Analysis of the Alternative Splicing Events of Exogenous δ-Aminolevulinic Acid under NaCl Stress in Wild Jujube Seedlings

Abstract

Salt injury, as an abiotic stress factor, seriously affects the quality and yield of crops. This study was conducted by analyzing alternative splicing in the control (CK), δ-aminolevulinic acid (ALA), NaCl, and ALA + NaCl treatments of wild jujube (Zizyphus spinosus (Bunge)Hu) using RNA-seq. It was found that the unique differential alternative splicing is closely related to the alleviation of salt stress and the analyzed intermediates of chlorophyll synthesis and chlorophyll content in the leaves. The results showed that the content and synthesis of chlorophyll were reduced and disrupted in wild jujube leaves under NaCl stress, and the exogenous spraying of ALA could alleviate the effect of NaCl stress on the content of chlorophyll. RNA-seq indicated that the alternative splicing of genes was dominated by exon skipping in all the experimental treatments. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that the CK and ALA + NaCl comparison groups were primarily enriched in porphyrin and chlorophyll metabolism, chloroplast, and energy metabolism pathways. It is hypothesized that ALA treatment can alleviate the effects of salt stress on chlorophyll by altering the alternative splicing of genes related to photosynthesis, chlorophyll metabolism, and energy metabolism in wild jujube. In addition, the verification of alternative splicing genes related to chlorophyll metabolism in wild jujube revealed that treatment with ALA significantly upregulated the expression of HEMH, UROIII, and ChlD genes in the leaves under salt stress and increased the content of the chlorophyll biosynthetic intermediates endogenous ALA, porphobilinogen, protoporphyrin IX, Mg-protoporphyrin IX, and protochlorophyllide, which served to alleviate the effects of NaCl stress on chlorophyll in wild jujube.