The Molecular Mechanism of Phenylalanine Ammonia-Lyase Gene pal2 Regulating Pleurotus ostreatus Mycelial Growth Under Stimulation of Low-temperature and Light

doi:10.16420/j.issn.0513-353x.2022-0001

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (4): 831-841.doi: 10.16420/j.issn.0513-353x.2022-0001

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The Molecular Mechanism of Phenylalanine Ammonia-Lyase Gene pal2 Regulating Pleurotus ostreatus Mycelial Growth Under Stimulation of Low-temperature and Light

JIANG Yuqing¹, HE Qi¹^,², HOU Ludan³, WU Xiangli¹, HUANG Chenyang¹, and ZHAO Mengran¹^,^*()

1. Key Laboratory of Microbial Resources Collection and Preservation，Ministry of Agriculture and Rural Affairs，Institute of Agricultural Resources and Regional Planning，Chinese Academy of Agricultural Sciences，Beijing 100081，China
2. Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi，Jilin Agricultural University，Jilin，Changchun 130118，China
3. Key Laboratory of Shanxi Province for Loess Plateau Edible Fungi，Collaborative Innovation Center of Quality and Efficiency of Loess Plateau Edible Fungi in Shanxi，College of Food Science and Engineering，Shanxi Agricultural University，Jinzhong，Shanxi 030801，China

Received:2022-09-09 Revised:2022-12-26 Online:2023-04-25 Published:2023-04-27
Contact: *（E-mail：zhaomengran@caas.cn）

Abstract

Abstract:

Wild-type strain and pal2 gene interference strains of Pleurotus ostreatus were used as materials in this study. All the strains were incubated at 28 ℃ in the darkness for three days and then were treated with environmental stimulation（15 ℃，12 h light/12 h darkness）for six days. Strains incubated at 28 ℃ in the darkness without the former environmental stimulation were taken as control. The pal2 gene transcription level，enzyme activity，contents of enzyme substrate and product of all the strains treated with different environmental conditions were tested and compared. The effects of various concentrations of exogenous substrates and products on the mycelial phenotype were examined. The results preliminarily explained the reasons for the slowdown of mycelial growth under environmental stimulation. The results revealed that both phenylalanine ammonia-lyase and the gene pal2 responded to environmental stimulation，but there was an obivous response lag in enzyme activity，product and substrate content compared with the expression of pal2. It was further clarified that the environmental stimulation induced down-regulation of both pal2 gene transcription level，increased enzyme activity，which caused the reduction of L-phe content and the increase of t-CA content in the mycelia，and thus negatively regulated the mycelial growth of P. ostreatus.

Key words: Pleurotus ostreatus, stimulation of low-temperature and light, phenylpropanoid metabolic pathway

CLC Number:

S646.141

JIANG Yuqing, HE Qi, HOU Ludan, WU Xiangli, HUANG Chenyang, and ZHAO Mengran. The Molecular Mechanism of Phenylalanine Ammonia-Lyase Gene pal2 Regulating Pleurotus ostreatus Mycelial Growth Under Stimulation of Low-temperature and Light[J]. Acta Horticulturae Sinica, 2023, 50(4): 831-841.

Figures/Tables 6

Table 1 Premer sequences of RT-qPCR

用途Usage	基因Gene	引物序列（5′-3′）Primer sequence	参考文献 Reference
内参基因Reference gene	tublin	F：AGGCTTTCTTGCATTGGTACACGC； R：TATTCGCCTTCTTCCTCATCGGCA	侯志浩等,2019
qPCR	pal1	F：CTCCTTCACAATCGCATCTA；R：CTTCAGCCGCCTATGTTG	Hou et al.,2019
qPCR	pal2	F：CAACTGCTGCGTATGTCA；R：GATGTAGAGGTATGAGGAGATT	Hou et al.,2019

Fig. 1 Effects of environmental stimulation（15 ℃，12 h light/12 h dark）on the growth of Pleurotus ostreatus mycelium

Fig. 2 Effects of environmental stimulation(15 ℃，12 h light/12 h dark)on pal2（A），PAL（B），L-phe（C），and t-CA（D）of Pleurotus ostreatus mycelium Different lowercase letters indicate significant differences among different treatments at 0.05 level. The same below.

Fig. 3 Effect of exogenous addition L-phe and t-CA on the growth of Pleurotus ostreatus mycelium

Fig. 4 Interfering with pal2 facilitates the growth of mycelium under environmental stimulation(15 ℃，12 h light/12 h dark) WT：wild type strain；Ri-1，Ri-9：pal2 interfering strains. The same below.

Fig. 5

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The Molecular Mechanism of Phenylalanine Ammonia-Lyase Gene pal2 Regulating Pleurotus ostreatus Mycelial Growth Under Stimulation of Low-temperature and Light

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