Abstract
Dehydrins (DHNs) are late embryogenesis-abundant (LEA) proteins, which enhance abiotic stress tolerance in plants. However, little is known regarding the function of DHNs in Suaeda salsa L. (S. salsa), which can grow in saline soil. Here, we successfully cloned and functionally characterized a dehydrin gene from S. salsa, designated SsDHN. Sequence alignment analysis showed that SsDHN belongs to SKn-type DHNs and shares relatively high level of homology with Spinacia oleracea COR47-like (XP_021846321.1). Quantitative RT-PCR analysis indicated that SsDHN expression level increased significantly under salt stress. We also generated SsDHN-overexpressing transgenic tobacco lines and analyzed their salt stress response. Seeds of transgenic tobacco plants grown under a range of salt concentrations (100, 200, and 300 mM) showed significantly higher germination rates relative to wild-type seeds. Transgenic plants had longer root length, lower relative electrical conductivity (REC), lower malondialdehyde (MDA) content, higher proline (PRO) content, increased peroxidase (SOD) activity, and less damage to the chloroplast ultrastructure. Our results showed that the transgenic tobacco plant lines had improved salt resistance and osmotic adjustment, enhanced reactive oxygen species scavenging ability, maintenance of the K+/Na+ balance, and reduced chloroplast membrane damage. These results suggest that the SsDHN gene may be used for improving abiotic stress tolerance in economically important crops.
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This work was supported by the National Natural Science Foundation of China (Grant No. 31070448) and the Liaoning Province ‘2021 Special Project of Central Government Guiding local scientific technology development’ (2021JH6/10500164). The funding bodies had no roles in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Supplementary file1 (JPG 100 KB)Supplementary Fig. S1 SsDHN clone (847 bp) was PCR amplified from the cDNA of S. salsa leaves via RT-PCR. (A) Products of conserved regions were obtained by RT-PCR. (B) 3′-RACE products were obtained by RT-PCR. (C) The full-length cDNA was obtained by RT-PCR.
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Supplementary file2 (JPG 180 KB)Supplementary Fig. S2 SsDHN sequence information. (A) Nucleotide and deduced amino acid sequences of SsDHN cDNA. Nucleotides are numbered at right. The start (ATG) and stop codons (TAG) in the frames are boxed; (B) Multiple sequence alignment of SsDHN with homologous sequences. Bold lines indicate conserved amino acid sequences of S- and K-segment motifs. SsDHN (AGC55011.1); CqDHN (XP_021756500.1, XP_021732246.1); AcDHN (AFC98463.1); AhDHN (AGZ86543.1); SoDHN (XP_021846321.1); SgDHN (NCBI AEA29617.1); CbDHN (ABV56004.1); AtDHN (CAA62449.1); McDHN (XP_022152554.1); MeDHN (XP_021614140.1); CcDHN (ABC68275.1).
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Supplementary file4 (JPG 86 KB)Supplementary Fig. S3 Molecular analysis of tobacco plant lines. (A) Electrophoregram of PCR products; (B) Southern blot analysis of SsDHN transgene in transgenic tobacco lines; (C) Western blot analysis of SsDHN expression in transgenic tobacco lines. M: DL2000 DNA marker, CK+: the Agrobacterium plasmid transformed into the PBI121-DHN recombinant vector was used as the positive control, CK-: WT tobacco, 1-6: transgenic tobacco.
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Li, H., Zhang, L., Lin, J. et al. SsDHN, a dehydrin protein from Suaeda salsa, enhances salt stress tolerance in transgenic tobacco plants. Plant Growth Regul 99, 299–312 (2023). https://doi.org/10.1007/s10725-022-00908-8
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DOI: https://doi.org/10.1007/s10725-022-00908-8