Abstract：Pleurotus pulmonarius has the characteristics of changing temperature and fruiting. Low temperature stimulation can induce the formation of fruiting body. Using Illumina sequencing technology, the complete transcriptome including four different stages of growth and development: Mycelial culture at normal temperature stage, low temperature induction stage, recovery to normal temperature stage after low temperature induction and primordium formation stage of Pleurotus pulmonarius were sequenced. There were 41 544 496, 45 722 382, 46 642 306 and 49 980 456 clean reads obtained at four different growth stages respectively. The clean reads of four different stages samples transcriptome sequencing were aligned to the reference sequences which were spliced by Trinity software. The alignmengt rate of the four sequencing samples was 83.22%, 82.85%, 83.10% and 82.59%, respectively. Analysis of differentially expressed genes showed that there were 1 216 genes up-regulated while 1 046 of them were down-regulated expressed between two libraries with normal and low-temperature induction. Meanwhile, 1 331 of genes were up-regulated while 1 784 of them were down-regulated in the primordium library compared with library of mycelium induced by low-temperature. Functional annotation with Blast-Nr database indicated that these up-regulated genes in mycelium induced by low-temperature were involved in transferring alkyl, aspartic-type endopeptidase activity, oxidation-reduction process, cell wall integrity and intracellular protein transport. And in primordium, these up-regulated genes were involved in regulation of transcription, ribosome synthetize, methelation and ion channel. Gene Ontology functional enrichment analysis showed that the differentially expressed genes involved in catalytic activity, single organism metabolic process and oxidoreductase activity were significantly enriched at two stages of normal temperature treatment and low temperature induction, while at the stages of recovery to normal temperature stage after low temperature induction and primordium formation catalytic activity, single tissue process and single tissue metabolism process were significantly enriched. KEGG metabolic pathway analysis showed that there were significant differences in gene enrichment in two metabolic pathways: TCA cycle and endoplasmic reticulum protein processing. Through the comparative study of transcriptome in different stages of growth and development, 2 262 and 3 115 differentially expressed genes were screened out. qRT-PCR analysis showed that the expression changes of the selected genes at different stages were consistent with the results of transcriptome sequencing. This study lays the foundation for further excavation of the key genes of primordium formation and fruiting body growth of Pleurotus pulmonarius, and also provides guidance for the application of genetic engineering to cultivate new high-quality and high-yield Pleurotus pulmonarius varieties.
王伟科, 陆娜, 闫静, 宋吉玲, 袁卫东, 周祖法. 秀珍菇不同生长阶段在低温刺激后的转录组及差异基因表达分析[J]. 农业生物技术学报, 2020, 28(10): 1837-1848.
WANG Wei-Ke, LU Na, YAN Jing, SONG Ji-Ling, YUAN Wei-Dong, ZHOU Zu-Fa. Analyses of Transcriptome and Differentially Expressed Genes at Different Developmental Stages of Pleurotus pulmonarius Treated with Low Temperature. 农业生物技术学报, 2020, 28(10): 1837-1848.
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