Preliminary Study on the Mechanism of Signal Molecule H2O2 Regulating the Formation of Pleurotus ostreatus Cap Color

doi:10.16420/j.issn.0513-353x.2022-0546

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (6): 1243-1254.doi: 10.16420/j.issn.0513-353x.2022-0546

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

Preliminary Study on the Mechanism of Signal Molecule H₂O₂ Regulating the Formation of Pleurotus ostreatus Cap Color

HOU Ludan¹^,³, YAN Kexing¹, DONG Shuai¹, LIU Xingyu¹, CHANG Mingchang¹^,², MENG Junlong¹^,²^,^*()

¹College of Food Science and Engineering，Shanxi Agricultural University，Taigu，Shanxi 030801，China
²Shanxi Engineering Research Center of Edible Fungi，Taigu，Shanxi 030801，China
³Shanxi Key Laboratory of Edible Fungi for Loess Plateau，Taigu，Shanxi 030801，China

Received:2022-12-15 Revised:2023-04-21 Online:2023-06-25 Published:2023-06-27
Contact: * （E-mail：mengjunlongseth@hotmail.com）

Abstract

Abstract:

By spraying H₂O₂ and its scavenger N，N'-dimethylthiourea（DMTU）externally，it was clarified that H₂O₂ can participate in regulating the formation of Pleurotus ostreatus cap color. Using the reported Agaricus bisporus as a reference，28 genes involved in the melanin synthesis pathways were identified through sequence alignment，and the effect of exogenous H₂O₂ on the relative expression of key enzyme genes in different pathways was analyzed by qPCR. The results showed that H₂O₂ signaling molecule can change the proportion of different types of melanin in P. ostreatus and affecting the color of the cap by regulating the expression of key enzyme genes in different melanin synthesis pathways. On the other hand，it can promote the high expression of copper transcription factor genes mac，copper transporter genes ctr and polyphenol oxidase gene ppo1，and finally affect the formation of cap color.

Key words: Pleurotus ostreatus, cap color, hydrogen peroxide, polyphenol oxidase, copper ion transport

CLC Number:

S646.141

HOU Ludan, YAN Kexing, DONG Shuai, LIU Xingyu, CHANG Mingchang, MENG Junlong. Preliminary Study on the Mechanism of Signal Molecule H₂O₂ Regulating the Formation of Pleurotus ostreatus Cap Color[J]. Acta Horticulturae Sinica, 2023, 50(6): 1243-1254.

Figures/Tables 9

References 39

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引物	正向序列（5′-3′）	反向序列（5′-3′）
Primer	Forward sequence	Reverse sequence
qPCR_CM	GGGTAAATGGAGACGCAGAG	CTGGATTGGCTTGGAATTTGG
qPCR_4ABS	TGGGACGTACTCAGCAAAAG	ACACCAGTAACCAAAGACCC
qPCR_4ABH_1	CTCCCTACCTTCAGTGCATTC	TGAGGTGCTTGTTCTTTCCC
qPCR_4ABH_2	CTCCGAACATGACCCGAATAC	CTCACTAATCCCAAAGGCTCTC
qPCR_4ABH_3	GTTGTAGACTGTGAGAGGGAAG	TGAGACAAAGAATGGGAGTGG
qPCR_4ABH_4	GCAAGTCACATTCAAGGTCATC	TCCAGATTGCTATCGCTTCG
qPCR_GGT_1	CTGTCGAAAAGTTTGGTAGCG	CGTTGGGAGATGCATTTTGG
qPCR_GGT_2	GGTGGAGAACTATGACTTTCGG	TGCCGTGTCGTTCATATAAGC
qPCR_PDH	GTTTGCGACATCCCTGAAAAG	CGATGAACTCCGCTTCTACTG
qPCR_AT_1	CCTGATGCTTTCTACACCTCC	GTGAATGGAAAGGTTGTCGC
qPCR_AT_2	GGAGCGGTTGTGATATCTATAGG	TGAGGACATGAAGGATGGTTG
qPCR_ppo1	ACCCTTTGTTCTGGCTTCATC	TTCGTGATATTGTCCAGGCG
qPCR_ppo2	ACCCCAACCTTCAAAGTACAC	TGAGATCCATTGACTGCCAC
qPCR_ppo3	AAAGTGGGCTCCTAACGATC	GTAATGTTCTCGAAACGCTGG
qPCR_ppo4	GTCCAACGATCCCATTTTCTG	ATGAGCGGGAAAGGTATGAAG
qPCR_mac1	CAAAACAAAGGTGCCAGACAG	CATTCACCGCCAGTTTTACAC
qPCR_mac2	CCTATTGAACCAGCTCTTCCC	CATTGGCATCCACGAGTACT
qPCR_ctr1	TTTGCCATACAGTCCCTTCTC	CGATGAGATAGCCAGCATGG
qPCR_ctr2	TTACTCGCTGCCTTTCGG	ATCCTTTTCTCCCATCGACG
qPCR_ctr3	AACCACATCTTCGACTACCAC	AACCACATCTTCGACTACCAC
qPCR_ctr4	GGTTTTCATGACATACAACGCG	TGGGAGTTGAAGACGAAATGG
tubulin	AGGCTTTCTTGCATTGGTACACGC	TATTCGCCTTCTTCCTCATCGGCA

引物	正向序列（5′-3′）	反向序列（5′-3′）
Primer	Forward sequence	Reverse sequence
qPCR_CM	GGGTAAATGGAGACGCAGAG	CTGGATTGGCTTGGAATTTGG
qPCR_4ABS	TGGGACGTACTCAGCAAAAG	ACACCAGTAACCAAAGACCC
qPCR_4ABH_1	CTCCCTACCTTCAGTGCATTC	TGAGGTGCTTGTTCTTTCCC
qPCR_4ABH_2	CTCCGAACATGACCCGAATAC	CTCACTAATCCCAAAGGCTCTC
qPCR_4ABH_3	GTTGTAGACTGTGAGAGGGAAG	TGAGACAAAGAATGGGAGTGG
qPCR_4ABH_4	GCAAGTCACATTCAAGGTCATC	TCCAGATTGCTATCGCTTCG
qPCR_GGT_1	CTGTCGAAAAGTTTGGTAGCG	CGTTGGGAGATGCATTTTGG
qPCR_GGT_2	GGTGGAGAACTATGACTTTCGG	TGCCGTGTCGTTCATATAAGC
qPCR_PDH	GTTTGCGACATCCCTGAAAAG	CGATGAACTCCGCTTCTACTG
qPCR_AT_1	CCTGATGCTTTCTACACCTCC	GTGAATGGAAAGGTTGTCGC
qPCR_AT_2	GGAGCGGTTGTGATATCTATAGG	TGAGGACATGAAGGATGGTTG
qPCR_ppo1	ACCCTTTGTTCTGGCTTCATC	TTCGTGATATTGTCCAGGCG
qPCR_ppo2	ACCCCAACCTTCAAAGTACAC	TGAGATCCATTGACTGCCAC
qPCR_ppo3	AAAGTGGGCTCCTAACGATC	GTAATGTTCTCGAAACGCTGG
qPCR_ppo4	GTCCAACGATCCCATTTTCTG	ATGAGCGGGAAAGGTATGAAG
qPCR_mac1	CAAAACAAAGGTGCCAGACAG	CATTCACCGCCAGTTTTACAC
qPCR_mac2	CCTATTGAACCAGCTCTTCCC	CATTGGCATCCACGAGTACT
qPCR_ctr1	TTTGCCATACAGTCCCTTCTC	CGATGAGATAGCCAGCATGG
qPCR_ctr2	TTACTCGCTGCCTTTCGG	ATCCTTTTCTCCCATCGACG
qPCR_ctr3	AACCACATCTTCGACTACCAC	AACCACATCTTCGACTACCAC
qPCR_ctr4	GGTTTTCATGACATACAACGCG	TGGGAGTTGAAGACGAAATGG
tubulin	AGGCTTTCTTGCATTGGTACACGC	TATTCGCCTTCTTCCTCATCGGCA

关键酶编号 Key enzyme number	基因功能 Gene function	基因缩写 Gene abbreviation	基因编号 Gene number
Ⅰ	分支酸变位酶 Chorismate mutase	CM	g11035
Ⅱ	4-氨基苯甲酸合成酶 4-Aminobenzote synthase	4ABS	g1809
Ⅲ	4-氨基苯甲酸羟化酶1 4-Aminobenzote hydroxylase_1	4ABH_1	g2093
Ⅲ	4-氨基苯甲酸羟化酶2 4-Aminobenzote hydroxylase_2	4ABH_2	g3895
Ⅲ	4-氨基苯甲酸羟化酶3 4-Aminobenzote hydroxylase_3	4ABH_3	g12346
Ⅲ	4-氨基苯甲酸羟化酶4 4-Aminobenzote hydroxylase_4	4ABH_4	g9216
Ⅳ	γ-谷氨酰胺转移酶1 γ-Glutaminyltransferase_1	GGT_1	g649
Ⅳ	γ-谷氨酰胺转移酶2 γ-Glutaminyltransferase_2	GGT_2	g8358
Ⅴ	预苯甲酸脱水酶 Prephenate dehydratase	PDHtase	g9753
Ⅵ	预苯酸脱氢酶 Prephenate dehydrogenase	PDH	g10441
Ⅶ/Ⅷ	（4-羟基）苯基丙酮酸氨基转移酶1 (4-Hydroxy) phenylpyruvate aminoransferase_1	AT_1	g4186
Ⅶ/Ⅷ	（4-羟基）苯基丙酮酸氨基转移酶2 (4-Hydroxy) phenylpyruvate aminoransferase_2	AT_2	g7673
Ⅸ	多酚氧化酶1 Polyphenooxidase_1	PPO1	g7596
Ⅸ	多酚氧化酶2 Polyphenooxidase_2	PPO2	g3399
Ⅸ	多酚氧化酶3 Polyphenooxidase_3	PPO3	g3812
Ⅸ	多酚氧化酶4 Polyphenooxidase_4	PPO4	g11413
Ⅹ	苯丙氨酸解氨酶1 Phenylalnine ammonialyase_1	PAL1	g9868
Ⅹ	苯丙氨酸解氨酶2 Phenylalnine ammonialyase_2	PAL2	g9752
Ⅻ	4-香豆酸辅酶A连接酶1 4-Coumarate CoA ligase_1	4CL1	g5055
Ⅻ	4-香豆酸辅酶A连接酶2 4-Coumarate CoA ligase_2	4CL2	g1908
Ⅻ	4-香豆酸辅酶A连接酶3 4-Coumarate CoA ligase_3	4CL3	g7302
Ⅻ	4-香豆酸辅酶A连接酶4 4-Coumarate CoA ligase_4	4CL4	g8331
Ⅻ	4-香豆酸辅酶A连接酶5 4-Coumarate CoA ligase_5	4CL5	g11056
Ⅻ	4-香豆酸辅酶A连接酶6 4-Coumarate CoA ligase_6	4CL6	g1913
Ⅻ	4-香豆酸辅酶A连接酶7 4-Coumarate CoA ligase_7	4CL7	g5185
Ⅻ	4-香豆酸辅酶A连接酶8 4-Coumarate CoA ligase_8	4CL8	g333
Ⅻ	4-香豆酸辅酶A连接酶9 4-Coumarate CoA ligase_9	4CL9	g9792
Ⅻ	4-香豆酸辅酶A连接酶10 4-Coumarate CoA ligase_10	4CL10	g9146

关键酶编号 Key enzyme number	基因功能 Gene function	基因缩写 Gene abbreviation	基因编号 Gene number
Ⅰ	分支酸变位酶 Chorismate mutase	CM	g11035
Ⅱ	4-氨基苯甲酸合成酶 4-Aminobenzote synthase	4ABS	g1809
Ⅲ	4-氨基苯甲酸羟化酶1 4-Aminobenzote hydroxylase_1	4ABH_1	g2093
Ⅲ	4-氨基苯甲酸羟化酶2 4-Aminobenzote hydroxylase_2	4ABH_2	g3895
Ⅲ	4-氨基苯甲酸羟化酶3 4-Aminobenzote hydroxylase_3	4ABH_3	g12346
Ⅲ	4-氨基苯甲酸羟化酶4 4-Aminobenzote hydroxylase_4	4ABH_4	g9216
Ⅳ	γ-谷氨酰胺转移酶1 γ-Glutaminyltransferase_1	GGT_1	g649
Ⅳ	γ-谷氨酰胺转移酶2 γ-Glutaminyltransferase_2	GGT_2	g8358
Ⅴ	预苯甲酸脱水酶 Prephenate dehydratase	PDHtase	g9753
Ⅵ	预苯酸脱氢酶 Prephenate dehydrogenase	PDH	g10441
Ⅶ/Ⅷ	（4-羟基）苯基丙酮酸氨基转移酶1 (4-Hydroxy) phenylpyruvate aminoransferase_1	AT_1	g4186
Ⅶ/Ⅷ	（4-羟基）苯基丙酮酸氨基转移酶2 (4-Hydroxy) phenylpyruvate aminoransferase_2	AT_2	g7673
Ⅸ	多酚氧化酶1 Polyphenooxidase_1	PPO1	g7596
Ⅸ	多酚氧化酶2 Polyphenooxidase_2	PPO2	g3399
Ⅸ	多酚氧化酶3 Polyphenooxidase_3	PPO3	g3812
Ⅸ	多酚氧化酶4 Polyphenooxidase_4	PPO4	g11413
Ⅹ	苯丙氨酸解氨酶1 Phenylalnine ammonialyase_1	PAL1	g9868
Ⅹ	苯丙氨酸解氨酶2 Phenylalnine ammonialyase_2	PAL2	g9752
Ⅻ	4-香豆酸辅酶A连接酶1 4-Coumarate CoA ligase_1	4CL1	g5055
Ⅻ	4-香豆酸辅酶A连接酶2 4-Coumarate CoA ligase_2	4CL2	g1908
Ⅻ	4-香豆酸辅酶A连接酶3 4-Coumarate CoA ligase_3	4CL3	g7302
Ⅻ	4-香豆酸辅酶A连接酶4 4-Coumarate CoA ligase_4	4CL4	g8331
Ⅻ	4-香豆酸辅酶A连接酶5 4-Coumarate CoA ligase_5	4CL5	g11056
Ⅻ	4-香豆酸辅酶A连接酶6 4-Coumarate CoA ligase_6	4CL6	g1913
Ⅻ	4-香豆酸辅酶A连接酶7 4-Coumarate CoA ligase_7	4CL7	g5185
Ⅻ	4-香豆酸辅酶A连接酶8 4-Coumarate CoA ligase_8	4CL8	g333
Ⅻ	4-香豆酸辅酶A连接酶9 4-Coumarate CoA ligase_9	4CL9	g9792
Ⅻ	4-香豆酸辅酶A连接酶10 4-Coumarate CoA ligase_10	4CL10	g9146

Preliminary Study on the Mechanism of Signal Molecule H₂O₂ Regulating the Formation of Pleurotus ostreatus Cap Color

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