干扰苯丙氨酸解氨酶基因pal1延迟糙皮侧耳发育的分子机制

doi:10.1016/j.jia.2024.06.002

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (4): 1477-1488.DOI: 10.1016/j.jia.2024.06.002

干扰苯丙氨酸解氨酶基因pal1延迟糙皮侧耳发育的分子机制

收稿日期:2023-11-16 接受日期:2024-05-23 出版日期:2025-04-20 发布日期:2025-03-17

Molecular mechanism of delayed development by interfering RNA targeting the phenylalanine ammonia lyase gene (pal1) in Pleurotus ostreatus

Qi He^{1, 2}, Yuqing Jiang², Chenyang Huang², Lijiao Zhang², Ludan Hou^{4, 5}, Fangjie Yao^{1, 3#}, Mengran Zhao^2#

¹Engineering Research Center of Edible and Medicinal Fungi, Ministry of Education/Jilin Agricultural University, Changchun 130118, China
²State Key Laboratory of Efficient Utilization of Arable Land in China/Key Laboratory of Microbial Resources Collection and Preservation of Ministry of Agriculture and Rural Affairs, Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China
³College of Horticulture, Jilin Agricultural University, Changchun 130118, China
⁴College of Food Science and Engineering, Shanxi Agricultural University, Taigu 030801, China
⁵Shanxi Key Laboratory of Edible Fungi for Loess Plateau, Taigu 030801, China

Received:2023-11-16 Accepted:2024-05-23 Online:2025-04-20 Published:2025-03-17
About author:Qi He, E-mail: heqi_jlau@126.com; #Correspondence Fangjie Yao, E-mail: yaofj@aliyun.com; Mengran Zhao, E-mail: zhaomengran@caas.cn
Supported by:
This study was supported by the National Key R&D Program of China (2022YFD1200600), the National Natural Science Foundation of China (32002110) and the earmarked fund for China Agriculture Research System (CARS-20).

摘要/Abstract

摘要：

食用菌发育受阻会影响子实体的生产周期和产量。苯丙氨酸解氨酶（PAL，EC 4.3.1.5）是一种催化苯丙氨酸脱氨生成反式肉桂酸的酶。前期研究结果发现，糙皮侧耳pal1基因的转录降低能够延缓子实体发育。因此，我们以野生型（WT）和RNA干扰（RNAi）菌株为材料，利用转录组测序和农杆菌介导的遗传转化方法，研究了pal1基因的分子调控机制。结果表明，干扰pal1基因导致PAL酶活性和总酚含量下降胞内H₂O₂含量增加。RNA-Seq数据表明，KEGG通路显著富集在过氧化物酶体途径、MAPK信号途径-酵母和另外三条途径，编码过氧化氢酶的基因cat1参与了上述显著富集的多个通路。外源H₂O₂能够显著增强cat1基因的转录，提高CAT总酶活性。添加H₂O₂清除剂后，RNAi-pal1菌株的cat1基因转录水平和CAT酶活性显著高于野生型菌株，表明pal1通过影响胞内的H₂O₂含量来调节cat1的表达。过表达糙皮侧耳的cat1基因导致了生长迟缓，尤其是在原基形成过程中。综上所述，本研究阐明了PAL1通过信号分子H₂O₂影响cat1基因的表达从而调控了糙皮侧耳的发育。研究结果深化了对食用菌分子发育机制的理解。

Abstract:

Blocking the development of edible mushrooms will affect the production cycle and yield of fruiting bodies. Phenylalanine ammonia lyase (PAL, EC 4.3.1.24.) is an enzyme that catalyzes the deamination of phenylalanine to form trans-cinnamic acid. Previous studies have shown that a decrease in pal1 gene transcription delays fruiting body development in Pleurotus ostreatus. Herein, we used wild type (WT) and RNA interference (RNAi) strains to study the molecular regulation of pal1 by RNA sequencing and Agrobacterium-mediated genetic transformation. Our results showed that interference with the pal1 gene resulted in reductions in the total PAL enzyme activity and the total phenol content, as well as an increase in the intracellular H₂O₂ content. RNA-Seq data demonstrated that the significantly enriched KEGG terms were mainly related to the peroxisome pathway, MAPK signaling pathway-yeast and three other pathways, and the catalase (CAT) gene cat1 is also involved in multiple pathways that were enriched above. Exogenous H₂O₂ significantly enhanced the transcription of the cat1 gene and elevated total CAT enzymatic activity. Moreover, the levels of cat1 gene transcription and the total CAT enzymatic activity in the RNAi-pal1 strains gradually become closer to those in the WT strain through the removal of H₂O₂, which indicated that pal1 regulated the expression of cat1 by affecting the intracellular H₂O₂ content. Finally, the overexpression of the cat1 gene in P. ostreatus caused growth retardation, especially during the process of primordia formation. In conclusion, this study demonstrated that PAL1 affects cat1 gene expression through the signaling molecule H₂O₂ and regulates the development of P. ostreatus. The findings of this study enhance our understanding of the molecular developmental mechanism of edible mushrooms.

Key words: Pleurotus ostreatus , phenylalanine ammonia lyase , catalase , development , regulation

Qi He, Yuqing Jiang, Chenyang Huang, Lijiao Zhang, Ludan Hou, Fangjie Yao, Mengran Zhao. 干扰苯丙氨酸解氨酶基因pal1延迟糙皮侧耳发育的分子机制[J]. Journal of Integrative Agriculture, 2025, 24(4): 1477-1488.

Qi He, Yuqing Jiang, Chenyang Huang, Lijiao Zhang, Ludan Hou, Fangjie Yao, Mengran Zhao. Molecular mechanism of delayed development by interfering RNA targeting the phenylalanine ammonia lyase gene (pal1) in Pleurotus ostreatus[J]. Journal of Integrative Agriculture, 2025, 24(4): 1477-1488.

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干扰苯丙氨酸解氨酶基因pal1延迟糙皮侧耳发育的分子机制

Molecular mechanism of delayed development by interfering RNA targeting the phenylalanine ammonia lyase gene (pal1) in Pleurotus ostreatus

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