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Supplementary Material

Table S1. Primers used for sequencing and PCR analysis.

Primer1 / Species / Sequence (5’to 3’) / Position on cDNA F. linearissequence.
FZ-1(F) / spinach / gtcacattgttcttggct / 93 to 113
FZ-3(R) / spinach / cat ccgctaccatacttcc / 726 to 744
3’race(F) / F.linearis / tgttaagcactg gat ctggtgtgaatg / 511 to 537
5’race(R) / F.linearis / ccattcacacca gat ccagtgcttaac / 512 to 538
FBP-1(F) / F.linearis / gctgtgctgctgttataaggc / 119 to 142
FBP-1(R) / F.linearis / ctt tgg aat ctt aat gtc ag / 583 to 602
FBP-2(R) / F.linearis / ccc taa aga tgg atc aag ag / 544 to 563
FBP-3(F) / F.linearis / gat ggaacaagctggagg ac / 851 to 868
FBP-4(R) / F.linearis / ctcttctccctg cac att ag / 178 to 195
FBP-5(R) / F.linearis / gccaaaccagccttataacag c / 129 to 142
FBP-6(F) / spinach UTR / gcaatgcgtacggacttgatg / -1 to 12bp
FBP-7(R) / F.linearis / ggaaataca act gcacaagaa c / 1014 to 1034
FBP-8(R) / F.linearis / ggaaag act tcatacaga ac / 819 to 838
PPDK-1(F) / F.brownii / tcagggagaggatgttgttg
PPDK-1(R) / F.brownii / ctctttccttgtgcatgcc

1 F and R indicate the forward and reverse sequences in relationship to the F. lineariscytosolic FBPase sequence.

Table S2. Sequence length and overlap.

band size / Primers used / Where on sequence
200bp / FBP-6(F)/FBP-4(R) / -1-178bp
550bp / FBP-6(F)/5’Race / -1-513bp
650bp / FZ-1/FZ-3 / 113-726bp
500bp / FBP-1(F)/FBP-1(R) / 142-582bp
700bp / 3’Race/pcr anchor primer (R) / 537-1120bp
1020bp / FBP-1(F)/ anchor primer (R) / 142-1120bp

Table S3. Coding and abbreviations used for Fig. 11.

code / Abbreviation / Arabidopsis mutants / Name
1 / cytFBPase / cytosolic fructose 1,6-bisphosphatase
2 / PFP / pyrophosphate-dependent phosphofructokinase
3 / PFK / ATP-dependent phosphofructokinase
4 / SPS / sucrose phosphate synthase
5 / ADPase / ADP-glucose pyrophosphorylase
6 / GWD/GWD1 / sex1 / α-glucan, water dikinase
6 / PWD/GWD3 / phosphoglucan, water dikinase
7 / ISA / isa1, isa2, isa3 / Isoamylase (three isoforms, with ISA3 being main active form)
8 / BMY9 / β-amylase (there are four isoforms)
9 / DPE1 / dpe1 / D enzyme or disproprtionating enzyme or α-1,4-glucan transferase
10 / DPE2 / dpe2 / transglucosidase or amylmaltase
11 / PHS2 / α-glucan phosphorylase (cytosolic)
12 / HXK / hexokinase
13 / AMY3 / α-amylase
14 / PHS1 / ph1 / glucan phosphorylase (chloroplastic)
15 / LDA / lda / limit dextrinase

FigureS1. CytFBPase activity (nmol FBP g-1 fresh weight min-1) for the wildtype line 85-1 over the 5 month period of the F2-selfed populations when assayed at room temperature (21-24oC). The conversion factor to express the data as μmolFBP m-2 leaf area s-2 is 0.0103. Values are the mean±SD (n=12-32). These values were used as a reference for the determinations shown in Fig. 2. The activities do not show any statistical difference over time but there is a trend upward in October and November. The room temperature tends to increase in these months, which could cause activities to rise.

Observations on Seed Germination and Crosses

Continuous light significantly increased F. linearisseed germination in petri plates compared to darkness (data not shown). The F1 progeny of a cross between the mutant 84-9 (maternal) to wildtype85-1 cross consists of four plants, all of which had intermediate cytFBPase levels (Micallef and Sharkey, 1996). The F2-selfed progeny gave a combined percent seed set and germination of 3-28%, which was lower than that for F2s from reciprocal crosses (data not shown). The use of F2s from a selfedF1 is a more accurate model for the analysis of the inheritance of the low cytFBPase gene than F2s from a reciprocally crossed F1, because it allows for unknown background genes to be expressed and removed from the model. The F2-selfed progeny gave a day-neutral to normal flowering trait (DN: N) ratio of approximately 1:3, χ2 5.991, df=1, α=0.05,