Evaluation of drought resistance and transcriptome analysis for the identification of drought-responsive genes in Iris germanica

Iris germanica, a species with very high ornamental value, exhibits the strongest drought resistance among the species in the genus Iris, but the molecular mechanism underlying its drought resistance has not been evaluated. To investigate the gene expression profile changes exhibited by high-drought-resistant I. germanica under drought stress, 10 cultivars with excellent characteristics were included in pot experiments under drought stress conditions, and the changes in the chlorophyll (Chl) content, plasma membrane relative permeability (RP), and superoxide dismutase (SOD), malondialdehyde (MDA), free proline (Pro), and soluble protein (SP) levels in leaves were compared among these cultivars. Based on their drought-resistance performance, the 10 cultivars were ordered as follows: ‘Little Dream’ > ‘Music Box’ > ‘X’Brassie’ > ‘Blood Stone’ > ‘Cherry Garden’ > ‘Memory of Harvest’ > ‘Immortality’ > ‘White and Gold’ > ‘Tantara’ > ‘Clarence’. Using the high-drought-resistant cultivar ‘Little Dream’ as the experimental material, cDNA libraries from leaves and rhizomes treated for 0, 6, 12, 24, and 48 h with 20% polyethylene glycol (PEG)-6000 to simulate a drought environment were sequenced using the Illumina sequencing platform. We obtained 1, 976, 033 transcripts and 743, 982 unigenes (mean length of 716 bp) through a hierarchical clustering analysis of the resulting transcriptome data. The unigenes were compared against the Nr, Nt, Pfam, KOG/COG, Swiss-Prot, KEGG, and gene ontology (GO) databases for functional annotation, and the gene expression levels in leaves and rhizomes were compared between the 20% PEG-6000 stress treated (6, 12, 24, and 48 h) and control (0 h) groups using DESeq2. 7849 and 24,127 differentially expressed genes (DEGs) were obtained from leaves and rhizomes, respectively.

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Fig: Gene ontology classification of assembled unigenes (a). KOG function classification of assembled unigenes (b). Functional classification and pathway assembled unigenes by KEGG (c); The A–E indicates Cellular Processes, Environmental Information Processing, Genetic Information Processing, Metabolism, Organismal Systems, respectively.

GO and KEGG enrichment analyses of the DEGs revealed significantly enriched KEGG pathways, including ribosome, photosynthesis, hormone signal transduction, starch and sucrose metabolism, synthesis of secondary metabolites, and related genes, such as heat shock proteins (HSPs), transcription factors (TFs), and active oxygen scavengers. In conclusion, we conducted the first transcriptome sequencing analysis of the I. germanica cultivar ‘Little Dream’ under drought stress and generated a large amount of genetic information. This study lays the foundation for further exploration of the molecular mechanisms underlying the responses of I. germanica to drought stress and provides valuable genetic resources for the breeding of drought-resistant plants.

Zhang, J., Huang, D., Zhao, X. et al. Evaluation of drought resistance and transcriptome analysis for the identification of drought-responsive genes in Iris germanicaSci Rep 11, 16308 (2021). https://doi.org/10.1038/s41598-021-95633-z

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