Phytoremediation of triphenylmethane dyes by over-expressing a
Citrobacter sp. triphenylmethane reductase in transgenic Arabidopsis
Xiao-Yan Fu, Wei Zhao, Ai-Sheng Xiong, Yong-Sheng Tian, Bo Zhu, Ri-He Peng*,
Quan-Hong Yao*
Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of
Agricultural Sciences, Shanghai, China
*Corresponding authors:
Dr. Quanhong Yao
Director and Professor
Shanghai Key Laboratory of Agricultural Genetics and Breeding
Biotechnology Research Institute
Shanghai Academy of Agricultural Sciences
2901 Beidi Road, Shanghai, People’s Republic of China
Tel.: +86-21-62203180; Fax: +86-21-62203180
Email address: yaoquanhong_sh @yahoo.com.cn
Dr. Ri-He Peng,
Professor
Shanghai Key Laboratory of Agricultural Genetics and Breeding
Biotechnology Research Institute
Shanghai Academy of Agricultural Sciences
2901 Beidi Rd
Shanghai, China
Tel: 86 21 62203180
Fax: 86 21 62205704
Email: pengrihe69 @.yahoo.com
Xiao-Yan Fu and Wei Zhao contributed equally to this article
Supplemental materials：
Table S1 Primers used in this study to synthesize the tmr gene
No.
Oligonucleotide
P1
ATGGCTATCGCTGTCACTGGTGCTACTGGTCAACTCGGTGGTCTTGTCATCCAACACTTTGCT
P2
TTACGAACGATGGCAATGATCTGAGAGGCAGGGACCTTCTTCAGCAAAGTGTTGGATGACAAGA
P3
AGATCATTGCCATCGTTCGTAACGTCGAGAAAGCCTTCCACTCTTGCTGATCAAGGTGTCGAA
P4
CTGAAAGAGACTCAGGTTGATTGTAGTCACCATGACGAACTTCGACACCTTGATCAGCAAG
P5
AACCTGAGTCTCTTTCAGAAGGCTTTCGCTGGTGTCTCCAAGCTGCTCTTCATCTCTGGTCCTCA
P6
CGACGTTAGCATGTTGGACGATCAGCAGAGTGTATGTCGTAGTGAGGACCAGAGATGAAGAGCAG
P7
CGTCCAACATGCTAACGTCGTCAAGGCTGCTCGTGATGCTGGTGTCATAGCACATCGCTTACAC
P8
TGAGCAAGTGGAATGATGGATTCCTCAGCGAAAGCGTAACCAGTGTAAGCGATGTGCTATGACA
P9
CCATCATTCCACTTGCTCACGTTCCACCTTGCTACTGAGTACGCTATCCGTACTACCAACATTCC
P10
ACGAAGAAGTCAGTGTACAAAGCGTATACGAAGGAAGGTGTATGGAATGTTGGTAGTACGGATA
P11
TGTACACTGACTTCTTCGTCAACGAAGGTCTGCGTGCTTCCATCGAGTCTGGTGTCTATCG
P12
TCGAGTCTGGTGTCTATCGTCACCAATGCTGGTAGTGGTATCGTCAACTCCGTCACTCGTAAC
P13
TCAACTCCGTCACTCGTAACGAACTTGCTCTTGGCTGCTGCTACTGTTCTGACTGAGGAAGGTC
P14
CCAAGGTTGGTTGGAAGACCAGGTTGTAGGTCTTGTTCTCGTGACCTTCCTCAGTCAGAACA
P15
GGTCTTCCAACCAACCTTGGACCTTCGACGAACTTGCTCAGATCCTCTCTGAGGTCTCTGGC
P16
TTCTCTTCTTCGAAAGAGACAGGCTGATGGACGACCTATCTTGCCAGAGACCTCAGAGAGG
P17
TCTCTTTCGAAGAAGAGAAGAACTTCCTTGTCAACGCTGGTGTTCCCTGAGCCATTCGCTGAG
P18
GAAGACCTCACCTTTGGAGATAGCGTCGTAGATAGCAGCAGTGATCTCAGCGAATGGCTCAGG
P19
CTCCAAAGGTGAGGTCTTCCAAGACCTCTGATGATCTTCAGAAGCTGATCGGTTCCTTGACTCC
P20
ATTACATCTTCAGGGCTTGTTTGACGGATCTCCTTCAGAGGAGTCAAGGAACCGATCAGCT
Fig. S1. The PTDS strategy for the synthesis of the tmr. Oligonucleotides of 60 bp were assembled as
indicated in the graph. There was a 20 bp overlap for each pair of overlapping oligonucleotides.
Fig. S2. Tolerances of transgenic lines (1-2, 5-8, 7-4) in comparison with the wild-type to crystal violet and
malachite green . WT and transgenic T2 seeds were directly germinated and grown for two weeks
vertically on the agar MS plate supplemented with 8.0 mg/L CV (a) and MG (b).
Fig. S3. Tolerance of wild-type (WT) and transgenic lines (5-8, 1-2) to 4.0 mg/L leucomalachite green
(LMG).