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Supplemental Table 1
the primers used in this study
Primer name
Sequence
ZmHKT1;1a-F
GTCGT CGTGCTGTATGTGCT
ZmHKT1;1a-R
TACGCACTGATGACCTCGAC
ZmHKT1;1b-F
CGACAATAGCACGAGCAAGA
ZmHKT1;1b-R
TCAGCACGTTGGATTATTGC
ZmGAPDH F
CCCTTCATCACCACGGACTAC
ZmGAPDH R
AACCTTCTTGGCACCACCCT
AY562132-F
GTAGCATTGTTGGTGGTGGTGTG
AY562132-R
ACCGTGGAGCAGTCAATGGAAG
AY554169-F
ACAGTCGTGAGCATTCCCAAC
AY554169-R
CCAAACCTTCTGTGCTACCTC
AY554170-F
CCGGAGTGAAGGGGATGG
AY554170-R
CAAGCAATGTGAATGGTATGTGAG
NtERD10B-F
CAATTTAGTGCAGGCCAGGC
NtERD10B-R
GGTCCATGGTGGCCAGGAAG
NtERD10C-F
GGGTAGCGCAAACGTGGAG
NtERD10C-R
CTTTTCCCTCAGCCTCGTGC
NtSOS1-F
CAAATGTTATCCCCCGAAAGC
NtSOS1-R
CGGAGAACCTGAGGAAATGTGA
NtNHX2-F
ACTCATCCCCATTGGTCCG
NtNHX2-R
AAGGAGTTCCACAAAAGCACGA
NtNHX4-F
CAAGAACTTCCGCACCCAC
NtNHX4-R
GCAGTATCAAACGCAGAGGACC
NtCAX3-F
CGGTTTGGCAATAATTGTCACAG
NtCAX3-R
CAACGATCATGCTTCAATCATCC
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Supplemental Table 2 The stress-related genes
Gene name
GenBank accession number and major function
NtGPX
AB041518；encodes glutathione peroxidase；related to oxidative stress
NtSPS
AF194022；encodes Sucrose-6-phosphate synthase A；related to osmotic
stress
NtERD10B
AB049336；encodes dehydrin；related to osmotic stress
NtRub-SS
X02353；encodes ribulose 1,5-bisphosphate carboxylase small
Subunit；involved in photosynthetic carbon
assimilation/metabolism
NtAPX2
D85912；encodes cytosolic ascorbate peroxidase； related to oxidative
stress
NtCAX3
-;
NtRCA342
NtNHX4
NtAPX1
ChlGaPA
AY554170
vacuolar cation/proton exchanger 3；related to Ca2+ transport
U35111；encodes rubisco activase precursor；involved in photosynthetic
carbon assimilation/metabolism
-；encodes sodium hydrogen exchanger 4；related to Na+ transport
AU15933；encodes cytosolic ascorbate peroxidase；related to oxidative
stress
M14417；encodes A-subunit of chloroplast glyceradehyde-3-P
dehydrogenase；involved in photosynthetic carbon
assimilation/metabolism
AY554170；encodes raffinose synthase family protein/seed imbibitions
protein；related to osmotic stress
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7
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ZmHKT1;1
Actin
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8
9
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12
13
14
15
16
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Supplemental Fig. 1 Vectors construction and molecular identification
(A)Schematic representation of the pCAMBIA3301-35S:ZmHKT1;1 constructs used
to transform tobacco plants with the two transcripts of ZmHKT1;1 gene(ZmHKT
1;1a/b stands for the two transcripts);
(B)Verification of transgenic lines via PCR. Lane 1, H2O as blank control; Lane 2,
WT plants as negative control; Lane 3, transgenic line 1-7 overexpressing ZmHKT
1;1a; Lane 4, transgenic line 1-13 overexpressing ZmHKT 1;1a; Lane 5, transgenic
line 2-3 overexpressing ZmHKT 1;1b; Lane 6, transgenic line 2-8 overexpressing
ZmHKT 1;1b;
(C) RT-PCR analysis of the expression level of ZmHKT 1;1 gene in T2 plants. Lane 1,
H2O as blank control; Lane 2, WT plants as negative control; Lane 3, transgenic line
1-7 overexpressing ZmHKT 1;1a; Lane 4, transgenic line 1-13 overexpressing
ZmHKT 1;1a; Lane 5, transgenic line 2-3 overexpressing ZmHKT 1;1b; Lane 6,
transgenic line 2-8 overexpressing ZmHKT 1;1b;
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Supplemental Fig. 2 Leaf disc assay of WT and transgenic lines (1-7,1-13,2-3 and
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2-8) at different NaCl concentrations (0mM,200mM and 300mM) in T2 plants.
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Supplemental Fig. 3 Na+ and K+ contents in shoot(A and B) and root(C and D) of
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transgenic lines (1-7,1-13,2-3 and 2-8).
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24-day-old WT and T2 transgenic plants were transferred into plastic containers
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containing improved Hoagland nutrient solution and pre-culture 3 days prior to
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receiving salt treatment of 0, 200 and 300mM NaCl. Shoot and root tissues were
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collected after 6 d of treatment and 3 d of recovery. Data are shown as the average ±
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S.E. of 15 plants. Experimental data was analyzed by t-test and the asterisks in
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columns mean significant difference from WT plants at *P<0.05 or **P<0.01 level.
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25
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ZmHKT1;1a
ZmHKT1;1b
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Supplemental Fig. 4 The relative expression changes of ZmHKT1;1a and
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ZmHKT1;1b in different tissues of maize.
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The relative transcriptional level was analyzed using 2-ΔΔCt method, and the
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transcriptional level of ZmHKT1;1a in leaf was normalized as 1.00.
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