Abstract
Alfalfa (Medicago sativa L.) is one of the most important forage crop in northern China, suffers from low-temperature (LT) stress, which significantly impairs its yield. While selenium (Se) supplementation has been reported to enhance abiotic stress tolerance in plants, the underlying molecular mechanisms in alfalfa remain poorly understood. This study aims to explore how Se mediates LT tolerance in alfalfa through transcriptomic analysis. Selenium significantly increased total Se in both shoots and roots, enhanced root-to-shoot translocation, and under low temperature (LT) shifted Se speciation toward organic forms at the higher dose. Meanwhile, 0.4 mg kg−1 Se was sufficient to restore Pn, Gs, Ci, and Tr, reduce MDA, and elevate SOD, POD, APX, GR, GSH, and ASA, indicating improved photosynthetic performance and antioxidant status. Co-expression network analysis identified modules tightly associated with these traits: in the photosynthetic module, MS.gene064837 and MS.gene051360 were positively correlated with Gs, whereas MS.gene061853 and MS.gene037454 were positively correlated with Ci and Tr but negatively with Gs, suggesting coordination between carbon assimilation and stomatal control. In the antioxidant module, MS.gene036174 correlated positively with GSH and MDA and negatively with GSSG, while MS.gene064490 showed negative correlations with Pro, SS, GSSG, SOD, and POD. qRT-PCR of six targets corroborated the RNA-seq trends. Collectively, these results indicate that moderate soil Se (0.4 mg·kg−1) supplementation enhances LT tolerance in alfalfa by concurrently optimizing photosynthetic regulation and antioxidant defense, with MS.gene064837, MS.gene051360, MS.gene036174, and MS.gene064490 emerging as candidate regulators whose specific functions require further validation. This study reveals that selenium enhances LT tolerance in alfalfa by promoting Se conversion into its organic form, optimizing redox homeostasis, and activating key genes associated with photosynthesis and antioxidant defense. These findings provide valuable insights into Se-mediated cold tolerance mechanisms, offering a foundation for Se-based strategies in improving the resilience of forage crops to cold stress.
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Claims 16
Claim · model-written summary
Selenium increased total selenium in alfalfa shoots and roots.
- Taxon
- Alfalfa
Medicago sativae0 - Organ / stage
- shoots and rootse1
- Treatment
- Seleniume1
- Outcome
- total See1
- Direction
- increasede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“Selenium significantly increased total Se in both shoots and roots”
Claim · model-written summary
Selenium enhanced root-to-shoot translocation.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- Seleniume2
- Outcome
- root-to-shoot translocatione1
- Direction
- enhancede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“enhanced root-to-shoot translocation”
e2“Selenium significantly increased total Se in both shoots and roots”
Claim · model-written summary
Higher selenium dose under low temperature shifted selenium speciation toward organic forms.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- Seleniume2
- Outcome
- Se speciatione1
- Direction
- shiftede1
- Qualifier
- under low temperature (LT)e1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“under low temperature (LT) shifted Se speciation toward organic forms at the higher dose”
e2“Selenium significantly increased total Se”
Claim · model-written summary
At low temperature, 0.4 mg kg−1 selenium restored Pn, Gs, Ci, and Tr.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- 0.4 mg kg−1 See1
- Outcome
- Pn, Gs, Ci, and Tre1
- Direction
- restoree1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“0.4 mg kg−1 Se was sufficient to restore Pn, Gs, Ci, and Tr”
Claim · model-written summary
The selenium dose reduced MDA.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- 0.4 mg kg−1 See1
- Outcome
- MDAe1
- Direction
- reducee1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“0.4 mg kg−1 Se was sufficient to restore Pn, Gs, Ci, and Tr, reduce MDA”
Claim · model-written summary
The selenium dose elevated SOD, POD, and APX.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- 0.4 mg kg−1 See1
- Outcome
- SOD, POD, APXe1
- Direction
- elevatee1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“0.4 mg kg−1 Se was sufficient to restore Pn, Gs, Ci, and Tr, reduce MDA, and elevate SOD, POD, APX, GR, GSH, and ASA”
Claim · model-written summary
The selenium dose elevated GR, GSH, and ASA.
- Taxon
- AlfalfaMedicago sativae0
- Treatment
- 0.4 mg kg−1 See1
- Outcome
- GR, GSH, and ASAe1
- Direction
- elevatee1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“0.4 mg kg−1 Se was sufficient to restore Pn, Gs, Ci, and Tr, reduce MDA, and elevate SOD, POD, APX, GR, GSH, and ASA”
Claim · model-written summary
MS.gene064837 and MS.gene051360 were positively correlated with Gs.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene064837 and MS.gene051360e1
- Outcome
- Gse1
- Direction
- positively correlatede1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene064837 and MS.gene051360 were positively correlated with Gs”
Claim · model-written summary
MS.gene061853 and MS.gene037454 correlated positively with Ci and Tr.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene061853 and MS.gene037454e1
- Outcome
- Ci and Tre1
- Direction
- positively correlatede1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene061853 and MS.gene037454 were positively correlated with Ci and Tr but negatively with Gs”
Claim · model-written summary
MS.gene061853 and MS.gene037454 correlated negatively with Gs.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene061853 and MS.gene037454e1
- Outcome
- Gse1
- Direction
- negativelye1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene061853 and MS.gene037454 were positively correlated with Ci and Tr but negatively with Gs”
Claim · model-written summary
MS.gene036174 correlated positively with GSH and MDA.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene036174e1
- Outcome
- GSH and MDAe1
- Direction
- positivelye1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene036174 correlated positively with GSH and MDA and negatively with GSSG”
Claim · model-written summary
MS.gene036174 correlated negatively with GSSG.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene036174e1
- Outcome
- GSSGe1
- Direction
- negativelye1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene036174 correlated positively with GSH and MDA and negatively with GSSG”
Claim · model-written summary
MS.gene064490 correlated negatively with Pro, SS, and GSSG.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene064490e1
- Outcome
- Pro, SS, GSSGe1
- Direction
- negative correlationse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene064490 showed negative correlations with Pro, SS, GSSG, SOD, and POD”
Claim · model-written summary
MS.gene064490 correlated negatively with SOD and POD.
- Taxon
- AlfalfaMedicago sativae0
- Gene
- MS.gene064490e1
- Outcome
- SOD, and PODe1
- Direction
- negative correlationse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“MS.gene064490 showed negative correlations with Pro, SS, GSSG, SOD, and POD”
Claim · model-written summary
qRT-PCR trends for six targets agreed with RNA-seq trends.
- Taxon
- AlfalfaMedicago sativae0
- Outcome
- RNA-seq trendse1
- Comparator
- qRT-PCR of six targetse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Alfalfa (Medicago sativa L.)”
e1“qRT-PCR of six targets corroborated the RNA-seq trends.”
Claim · model-written summary
Moderate soil selenium supplementation enhanced low-temperature tolerance in alfalfa.
- Taxon
- alfalfaMedicago sativae0
- Treatment
- moderate soil Se (0.4 mg·kg−1) supplementatione0
- Outcome
- LT tolerancee0
- Direction
- enhancese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“moderate soil Se (0.4 mg·kg−1) supplementation enhances LT tolerance in alfalfa”
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