Supplementary Material for Biotechnology Letters
Hong et al.
Supplementary material for Biotechnology Letters
Overexpression of Brassica rapa SHI-RELATED SEQUENCE genes suppresses growth and development in Arabidopsis thaliana
Joon Ki Hong · Jin A Kim · Jung Sun Kim · Soo In Lee · Yeon-Hee Lee ( )
National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea
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Supplementary Table 1 Primers of GA-related genes used in RT-PCR
Target genes / AGI a / Primer sequences / Expected size (bp)AGL15 / At5g13790 / 5’-TCGAGAATGCGAATAGCAGACAAG-3’
5’-TTTGTTGTCACTGAATCGCTTGAC-3’ / 681
DDF1 / At1g12610 / 5’-AGAGCCTAAGTCAGAAGAAAGACTC-3’
5’-ATTCCATACCATCAAGGAAACTTC-3’ / 904
RGA2 / At1g14920 / 5’-AGACGACGGTAACGGCATGGATG-3’
5’-AGCTTACACCCAACTTCATGAAG-3’ / 912
PLK / At2g25170 / 5’-TTCACTGCTCTTGGGTGCCTGAG-3’
5’-TTCTGAAGTGGTGTTCCTGTCAG-3’ / 946
SLY1 / At4g24210 / 5’-TGATTGATCTTCGCAATCGATTCC-3’
5’-ACAATCATTGTCCAACTACGGACC-3’ / 762
SNE / At5g48170 / 5’-TCTTGGTTCACCATGTCGTCGGAG-3’
5’-TTCCGGTTACGGTTCCGGTTCCGG-3’ / 529
GAMYB33 / At5g06100 / 5’-TGGAAGTGACTGCAGAAGTAGATG-3’
5’-AACATATCGAGCAGACCAGTATC-3’ / 1062
GAMYB65 / At3g11440 / 5’-ATGAGTTACACGACGGCGACTGC-3’
5’-ATACGTAACTGTATGGCTATCAGG-3’ / 933
a Arabidopsis genome initiative number.
Supplementary Table 2 Primers of auxin-related genes used in RT-PCR
Target genes / AGI a / Primer sequences / Expected size (bp)YUCCA / At4g32540 / 5’-ACTGACCAGACCCAGCATATCATC-3’
5’-ACATAGCTCCGACATCGAGAACCG-3’ / 844
IAMT1 / At5g55250 / 5’-TATGTAACATGAAGCTCGAGAGAC-3’
5’-TTGCAAGCTTGGTGCGTACACCG-3’ / 842
SUR1 / At2g20610 / 5’-TACCTGTATCGAAGCTGAAGACGC-3’
5’-TGAGCTTCGACTCCGATGGTTATC-3’ / 1020
PIN1 / At1g73590 / 5’-TGATCCTCGCTTACGGCTCTGTC-3’
5’-TCCCAGCTCCAGCAGCAGTTCCAG-3’ / 873
IAA3 / At1g04240 / 5’-ACCTCAAGGAAACAGAGCTGAGGC-3’
5’-TTCCAGTTTCATATGCTTAAGACC-3’ / 771
IAA6 / At1g52830 / 5’-AAGTATCAAAACTTCATATAGCC-3’
5’-AGATGATCAACACTAGCTAGTATG-3’ / 652
ARF7 / At5g20730 / 5’-ATCTTCGATTGTCCTTGGTCATCC-3’
5’-TGAAGAGTCATCTGCGCATAGACC-3 / 822
AXR1 / At1g05180 / 5’-ACCAAGTACGATCGTCAGCTCAGG-3’
5’-AGGTCATATCTGGTATAGAACCTTC-3’ / 952
Actin / At3g18780 / 5′-ATGGCTGAGGCTGATGATATTCAA-3′
5′-TTAGAAACATTTTCTGTGAACGATT-3′ / 1220
a Arabidopsis genome initiative number.
Supplementary Fig. 1 The expression of BrSRS genes in transgenic Arabidopsis. (a) Structure of the 35S-Pro::BrSRS constructs for expression of BrSRS genes. The gene encoding bacterial neomycin phosphotransferase II (npt II), which is regulated by the nopaline synthase gene promoter (nos-pro) and 3’ terminator (nos-ter), serves as a selectable marker for Arabidopsis transformation. BrSRS genes are regulated by the CaMV 35S promoter (CaMV 35S-Pro). LB and RB indicate left and right T-DNA borders, respectively. (b) Genomic DNA PCR analysis of control and transgenic plants. The presence of BrSRS genes was verified by PCR amplification using gene-specific primers. (c) Semi-quantitative RT-PCR for BrSRS genes in transgenic plants. The actin (At3g19780) gene expression level was used as a quantitative control. Lanes: M, molecular size marker; Wt, untransformed wild type; lanes #1 to #3, overexpressing lines 1, 2, and 3; 1-OX, BrSTY1-overexpressing lines; 2a-OX: BrSTY2a-overexpressing lines; 2b-OX, BrSTY2b-overexpressing lines; 5-OX, BrSRS5-overexpressing lines; 7-OX, BrSRS7-overexpressing lines; L-OX, BrLRP1-overexpressing lines. (d) Phenotypes of T1 transgenic Arabidopsis plants transformed with BrSRS genes. Seedlings were grown for 7 days in the same Petri dish, transplanted to soil, and then photographed 20 days later. Wt, untransformed wild type; 1-OX, BrSTY1-overexpressing lines; 2a-OX: BrSTY2a-overexpressing lines; 2b-OX, BrSTY2b-overexpressing lines; 5-OX, BrSRS5-overexpressing lines; 7-OX, BrSRS7-overexpressing lines; L-OX, BrLRP1-overexpressing lines. Bars are 1 cm.
Supplementary Fig. 2 Phenotypes of transgenic Chinese cabbage (B. rapa L. ssp. pekinensis) plants overexpressing BrSRS7. (a) Comparison of soil-grown plants at the same age before flowering. (b) Phenotypes of BrSRS7-overexpressing transgenic Chinese cabbage after flowering.
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