This research centers on the sand-fixing plant known as Stipagrostis pennata, from which the β-glucosidase gene SpBGLU25 was successfully cloned using the molecular cloning method. SpBGLU25 encodes a hydrophilic and stable protein made up of 193 amino acids, located in the cell membrane. qRT-PCR analysis indicated that the expression of the SpBGLU25 is closely linked to the drought stress tolerance of S. pennata. Following this, functional validation was performed using an Arabidopsis overexpression system. The overexpression of transgenic Arabidopsis lines showed significantly improved drought tolerance under PEG and mannitol treatments. Assessments of germination, root length, and physiological indicators such as proline, malondialdehyde content, soluble sugars, and relative leaf water content (RLWC) further confirmed the enhanced performance of the overexpressing plants. Additionally, the comparative transcriptomic analysis of SpBGLU25-OE Arabidopsis compared to the wild-type (WT) showed that differentially upregulated genes were primarily enriched in categories of “cellular process,” “cell,” and “catalytic activity.” KEGG pathway enrichment analysis indicated that the genes were mainly concentrated in the pathways of phenylpropanoid biosynthesis and plant hormone signal transduction. These findings provide a crucial foundation for further investigation into the function of the SpBGLU25 and its role in regulating plant tissue development and adaptation to stress. This research is anticipated to offer new theoretical insights and genetic resources for enhancing plant stress tolerance through genetic engineering.

Lipid transfer proteins (LTPs) play crucial regulatory roles in plant growth, development, and abiotic stress responses. Stipagrostis pennata is a species of grass widely distributed in arid and semi-arid regions, particularly adapted to desert and steppe environments. Under extreme drought conditions, it exhibits a variety of physiological and morphological adaptation mechanisms, making it an important species for studying plant drought tolerance. Recently, LTPs have been found to exhibit upregulated expression under drought stress in plants such as wheat and tobacco, enhancing their drought tolerance. However, the functional role of LTPs in S. pennata remains unexplored. In this study, the SpLTP1 gene was isolated from S. pennata via molecular cloning, encoding a 116-amino acid protein. Phylogenetic analysis revealed that this protein contains a highly conserved nsLTP1 (cd01960) domain and has high sequence similarity with LTPs of Setaria viridis, Setaria italica, Musa acuminata and Phragmites australis. qRT-PCR revealed that SpLTP1 was highly expressed and dynamically regulated under drought, suggesting its potential role in root rhizosheath formation and drought tolerance. To investigate SpLTP1 function, SpLTP1-overexpressing (SpLTP1-OE) and complementation (SpLTP1-atltp) Arabidopsis lines were generated using the floral dip method, in comparison with the existing wild-type (WT) and the LTP-deficient mutant (atltp). Drought stress phenotyping and physiological assays indicated that SpLTP1 likely enhances drought tolerance by elevating antioxidant enzyme activities and osmolyte accumulation. Comparative transcriptome analysis of SpLTP1-OE and WT plants further suggested that SpLTP1 modulates critical pathways, including phenylpropanoid biosynthesis, zeatin biosynthesis, and plant hormone signal transduction, thereby influencing plant growth and stress adaptation. These findings not only provide novel insights into the molecular mechanisms by which SpLTP1 regulates rhizosheath development in S. pennata but also establish a foundation for deciphering its role in extreme drought adaptation.

Sugars Will Eventually Be Exported Transporters (SWEETs) are involved in plant growth and development, particularly in resistance to adverse environments. Achnatherum splendens (Trin.) Nevski exhibits rhizosheath formation and demonstrates notable salt and drought tolerance. We identified 31 sugar transporter family genes (AsSWEETs) from the Achnatherum splendens genome in the NCBI database and performed bioinformatics analyses, including gene structure, subcellular localization, conserved sequences, promoter cis-acting elements, phylogenetic relationships, and chromosomal localization. The 31 AsSWEET genes are distributed across 13 chromosomes, encoding peptides ranging from 375 to 1353 amino acids. Their predicted molecular weights range from 31,499.38 to 109,286.91 Da, with isoelectric points (pI) between 4.78 and 5.21. The aliphatic index values range from 13.59 to 24.19, and the grand average of hydropathicity (GRAVY) values range from 0.663 to 1.664. An analysis of promoter cis-acting elements reveals that all 31 AsSWEET genes contain multiple elements related to light, stress, and hormone responses. Subcellular localization predictions indicate that most genes in this family are localized to the plasma membrane or tonoplast, with AsSWEET12-2 and AsSWEET3b localized in chloroplasts and AsSWEET2b-2 in the nucleus. qRT-PCR results show that AsSWEET13-1, AsSWEET13-3, and AsSWEET1a exhibit upregulated expression in response to salt and drought stress in the roots of Achnatherum splendens. These genes may serve as candidate genes for investigating the stress resistance mechanisms of Achnatherum splendens. The findings provide a theoretical basis for further research on stress resistance mechanisms and candidate gene identification under salt and drought stress in Achnatherum splendens.