Abstract
Salt stress is one of the major abiotic factors that affect the metabolism, growth and development of plants, and soybean [Glycine max (L.) Merr.] germination is sensitive to salt stress. Thus, to ensure the successful establishment and productivity of soybeans in saline soil, the genetic mechanisms of salt tolerance at the soybean germination stage need to be explored. In this study, a population of 184 recombinant inbred lines (RILs) was utilized to map quantitative trait loci (QTLs) related to salt tolerance. A major QTL related to salt tolerance at the soybean germination stage named qST-8 was closely linked with the marker Sat_162 and detected on chromosome 8. Interestingly, a genome-wide association study (GWAS) identified several single nucleotide polymorphisms (SNPs) significantly associated with salt tolerance in the same genetic region on chromosome 8. Resequencing, bioinformatics and gene expression analyses were implemented to identify the candidate gene Glyma.08g102000, which belongs to the cation diffusion facilitator (CDF) family and was named GmCDF1. Overexpression and RNA interference of GmCDF1 in soybean hairy roots resulted in increased sensitivity and tolerance to salt stress, respectively. This report provides the first demonstration that GmCDF1 negatively regulates salt tolerance by maintaining K+-Na+ homeostasis in soybean. In addition, GmCDF1 affected the expression of two ion homeostasis-associated genes, salt overly sensitive 1 (GmSOS1) and Na+/H+ exchanger 1 (GmNHX1), in transgenic hairy roots. Moreover, a haplotype analysis detected ten haplotypes of GmCDF1 in 31 soybean genotypes. A candidate-gene association analysis showed that two SNPs in GmCDF1 were significantly associated with salt tolerance and that Hap1 was more sensitive to salt stress than Hap2. The results demonstrated that the expression level of GmCDF1 was negatively correlated with salt tolerance in the 31 soybean accessions (r = -0.56, P < 0.01). Taken together, these results not only indicate that GmCDF1 plays a negative role in soybean salt tolerance but also help elucidate the molecular mechanisms of salt tolerance and accelerate the breeding of salt-tolerant soybean.
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Claims 11
Claim · model-written summary
qST-8 was mapped near Sat_162 on chromosome 8.
- Taxon
- soybean
Glycine maxe0 - Gene
- qST-8e0
- Outcome
- salt tolerancee0
- Qualifier
- chromosome 8e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“A major QTL related to salt tolerance at the soybean germination stage named qST-8 was closely linked with the marker Sat_162 and detected on chromosome 8.”
Claim · model-written summary
GWAS found several SNPs associated with tolerance in the same chromosome 8 region.
- Taxon
- soybean
Glycine maxt0 - Gene
- single nucleotide polymorphisms (SNPs)e0
- Outcome
- salt tolerancee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“a genome-wide association study (GWAS) identified several single nucleotide polymorphisms (SNPs) significantly associated with salt tolerance in the same genetic region on chromosome 8.”
t0“soybean”
Claim · model-written summary
Glyma.08g102000 was identified as a candidate gene and named GmCDF1.
- Taxon
- soybean
Glycine maxt0 - Gene
- Glyma.08g102000e0
- Outcome
- candidate genee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“identify the candidate gene Glyma.08g102000, which belongs to the cation diffusion facilitator (CDF) family and was named GmCDF1.”
t0“soybean”
Claim · model-written summary
GmCDF1 overexpression increased salt sensitivity.
- Taxon
- soybean
Glycine maxe0 - Treatment
- Overexpressione0
- Gene
- GmCDF1e0
- Outcome
- sensitivitye0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“Overexpression and RNA interference of GmCDF1 in soybean hairy roots resulted in increased sensitivity and tolerance to salt stress, respectively.”
Claim · model-written summary
GmCDF1 RNA interference increased salt tolerance.
- Taxon
- soybean
Glycine maxe0 - Treatment
- RNA interferencee0
- Gene
- GmCDF1e0
- Outcome
- tolerance to salt stresse0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“Overexpression and RNA interference of GmCDF1 in soybean hairy roots resulted in increased sensitivity and tolerance to salt stress, respectively.”
Claim · model-written summary
GmCDF1 negatively regulates salt tolerance through K+-Na+ homeostasis.
- Taxon
- soybean
Glycine maxe0 - Gene
- GmCDF1e0
- Outcome
- salt tolerancee0
- Qualifier
- maintaining K+-Na+ homeostasise0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“GmCDF1 negatively regulates salt tolerance by maintaining K+-Na+ homeostasis in soybean.”
Claim · model-written summary
GmCDF1 affected expression of GmSOS1 and GmNHX1 in transgenic roots.
- Taxon
- soybean
Glycine maxt0 - Gene
- GmSOS1e0
- Outcome
- expression of two ion homeostasis-associated genese0
- Qualifier
- transgenic hairy rootse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“GmCDF1 affected the expression of two ion homeostasis-associated genes, salt overly sensitive 1 (GmSOS1) and Na+/H+ exchanger 1 (GmNHX1), in transgenic hairy roots.”
t0“soybean”
Claim · model-written summary
Ten GmCDF1 haplotypes were detected in 31 genotypes.
- Taxon
- soybean
Glycine maxe0 - Gene
- GmCDF1e0
- Outcome
- haplotypes of GmCDF1e0
- Value
- ten haplotypese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“a haplotype analysis detected ten haplotypes of GmCDF1 in 31 soybean genotypes.”
Claim · model-written summary
Two GmCDF1 SNPs were significantly associated with tolerance.
- Taxon
- soybean
Glycine maxt0 - Gene
- GmCDF1e0
- Outcome
- salt tolerancee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“two SNPs in GmCDF1 were significantly associated with salt tolerance”
t0“soybean”
Claim · model-written summary
Hap1 was more salt-sensitive than Hap2.
- Taxon
- soybean
Glycine maxt0 - Material
- Hap1e0
- Outcome
- sensitive to salt stresse0
- Direction
- more sensitivee0
- Comparator
- Hap2e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“Hap1 was more sensitive to salt stress than Hap2.”
t0“soybean”
Claim · model-written summary
GmCDF1 expression correlated negatively with tolerance in 31 accessions.
- Taxon
- soybean
Glycine maxe0 - Gene
- GmCDF1e0
- Outcome
- salt tolerancee0
- Direction
- negatively correlatede0
- Value
- r = -0.56e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.

Verbatim
e0“the expression level of GmCDF1 was negatively correlated with salt tolerance in the 31 soybean accessions (r = -0.56, P < 0.01).”
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