-05-26
Contact: XIAO Yang

Abstract

Abstract:

In this study，thermotolerance phenotypes of 126 Lentinula edodes strains were evaluated. Through genome-wide association analysis，genetic loci associated with thermotolerance in L. edodes were identified. Further investigation revealed a significant correlation between the ABC transporter gene LeABC2 and thermotolerance in L. edodes. Upon silencing the LeABC2 gene via RNAi，the recovery growth rates of the transformant strains after high-temperature stress were significantly enhanced，and the critical high-temperature thresholds were higher compared to the wild-type strain. These results indicated that the LeABC2 gene negatively regulates the mycelial thermotolerance of L. edodes.

Key words: Lentinula edodes, high temperature stress, thermotolerance, ABC transporter, genome-wide association study

WU Yulong, WANG Yifei, LIU Gaolei, WANG Lu, ZHAO Zhidong, WANG Zhuoren, XIAO Yang. Study on the Function of ABC Transporter Gene LeABC2 in Thermotolerance of Lentinula edodes[J]. Acta Horticulturae Sinica, 2026, 53(5): 1315-1328.

Figures/Tables 11

Fig. 1 Method for determining the thermotolerance coefficient of Lentinula edodes

Table 1 Primer sequences used in this study

引物名称 Primer name	引物序列（5′-3′） Primer sequence
ABC-DNA-F	TGGCTGTGGACAGTCTCTACAGTAT
ABC-DNA-R	GTCTAATACTACTTTAGAAC
ABC-cDNA-F	TGGCTGTGGACAGTCTCTACA
ABC-cDNA-R	GTCTAATACTACTTTAGAACG
ABC-RNAi-F	attgactgcttgaatggtaccCACACGAGGGCTCAGTCGC
ABC-RNAi-R	ctcgcagctcttcacgaattcGCAGTTTGCTGCAGACCCA
Leactin-F	ccacctcaaacttcggaattcGCAGTATTTATACCTACGGAGCGG
Leactin-R	tcttccgag CGTGAAGAGCTGCGAGTGTTG
Legpd-F	tgccactggcTGAAAAGACATGGATTGAGCCA
Legpd-R	tctagaggatccccgggTACCGTACATCCCTTGCTCTGC
qPCR-F	TGGTATTGGACCGTTGGATTT
qPCR-R	CGGCGAAAGAGACATAGTAGAAG
Leactin-qPCR-F	GCATCCTGTCCTTCTTACCGAG
Leactin-qPCR-R	AAGAGCGAAACCCTCGTAGATG
GPD-F	TTCGTTTCAGGCCTTGCCTC
HYG-R	CGGTGTCGTCCATCACAGTT

Fig. 2 Vector map of the recombinant plasmid LeABC2-RNAi

Table 2 Mycelial thermotolerance coefficients among the ten representative strains of Lentinula edodes under different high temperatures

菌株 Stain	33 ℃	35 ℃	37 ℃	39 ℃	41 ℃
香430 Xiang 430	0.791	0.782	0.951	0.772	—
日丰34 Rifeng 34	0.952	0.852	0.870	0.471	—
香9 Xiang 9	0.843	0.804	0.743	0.457	—
LEQC76	0.874	0.925	0.705	0.528	—
LMLH14	0.795	0.756	0.684	0.516	—
GAN059	0.602	0.796	0.783	0.580	—
LMLHL17	0.992	0.992	0.350	0.400	—
LMLH11	0.876	0.723	0.729	0.455	—
LEQC79	0.792	0.785	0.721	0.298	—
LPZH211	0.805	0.805	0.758	0.405	—
均值Mean	0.832	0.822	0.729	0.488	—
标准差 Standard deviation	0.101	0.077	0.148	0.120	—

Fig. 3 The frequency distribution of the thermotolerance coefficients of the 126 Lentinula edodes strains under 37 ℃ high temperature stress

Fig. 4 Genome-wide association study of the thermotolerance traits in the 126 Lentinula edodes strains A：Manhattan plot of SNPs associated with thermotolerance. The black horizontal dashed line indicates the genome-wide significance threshold（P = 9.03 × 10-7，Zhang et al.，2002）used to screen for loci strongly associated with the trait. The arrow points to a SNP on Scaffold_34（position：34：208168）whose value significantly exceeded the genome-wide threshold，suggesting a strong association with thermotolerance. B：Quantile-quantile（Q-Q）plot. The diagonal dashed line represents the expected distribution of observed values if no SNPs were associated with the trait. The deviation of observed points from this line indicates potential trait-associated SNPs

Fig. 5 Gene structure of LeABC2（A）and phylogenetic tree of the ABC2 protein（B）

Fig. 6 Vector map and identification of the recombinant plasmid LeABC2-RNAi M1：BM2000 + DNA Marker；1-8：Positive RNAi transformants of E. coli

Fig. 7 PCR identification of the transformants A：PCR identification of positive Agrobacterium colonies（M1：BM2000+ DNA marker，1-12：Positive RNAi transformants of Agrobacterium）；B：Identification of the gene fragment from RNAi transformants with primers GPD-F and HYG-R（M1：BM 2000+ DNA marker；1-10：RNAi transformants RNAi-1 to RNAi-10；11：Plasmid LeABC2-RNAi used as positive control；12：DNA from S606 used as negative control；13：DNA from control used as negative control；14：Sterile water used as negative control）

Fig. 8 qRT-PCR determination of the relative expression level of LeABC2 in the transformants S606：Wild-type strain；Control：Control strain；R-1 and R-2：RNAi transformants ** P < 0.01

Fig. 9 Growth conditions of Lentinula edodes strains after heat stress under different temperatures ** P < 0.01，*** P < 0.001，**** P < 0.0001

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Study on the Function of ABC Transporter Gene LeABC2 in Thermotolerance of Lentinula edodes

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[14]	DENG Jiaoyan，HUANG Bin，Lü Lijun，WANG Jun，YU Xianchang，HE Chaoxing，YAN Yan，and LI Yansu*. Mechanisms of Foliar-spraying 1-MCP to Alleviate Injury of Pepper Seedlings Caused by High Temperature [J]. ACTA HORTICULTURAE SINICA, 2019, 46(5): 891-900.
[15]	KONG Lingjie，CHEN Yanbo，and WANG Yaqin*. Studies on Heat Tolerance of Different Summer Chrysanthemum Cultivars [J]. ACTA HORTICULTURAE SINICA, 2019, 46(12): 2437-2448.