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Supplemental Figure 1

GhCWIN1 (1) Atßfruct2 (1) Lin6 (1) NtCWIN1 (1) OsCWIN1 (1) ZmCWIN1 (1) Atßfruct4 (1) GhVIN1 (1). GhCWIN1 (45) Atßfruct2 (45) Lin6 (50) NtCWIN1 (49) OsCWIN1 (47) ZmCWIN1 (52) Atßfruct4 (98) GhVIN1 (96). GhCWIN1 (130) Atßfruct2 (129) Lin6 (133)

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Supplemental Figure 1

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  1. GhCWIN1 (1) Atßfruct2 (1) Lin6 (1) NtCWIN1 (1) OsCWIN1 (1) ZmCWIN1 (1) Atßfruct4 (1) GhVIN1 (1) GhCWIN1 (45) Atßfruct2 (45) Lin6 (50) NtCWIN1 (49) OsCWIN1 (47) ZmCWIN1 (52) Atßfruct4 (98) GhVIN1 (96) GhCWIN1 (130) Atßfruct2 (129) Lin6 (133) NtCWIN1 (132) OsCWIN1 (130) ZmCWIN1 (135) Atßfruct1 (198) GhVIN1 (190) GhCWIN1 (224) Atßfruct2 (224) Lin6 (229) NtCWIN1 (227) OsCWIN1 (226) ZmCWIN1 (232) Atßfruct1 (292) GhVIN1 (285) GhCWIN1 (317) Atßfruct2 (320) Lin6 (324) NtCWIN1 (322) OsCWIN1 (318) ZmCWIN1 (326) Atßfruct1 (389) GhVIN1 (380) GhCWIN1 (414) Atßfruct2 (417) Lin6 (421) NtCWIN1 (419) OsCWIN1 (415) ZmCWIN1 (426) Atßfruct1 (487) GhVIN1 (479) GhCWIN1 (499) Atßfruct2 (513) Lin6 (510) NtCWIN1 (508) OsCWIN1 (503) ZmCWIN1 (515) Atßfruct1 (575) GhVIN1 (558) Supplemental Figure 1

  2. Supplemental Figure 1. Alignment of GhCWIN1 sequence with five plant CWINs (Atβfruct2, Lin6, NtCWIN1, OsCWIN1 and ZmCWIN1)and two VINs (Atβfruct4 and GhVIN1). Thirteen boxed sequences indicate the conserved regions of plant invertases. The β-fructosidase motif (NDPD/NG) and cysteine catalytic domain (WECP/VD) were underlined. Arrowsheadsdenote five consistently different amino acids between VINs and CWINs. Arrow show GhCWIN1 signal sequence cleavage site between residue 19 and residue 20.

  3. Plant CIN Plant VIN Bacterial invertase Yeast invertase Plant CWIN 0.1 Supplemental Figure 2 Supplemental Figure 2. Phylogenetic analysis classifies GhCWIN1 as a plant CWIN. Genbank accession number for each invetrase listed and the corresponding species were listed in Supplemental Table 1.

  4. Seed Filial tissue 5 10 15 20 DAA GhCWIN1 GhINH1 GhMST1 GhSUT1 Gh18srRNA Supplemental Figure 3 Supplemental Figure 3. Transcript level of cotton cell wall invertase GhCWIN1,invertase inhibitor GhINH1, monosaccharide tranporterGhMST1, sucrose transporter GhSUT1 in cotton seed and filial tissueat different development stages. Gh18srRNAwas used as an internal control.

  5. A a a 25 20 15 10 5 0 25 20 15 10 5 0 b b b b c c Glucose content (mg g-1 FW) Glucose content (mg g-1 FW) 5 10 15 20 5 10 15 20 Seed Seed coat Seed Filial tissue B 25 20 15 10 5 0 25 20 15 10 5 0 a a ab b b bc b c Fructose content (mg g-1 FW) Fructose content (mg g-1 FW) 5 10 15 20 5 10 15 20 Seed Seed coat Seed Filial tissue C 25 20 15 10 5 0 25 20 15 10 5 0 a ab bc Sucrose content (mg g-1 FW) Sucrose content (mg g-1 FW) c Supplemental Figure 4 Supplemental Figure 4. Glucose (A), fructose (B) and sucrose (C) level in cotton seed, seed coat and filial tissue during development. Each value is the mean ± SE of at least three biological replicates. Different letters indicate significant differences at P≤.05 according to randomization one-way ANOVA test. 5 10 15 20 5 10 15 20 Seed Seed coat Seed Filial tissue

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