Abstract
Spinach (Spinacia oleracea) has cold tolerant but heat sensitive characteristics. The spinach variety ‘Island,’ is suitable for summer periods. There is lack molecular information available for spinach in response to heat stress. In this study, high throughput de novo transcriptome sequencing and gene expression analyses were carried out at different spinach variety ‘Island’ leaves (grown at 24 °C (control), exposed to 35 °C for 30 min (S1), and 5 h (S2)). A total of 133,200,898 clean reads were assembled into 59,413 unigenes (average size 1259.55 bp). 33,573 unigenes could match to public databases. The DEG of controls vs S1 was 986, the DEG of control vs S2 was 1741 and the DEG of S1 vs S2 was 1587. Gene Ontology (GO) and pathway enrichment analysis indicated that a great deal of heat-responsive genes and other stress-responsive genes were identified in these DEGs, suggesting that the heat stress may have induced an extensive abiotic stress effect. Comparative transcriptome analysis found 896 unique genes in spinach heat response transcript. The expression patterns of 13 selected genes were verified by RT-qPCR (quantitative real-time PCR). Our study found a series of candidate genes and pathways that may be related to heat resistance in spinach.
Highlights
Agricultural production is threatened by abiotic stresses such as drought, extreme temperatures, flooding and chemical toxicity[1]
The length distribution of Arabidopsis and rice (Oryza sativa) transcriptome sequences were obtained from publicly available datasets
There were 550 and 1131 up-regulated unigenes in the S1 and S2 samples, respectively. In these identified heat-responsive genes, calcium signaling molecule and some transcription factors play an important role in response to early heat stress
Summary
Agricultural production is threatened by abiotic stresses such as drought, extreme temperatures, flooding and chemical toxicity[1]. NGS-based RNA-Seq methods, which are used for transcriptome analysis, even allow simultaneous acquisition of sequences[6] These fast sequencing methods are used to characterize genes, detect gene expression pattern and level, recognize and quantify rare transcripts without prior information of the particular gene or reference genome, and generate information on alternative splicing and sequence variations in identified genes[6,7]. RNA-Seq has been successfully used to reveal the complex biotic and abiotic stress response mechanism in many plant species, such as grape (Vitis vinifera)[8], cotton (Gossypium hirsutum)[9], cucumber (Cucumis sativus)[10], Ammopiptanthus mongolicus[11], and Arabidopsis thaliana[12]. The genome-wide analysis of gene expression during heat treatment was performed on the ‘Island’ summer spinach variety via a NGS-based Illumina paired-end sequencing platform. This study will aid in understanding the heat stress response in spinach at the molecular level, and enriches the genomic information for spinach
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