Plant Molecular Biology

Are rice (Oryza sativa L.) phosphate transporters regulated similarly by phosphate and arsenate? – a comprehensive study

E. Marie Muehe, Jochen Eisele, Birgit Daus, Andreas Kappler, Klaus Harter, Christina Chaban1

1Centre for Pant Molecular Biology (ZMBP), Plant Physiology, University of Tuebingen, Auf der Morgenstelle 32, 72076 Tuebingen, Germany; E-mail:

Supplementary data

Fig. S1Experimental setups for plant growth response and gene expression experiments.

(a) Glass tanks are filled with microscopic slides holding seeds; the insert shows seeds glued husk-down to a microscopic slide. (b) Germinating seed glued to a microscopic slide. The parentheses indicate the measurement parameters. (c) Plastic boxes containing tubes with germinated seedlings. (d) Plastic tube with a cut-off tip filled with agar and young seedling. (e,f) Schematic representation of the time outline for gene expression experiments.(e) For long-term stress exposure, rice plants were grown for 7days with supplementation of either 10µM or 100µM As(V) or Pi before harvesting. (f) For short-term stress exposure, rice plants were grown for 16hours with 10µM As(V) or Pi before harvesting. Control groups without the amendment of As(V) or extra Pi were setup for each time period as well.The amendment of As(V) and Pi is indicated in red; the harvesting of the plant material is indicated in green. am – morning; pm – afternoon.

Table S1: Primer sequences used for amplification of PT genes by qPCR

Gene / Primer sequence (5’-3’) / *Probe# / Detection
PT1 / ctactggcggatgaagatgcctcctcaatctgggtgtgc / #70 / FAM
PT1 / ctcttctcgcgccagttc
tggaacaggttctggctgta / #77 / FAM
PT2 / cacaaacttcctcggtatgct
gtcgacgacgttgtccttg / #65 / FAM
PT3 / catgctcatgacgctgct
gcgacgttctccttggac / #65 / FAM
PT4 / ggacatcgccttctactcgt
cccgggagatcttgaaca / #65 / FAM
PT5 / cgcctactactcgcagaacc
cccgggagatcttgaaca / #65 / FAM
PT5 / tcgacgagcaggagaagg
aattcccgggagaagagc / #70 / FAM
PT6 / cggagtccaaggggaagt
agcgctacagtacagtttgcag / #65 / FAM
PT7 / gatgacggcgttcatgct
aagaagaaggtgagcgcgta / #70 / FAM
PT8 / aaggccaagaccatgtcg
aagacggtgaaccagtagcc / #65 / FAM
PT8 / ggatgaagatgccggagac
tctcgacctggagcacctt / #70 / FAM
PT9 / gcctggcggatcatactc
ccgtatacctggctgtctcc / #65 / FAM
PT9 / gttcggcatcctcgtcag
gggtagccggtgtagtggt / #70 / FAM
PT10 / ggcggatcattctcatgg
gccgtatatctggccgtct / #65 / FAM
PT10 / ttctccatgcaggggttc
cggggtagccggtgtagt / #70 / FAM
PT11 / tctcaagtcaaaagcatcaagaa
gtgaagaagaagccgagcat / #136 / FAM
PT11 / gagatctccggcgaggac
cttctcgtccctgctcaca / #70 / FAM
PT12 / gggttcctcatgacatggat
tggcagaagcttcctcgt / #65 / FAM
PT13 / caggacgagtatggcctctt
tcgaggacgaaccaacaga / #39/#10 / FAM
* The Universal ProbeLibrary (Roche Applied Science) probes were used.

Table S2: Primer sequences used for amplification of non-PT genes by qPCR

Gene / Primer sequence / *Probe# / Detection/Source
SMT3 / cagctcaagcgtgatccag
tgaccttgaggttgatgtgg / #70 / FAM/Sybr
PHF1 / tttcctcaattgaattttgtcct
cagcagggacattcagcttt / #32 / Sybr Green/{Chen, 2011 #1795}Chen et al. 2011
PHO1;2 / cgcctagcatggactgagagtgt
gttcacggaatggtaatgggaca / Sybr Green/Secco et al. 2010
PHO1;2anti / ggctactagcttggtgctcttctcc
caaggccactatctgtccctgtatg / Sybr Green/Secco et al. 2010
SPX-MFS1 / gtgtttcccttgcgacgtagag
ccgagtcctccgggctaa / Sybr Green/Lin et al. 2010
SPX-MFS2 / acgattcggtggatttgttga
cggcgaaacgatctctcct / Sybr Green/Lin et al. 2010
PHR2 / gaccagaattgtctgaaggttctt
acgcaatgcctcagtgagat / #8 / Sybr Green/Liu et al. 2010
PHO2 / cgagaattttgtcaaggagca
tcacgagcatgtccaacaa / #149 / Sybr Green/Liu et al. 2010
SPX1 / cgacttccatggcgagat
tcctcttgtcatacttcttgagaatc / #132 / Sybr Green/Liu et al. 2010
* The Universal ProbeLibrary (Roche Applied Science) probes

Chen J, Liu Y, Ni J, Wang Y, Bai Y, Shi J, Gan J, Wu Z, Wu P (2011)OsPHF1 regulates the plasma membrane localization of low- and high-affinity inorganic phosphate transporters and determines inorganic phosphate uptake and translocation in rice. Plant Physiol 157(1): 269-278

Lin SI, Santi C, Jobet E, Lacut E, El Kholti N, Karlowski WM, Verdeil JL, Breitler JC, Perin C, Ko SS, Guiderdoni E, Chiou TJ, Echeverria M (2010) Complex regulation of two target genes encoding SPX-MFS proteins by rice miR827 in response to phosphate starvation. Plant Cell Physiol 51(12): 2119-2131

Liu F, Wang ZY, Ren HY, Shen CJ, Li Y, Ling HQ, Wu CY, Lian XM, Wu P (2010) OsSPX1 suppresses the function of OsPHR2 in the regulation of expression of OsPT2 and phosphate homeostasis in shoots of rice. Plant J 62(3): 508-517

Secco D, Baumann A, Poirier Y(2010) Characterization of the rice PHO1 gene family reveals a key role for OsPHO1;2 in phosphate homeostasis and the evolution of a distinct clade in dicotyledons. Plant Physiol 152(3): 1693-1704