Supplementary Table 1. Frequency of MSH1-dr sorghum plants arising in T3 families. Eleven individual T3 families, all containing the MSH1-RNAi transgene, were evaluated for presence of the MSH1-dr phenotype (dwarfed stature, enhanced tillering, delayed flowering). In these cases, the identified MSH1-dr plants did not flower without GA application. All T3 families are from a single transformation event.

FamilyNo.PlantsNo. MSH1-dr*

T3-1290

T3-2102 (20%)

T3-3173 (18%)

T3-4 80

T3-5 40

T3-6140

T3-7349 (26%)

T3-8 60

T3-9180

T3-10250

T3-11183 (17%)

Total18317 (9.3%)

Supplementary Table 2. Mean stomatal number per sample in sorghum non-transgenic MSH1-dr
plants versus wildtype TX430. Counts were taken from adaxial and abaxial surfaces of each leaf.
Stomatal numbers were significantly lower (P<0.0001) between TX430 and the MSH1-dr plants
for both adaxial and abaxial readings.
Adaxial / Abaxial
Genotype / N / Mean / StdDev / N / Mean / StdDev
TX430 / 20 / 41 / 6 / 20 / 54 / 11
MSH1-dr / 18 / 21 / 3 / 18 / 30 / 4

Supplementary Table 3.Plant height measurements of MSH1-dr x Tx430 direct (A) and reciprocal crosses (B). A and B experiments were conducted separately. All MSH1-dr plants, displaying the altered growth phenotype, were confirmed to lack the transgene.

  1. Plant height of MSH1-dr x Tx430 F1 and parental populations

Genotype N / Mean
(cm) / Std Error
GAII11 x TX430 / 7 / 172.4*** / 2.1
GAII15 x TX430 / 15 / 136.8*** / 7.5
GAII22 x TX430 / 5 / 122.2 / 17.3
GAII24 x TX430 / 19 / 113.3* / 6.4
GAII25 x TX430 / 12 / 121.4*** / 6.7
GAII28 x TX430 / 13 / 124.2** / 9.8
GAII11 / 17 / 59.6*** / 4.5
GAII15 / 19 / 45.1*** / 5.6
GAII22 / 18 / 56.1*** / 4.4
GAII24 / 17 / 45.8*** / 4.7
GAII25 / 15 / 44.0*** / 4.6
GAII28 / 18 / 56.5*** / 4.1
TX430 / 16 / 97.6 / 2.3

Differences relative to TX430 are significant at * p<0.05, **p<0.01, and *** p<.001

  1. Plant height in reciprocal crosses of sorghum TX430 x MSH1-dr populations

Genotype / N / Mean (cm) / Std Error
TX430 x GAII22 / 8 / 142.8*** / 10.4
TX430 x GAII23 / 8 / 123.1 / 9.7
TX430 x GAII44 / 6 / 172.2*** / 3.9
GAII22 / 4 / 50.8*** / 12.5
GAII23 / 5 / 40.0*** / 6.3
GAII44 / 4 / 34.0*** / 1.8
TX430 / 13 / 107.5 / 3.7

*** Differences relative to TX430 are significant at p<0.0001

Supplementary Table 4. Primers used in quantitative PCR and hybridization assays

MIR156a-F5'- CTC AAG TTC ATT GCC ATT TTT AGG -3'

MIR156a-R5'- GAG AGA TTG AGA CAT AGA GAA CGA AGA -3'

FLC-F5'- GGA TGC GTCA CAG AGA ACA G-3'

FLC-R5'- CGA CAA GTC ACC TTC TCC AA-3'

SVP-F5'- TGA ATC GGA GAA CGC TGC TGT GTA -3'

VP-R5'- TCT AAC CAC CAT ACG GTA AGC CGA -3'

SOC1-F5'- TAA GGA TCG AGT CAG CAC CAA ACC -3'

SOC1-R5'- AGC ATG TTC CTA TGC CTT CTC CCA -3'

GA2OX6-F5'- TGAATCACTATCCACCAGCACCGT

GA2OX6-R5'- AAGGCAGTCACCGACCAATACGAA

CYCP4;1-F5'- ATC ACT AGT GTC ATG GTC GCT GCT -3'

CYCP4;1-R5'- GTA GGC GTT GAA TGT GTT TGG CGT -3'

EXPA8-F5'- TCA ACC ATC ACC GTC ACA GCT ACA -3'

EXPA8-R5'- ATG CTG AAG AGG AGG ATT GCA CCA -3'

GASA6-F 5'- TGT GGA GGA CAA TGC ACA AGG AGA -3'

GASA6-R5'- GGA GGG ACA CAA AGG CAT TTA GCA -3'

PIN1-F5'- TGC TCG TTG CTT CTT ATG CCG TTG -3'

PIN1-R5'- ACC GCA GTG CTA AGA ATG TCA GGA -3'

PIN7-F5'- TTG GGC TCT TGT TGC TTT CAG GTG -3'

PIN7-R5'- TGC CAT ACC AAG ACC AGC ATC AGA -3'

IAA7-F5'- ACG TTT CTG CTG TTC CCA AGG AGA -3'

IAA7-R5'- ACC ACT ACT GGT CTT CTG CTG AGT -3'

CYP79B3-F5'- ACG ACC GTC GCA GGT TAC CAT ATT -3'

CYP79B3-R5'- AGC ACA TCC TCT CTT TCC GGT ACT -3'

SorMshIRt-Fu1 5’- TGG CTA CTC AAT AGG AGG CAG GAA -3’

SorMshIRt-Ru1 5’- TAT GCA CAA GGC TAG CAC CAC TGA -3’

Sb10g028020UBQ10-F 5'- TTG TGA AGA CCC TCA CTG GCA AGA -3’

Sb10g028020UBQ10-R 5'- AAT CAG CAA GGG TAC GAC CAT CCT -3’

3-2L5’- AAACAACCAAAAAAAAAAACATACCT-3’

3-2R5’-TGTTATTTTTATTGTAAAGATTTAATAGTTT-3’

3-2Ln 5’-CAAAAAAACTAAAAAACTATATTTAACT-3’

Probes for small RNA gel blot analysis

U65’TCATCCTTGCGCAGGGGCCA3’

miR1565’gtGctCacTctCttCtgTca3’

miR1725’ATGCAGCATCATCAAGATTCT3’

Assay for sorghum RNAi transgene

Intron-rev 5’GTGTACTCATCTGGATCTGTATTG3’

msf85’GGTTGAGGAGCCTGAATCTCTGAAGA3’