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SUPPORTING INFORMATION LEGEND
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Figure S1. Growth environments and days to heading of Asominori and NIL (ltg1) in
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three different environments. (a) Day length variation in the Nanjing summer (data
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collected from May 2008 to October 2008) (HTE1; Table S1) and in the dry season of
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Lingshui, Hainan (data collected from December 2008 to April 2009) (LTE1) from
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sowing to harvest. (b) Temperature variation in the natural Nanjing summer (HTE1),
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natural Hainan dry season (LTE1) and Hainan greenhouse (HTE2). (c) Days to heading
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(DTH) of Asominori and NIL (ltg1) in these three environments. Data given as means ±
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s.d. (n = 10 plants). ** represent significant difference (P < 0.01) according to Student’s
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t-test.
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Figure S2. Diurnal expression patterns of genes associated with DTH in rice.
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Expression of Hd1, Hd3a and RFT1 by qRT-PCR was analyzed in Asominori and NIL
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(ltg1) plants grown under different photoperiods and temperatures. In all panels, the Y
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axis represents relative expression levels and the mean of each point is based on the
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average of three biological and three technical replicates ± s.d. calculated using the
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relative quantification method. The black bars indicate the dark period, and the white bars
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indicate the light period. The numbers below the bars indicate the hours of RNA
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sampling.
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Figure S3. Germination rate of Asominori and NIL (ltg1) grown under low
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temperature conditions. Seeds were sawn in a growth chamber under a 12 h day
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photoperiod and at constant temperature of 23ºC. Germination was scored every 12 h
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when root and/or shoot emerged. Each data point represents mean ± s.d. (n = 30).
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Figure S4. Variation in shoot heights in Asominori and NIL (ltg1) grown under
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different temperature conditions. (a-e) Shoot height variation of plants grown at 33ºC
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(a), 30ºC (b), 27ºC (c), 24 (d) and 21ºC (e), all at 12 h day photoperiod. Plant height was
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measured at five growth points in each condition. Data are given as mean ± s.d. (n = 10
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plants).
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Figure S5. Alignment of LTG1 with homologous proteins from Arabidopsis, rice,
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maize and human. Homologous sequences were identified from a Blast search using the
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TGI database (http://compbio.dfci.harvard.edu/tgi). The sequences with highest homologies
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were aligned with LTG1 using CLUSTALW software. Roman numerals refer to protein
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kinase motifs I through XI (Hanks et al., 1988). CKI signature sequences are underlined
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(Graves et al., 1993). The near-consensus SV40T antigen putative NLS is boxed.
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Figure S6. Subcellular localization of LTG1. (a) Localization of GFP, (b) Localization
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of the LTG1-GFP fusion protein in rice protoplasts. Bars = 7.5m.
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Figure S7. Phylogenetic tree of LTG1 and homologous sequences. The tree was
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derived from alignment of full-length amino acid sequences of LTG1 and its homologous
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genes from Arabidopsis, rice, maize and human using ClustalW and the neighbor-joining
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method. The number at each node represents the bootstrap support (percentage) and the
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scale bar indicates the genetic distance based on branch length.
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Figure S8. Expression pattern of LTG1. (a) Analysis of LTG1 expression in roots (R),
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culms (C), sheaths (S), blades (B) and panicles (P) by qRT-PCR. (b) GUS staining of the
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same tissues; bars = 1 mm.
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Figure S9. Expression of LTG1 in plants grown under different temperatures. (a-d)
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Transcript levels of LTG1 in 4-week-old Asominori and NIL(ltg1) plants grown at
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constant 32ºC (a), 23ºC (b). Four-week-old plants grown at 32ºC were transferred to 23ºC
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for 0-72 h (c); plants grown at 23ºC for four weeks were transferred to 32ºC (0-72 h) (d).
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All conditions were with a 12 h photoperiod. The black bars indicate the dark period.
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Figure S10. Effect of auxin treatment on plant growth in Asominori grown under
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LT. (a) Phenotype of germinated Asominori seedlings treated with 0, 1, 10 M NAA for
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two weeks and grown in a hydroponic system and at constant 23ºC. (b) Shoot and root
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lengths of the plants. Bars represent means ± s.d. (n = 18 plants). Different letters indicate
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significant differences according to Tukey’s test at P < 0.05.
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Figure S11. Alignment of the deduced amino acid sequences of the LTG1
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haplotypes. Sequences were deduced from the coding cDNAs of 159 cultivars and
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aligned using ClustalW software. Shaded residues represent positions of those amino acid
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substitutions marked in red compared to haplotype LTG1-H1. Numbers in parentheses
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represent the number of cultivars with each haplotype.
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Figure S12. Phylogenetic relationship of the genomic region surrounding the FNP of
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LTG1. The analysis was done with 45 cultivars and 30 wild races using ClustalW and the
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neighbor-joining method. Green circles and red diamond represent the cultivars and wild
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race respectively, which are of the LTG1-H2 haplotype. The purple circle represents the
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LTG1-H4 haplotype. The scale bar indicates the genetic distance based on branch length.
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Table S1. Characteristics of the seven environments used in this study.
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Table S2. Temperature sensitivity, haplotype, local origin and days to heading
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(DTH) of all cultivars and wild races used in the study.
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Table S3. Molecular marker primers used in map-based cloning of LTG1.
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Table S4. Primers used in sequencing of the final mapping region of LTG1 between
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the markers L264 and IND14.
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Table S5. Primer sequences used to generate DNA constructs.
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Table S6. Primer sequences used in qRT-PCR analysis.