Lushan Botanical Garden, Jiangxi Province and Chinese Academy of Sciences, Jiujiang, 332900, Jiangxi, China.
College of Life Science, Nanchang University, Nanchang, 330031, Jiangxi, China.
Sci Rep. 2024 Jun 25;14(1):14588. doi: 10.1038/s41598-024-65555-7.
Autophagy is a highly conserved eukaryotic pathway and plays a crucial role in cell survival under stress conditions. Here, we applied a full-length transcriptome approach to study an Arabidopsis autophagy mutant (atg5-1) subjected to nitrogen-starvation, using Oxford Nanopore Technologies. A total of 39,033 transcripts were identified, including 11,356 new transcripts. In addition, alternative splicing (AS) events and lncRNAs were also detected between Col-0 (WT) and atg5-1. Differentially expressed transcript enrichment showed that autophagy upregulates the expression of many stress-responsive genes and inhibits the transcription of photosynthesis-associated genes. The qRT-PCR results showed that the expression patterns of photosynthesis-related genes in the atg5-1 differed under the conditions of nitrogen starvation and carbon starvation. Under nitrogen starvation treatment, many genes related to photosynthesis also exhibited AS. Chlorophyll fluorescence images revealed that the Fv/Fm and ΦPSII of old atg5-1 leaves were significantly reduced after nitrogen starvation treatment, but the Y(NPQ) indices were significantly increased compared to those of the WT plants. The results of qRT-PCR suggest that autophagy appears to be involved in the degradation of genes related to photodamage repair in PSII. Taken together, the full-length transcriptiome sequencing provide new insights into how new transcripts, lncRNAs and alternative splicing (AS) are involved in plant autophagy through full-length transcriptome sequencing and suggest a new potential link between autophagy and photosynthesis.
自噬是一种高度保守的真核途径,在应激条件下的细胞存活中起着至关重要的作用。在这里,我们应用全长转录组方法研究氮饥饿条件下的拟南芥自噬突变体(atg5-1),使用牛津纳米孔技术。总共鉴定出 39033 个转录本,包括 11356 个新转录本。此外,还在 Col-0(WT)和 atg5-1 之间检测到了可变剪接(AS)事件和 lncRNAs。差异表达转录本富集表明,自噬上调了许多应激响应基因的表达,并抑制了光合作用相关基因的转录。qRT-PCR 结果表明,在氮饥饿和碳饥饿条件下,atg5-1 中光合作用相关基因的表达模式不同。在氮饥饿处理下,许多与光合作用相关的基因也表现出 AS。叶绿素荧光图像显示,氮饥饿处理后,老的 atg5-1 叶片的 Fv/Fm 和 ΦPSII 显著降低,但与 WT 植物相比,Y(NPQ)指数显著增加。qRT-PCR 的结果表明,自噬似乎参与了 PSII 中与光损伤修复相关基因的降解。总之,全长转录组测序通过全长转录组测序提供了新的见解,了解新转录本、lncRNAs 和可变剪接(AS)如何参与植物自噬,并提出了自噬和光合作用之间的新的潜在联系。