Genome-wide analysis of the annexin gene family in alfalfa (Medicago sativa L.) and functional characterization of MsANN22 under drought stress
- 1Shihezi University
Abstract
Annexins are a conserved family of calcium-dependent phospholipid-binding proteins that participate in diverse plant stress responses. However, the annexin gene family in Medicago sativa and the molecular mechanisms underlying annexin-mediated drought tolerance remain largely unexplored. In this study, 73 MsANN genes were identified in the alfalfa genome. Their phylogenetic relationships, synteny, conserved motifs, gene structures, and promoter cis-acting elements were systematically analyzed. Synteny analysis revealed 75 segmental/whole-genome duplication-associated gene pairs but no tandem duplications, indicating that large-scale duplication was the major driver of MsANN family expansion. The MsANN promoters contained ABA- and drought-responsive elements. Expression profiling identified MsANN22 as a strongly drought-responsive gene encoding a cytoplasm-localized protein. MsANN22 overexpression enhanced drought tolerance in both Arabidopsis thaliana and alfalfa: in A. thaliana, MsANN22 overexpression promoted seed germination under osmotic stress, whereas in alfalfa it enhanced antioxidant capacity, with superoxide dismutase and peroxidase activities reaching 1.38- and 1.70-fold those of wild-type plants, respectively. Consistently, MsANN22-overexpressing alfalfa accumulated approximately 20% less malondialdehyde than wild-type plants and exhibited reduced H2O2 and O2*- accumulation, as indicated by DAB and NBT staining. Yeast two-hybrid and luciferase complementation assays further supported physical interactions of MsANN22 with MsIAA and MsPR-10. These findings provide insight into the molecular mechanism by which MsANN22 mediates drought tolerance and offer a candidate gene for improving drought tolerance in alfalfa.
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BibTeX
@misc{xie2026genomewide,
title = {{Genome-wide analysis of the annexin gene family in alfalfa (Medicago sativa L.) and functional characterization of MsANN22 under drought stress}},
author = {Q. Xie and j. guangzhi and J. Wang and h. he and p. du and h. li and Z. Meng and q. shao and h. li},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.21.760862},
url = {https://www.biorxiv.org/content/10.64898/2026.10.21.760862v1},
note = {CC BY-ND},
}
RIS
TY - UNPB
TI - Genome-wide analysis of the annexin gene family in alfalfa (Medicago sativa L.) and functional characterization of MsANN22 under drought stress
AU - Q. Xie
AU - j. guangzhi
AU - J. Wang
AU - h. he
AU - p. du
AU - h. li
AU - Z. Meng
AU - q. shao
AU - h. li
PY - 2026
DA - 2026/10/21
PB - bioRxiv
DO - 10.64898/2026.10.21.760862
UR - https://www.biorxiv.org/content/10.64898/2026.10.21.760862v1
AB - Annexins are a conserved family of calcium-dependent phospholipid-binding proteins that participate in diverse plant stress responses. However, the annexin gene family in Medicago sativa and the molecular mechanisms underlying annexin-mediated drought tolerance remain largely unexplored. In this study, 73 MsANN genes were identified in the alfalfa genome. Their phylogenetic relationships, synteny, conserved motifs, gene structures, and promoter cis-acting elements were systematically analyzed. Synteny analysis revealed 75 segmental/whole-genome duplication-associated gene pairs but no tandem duplications, indicating that large-scale duplication was the major driver of MsANN family expansion. The MsANN promoters contained ABA- and drought-responsive elements. Expression profiling identified MsANN22 as a strongly drought-responsive gene encoding a cytoplasm-localized protein. MsANN22 overexpression enhanced drought tolerance in both Arabidopsis thaliana and alfalfa: in A. thaliana, MsANN22 overexpression promoted seed germination under osmotic stress, whereas in alfalfa it enhanced antioxidant capacity, with superoxide dismutase and peroxidase activities reaching 1.38- and 1.70-fold those of wild-type plants, respectively. Consistently, MsANN22-overexpressing alfalfa accumulated approximately 20% less malondialdehyde than wild-type plants and exhibited reduced H2O2 and O2*- accumulation, as indicated by DAB and NBT staining. Yeast two-hybrid and luciferase complementation assays further supported physical interactions of MsANN22 with MsIAA and MsPR-10. These findings provide insight into the molecular mechanism by which MsANN22 mediates drought tolerance and offer a candidate gene for improving drought tolerance in alfalfa.
ER -