From the Cover: AtALMT1, which encodes a malate transporter, is identified as one of several genes critical for aluminum tolerance in Arabidopsis -- Hoekenga et al. 103 (25): 9738 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Hoekenga et al. 10.1073/pnas.0602868103.

Supporting Information

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Supporting Figure 8
Supporting Figure 9
Supporting Table 2
Supporting Table 3

Fig. 8.

Ecotype survey of Al tolerance. Al tolerance was estimated for nine ecotypes plus AtALMT1 MT by using root growth (vertical axis) and Al-activated malate efflux (horizontal axis). Median values are reported; error bars indicate standard error values. Superscript numbers indicate the five distinct Al tolerance classes according to ANOVA of root growth. The correlation between root growth and malate efflux is r = 0.71 (black line) for all of the varieties tested.

Supporting Figure 9

Fig. 9.

Examples of families of currents recorded in the absence (-) and presence (+) of Al³⁺ from AtALMT1-expressing and control cells preloaded with malate. Currents recorded in AtALMT1-expressing cells but not preloaded with malate are shown in Center. The holding potential was set to 0 mV, and voltage pulses were stepped between +10 and -200 mV (in 10-mV increments). For clarity, only currents in response to 20-mV steps are shown.

Table 2. Molecular genetic markers designed for this study

Name	Position on chr 1, kb	Forward primer sequence	Reverse primer sequence	Note
T25K16	37.3	TCCCCGGAGAAGTCATCGTT	GGCTTCTCTTCTTCTTCTTCCAGTCT	Col 189, Ler 180
F10O3	746.9	TTAATGAAACCAACCCAAGAT	TGCGAAGAACCACTAAACC	Col 292, Ler 244
F21B7-1334	923.9	TGACTACATGGAGATTATGGCC	CACGATATGATCAAGCTTTAACG	From arabidopsis.org
F21M11	1,022	CTTCTCTGCGAACGTTACTCTTCTTC	CGCCGATACAAACTCGCCATAG	Col 195, Ler 185*
F20D22	1,070	AACAAAATGAGTTTCTCTGCATG	CCCAAGTGACGTCTGGTTTC	Col 179, Ler 171
AT1G4140	1,079	GCCGATGTGGATGCTGATTC	TGGAGGAGGTACAAGAGGACA	MboI cuts Ler
F19F19-S	1,158	CCACGTAGGTCAAGAAGAAGAAG	TGTCTGCTGCGATAGAGAGAG	Col 205, Ler 195
F19P19-C	1,198	AGCAATAAGCCGCTTCTTTAGG	AGTGCCAGTGGCTTTTTACCA	HpyCH4V cuts Ler
T1G11	1,243.3	GAAGACAAAGCTCTGCAGTAATGTAG	TTTAGATTTTTGCGTATTGAACAGAT	Col 237, Ler 216
T7A14	1,471.5	GATTCGCATTTCCATTTAGTTTTT	ATGTGTTACTTAAGCCCGTTTTT	Col 310, Ler 249
YUP8H12	1,520.2	TGAGACTTACGAGGATGAAACAAACT	TGCGATAGGGTGGCGAATAG	Col 300, Ler 270
F3F20	1,691	CATCGATGAAGGTGAGAAGTGT	ATATACCCATGCATCAAATCAAA	Col 140, Ler 128
F12K11	1,997.3	TTGTATAAAGAAGAGCCTGTCACC	TTCTCGAATCTAACCAAATGTCTT	Col 223, Ler 179
F4H5	2,105	AGAGATGAATGCTTTGGTTGTGT	CTACGCTCTTCATTTCATTTCATTTG	Col 157, Ler 145
F24B9-2	2,368.8	CTTTCTTTCTGTTGTATTATCGTAGC	ATGTCCTGGGTAGTCACTTCCTTTAT	Col 156, Ler 135
F24B9-1	2,429	AGCTCTCGTCGCCGTCGTCTCTAC	CTGCGTGTGGCTGTTTTGCTTTTGT	Col 112, Ler 102
T23G18	2,573.4	GTTTCTCTTCCTTCTCTTTA	CCATACCTGAAACATTTACA	Col 195, Ler 189
T27G7-1	2,630.5	ATGGGCCACATCCTTATTTGGTT	CTTATCCTTTGAGGCTTTCAGTTTGA	Col 161, Ler 151
T27G7-2	2,660	GTGCCTACAGGATTGATGC	ACCTGATGTTATTTTTGTGTTT	Col 238, Ler 218
T27G7-3	2,669	TTTGGGGATTCTCATTAGTTTTTC	AGCACCCAGACAGTTACCTTC	HindIII cuts Col
F22O13	2,703.1	TGAGCGATTTAAGGTTATCT	TTTCTTTTCTTTTCTTTTCT	Col 158, Ler 148
F7G19	2,890.9	CAAAAGTGTTAACGATGTAGAGG	ATTGTGGATTATGAAGGAAAAAC	Col 201, Ler 178
T10O24	3,487.3	CTACGCCATCACCTGCCATTT	TCCTTACATTTATTCTCTGCCAACAC	Col 297, Ler 269*

*Markers used for initial recombinant screening.

Marker occurs within the fifth intron of AtALMT1.

Table 3. Primers used for cloning, sequencing, and hybridization

Name	Sequence	Position on chromosome 1, bp
Northern-F	CATGAAAAACATGAAGATGGTGA	2660966988
Northern-R	CGCCGGTCTATGTTCT	2661341326
Cloning-F	TGTTGAGGGCACTAATCTTGTCG	2658269291
Cloning-R	GCGCCGGTCTATGTTCTTTTATTTACT	2661342316
Sequencing-1	ACACTTGAGAGAGCTGAGTG	2658763783
Sequencing-2	GTGGAACCTATTCTTTTGGT	2659233252
Sequencing-3	TCATTATGTCTCTACAAGAA	2659709728
Sequencing-4	TATTTACATGTCGCAAATGC	2660273292
Sequencing-5	TGCTATACGAGAAGTCGGAT	2660772791