Fig. S1 Gene expression profiling of OsSUS3 in life cycle of rice.

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Fig. S1 Gene expression profiling of OsSUS3 in life cycle of rice. The X-axis indicates the 33 developmental tissues with 1-5, calli; 6, seed imbibition; 7, seed germination; 8-11, plumule/radicle under dark/light; 12, seedlings; 13, young shoot; 14, young root; 15-16, mature/old leaf; 17-18, mature/old sheath; 19-20, young/old flag leaf; 21-25, panicle at 5 developmental stage; 26-27, young/old stem; 28, hull; 29, spikelet; 30, stamen; 31-33, endosperm of 7/14/21 DAP (days after pollination). The Y-axis presents the expression values obtained from the microarray analysis.

Fig. S2 Biomass enzymatic saccharification and ethanol production of the OsSUS3-transgenic rice plants. (a) Hexose yields released from enzymatic hydrolysis after the pretreatment with 1% NaOH or 1% H2SO4. (b) Bioethanol yields obtained from yeast fermentation using the sugars released from biomass enzymatic hydrolysis as performed in (a). a b All data are given as means ± SD. A Student’s t-test was performed between transgenic plants and ZH11 as **P < 0.01 and *P < 0.05 (n = 3).

Table S1. Biomass and seed yields in two-year field experiments. Transgenic line Biomass production (g/plant) Seed yield (g/plant) 2013 2015 ZH11 31.81±0.96 24.73±1.29 34.01±1.69 23.92±1.15 Vector EV 31.63±2.18 23.92±0.63 33.56±2.24 23.92±1.65 AtCesA8 ::SUS3 1 34.67±1.75* 26.52±1.11* 37.19±3.94 25.12±0.82 2 35.30±1.11* 26.82±1.01* 37.20±3.54 25.62±0.92* 3 34.50±1.73* 26.28±0.99* 36.36±2.81 25.50±0.39* 4 33.52±0.35* 26.55±1.25* 35.54±1.92 25.90±1.02* * and **, Indicated significant difference between transgenic lines and ZH11 control by t-test as P < 0.05 and 0.01 (n=5)

Table S2. Three major wall polymer levels and CrI values in the mature stem tissues. Cellulose content (%dry matter) Hemicellulose content Lignin content CrI (%) ZH11 22.35±0.13 16.34±0.10 12.85±0.17 52.55±0.61 Vector CK 22.62±0.44 16.56±0.10 12.79±0.95 51.51±0.66 AtCesA8 ::SUS3 1 25.15±0.15** +13%@ 18.74±0.19** +15% 13.90±0.97 48.62±0.35** -7% 2 24.45±0.12** +9% 17.89±0.12** 13.42±0.63 49.30±0.58** 3 26.13±0.29** +17% 19.79±0.07** +21% 13.26±1.16 47.40±0.64** -10% 4 23.14±0.24* +4% 19.80±0.07** 12.84±0.52 47.74±0.53** -9% * and **, Indicated significant difference between transgenic lines and ZH11 control by t-test as P < 0.05 and 0.01 (n=3); @ Percentage of increased or decreased level between transgenic line and ZH11 by subtraction of two values divided by ZH11.

Table S3. Sucrose, fructose and glucose contents of OsSUS3-transgenic rice plants in mature stem. Transgenic line Sucrose (µg/g FW) Fructose (µg/g FW) Glucose (µg/g FW) ZH11 1019.47±55.42 78.19±0.76 43.70±1.19 Vector EV 941.28±28.93 74.56±0.23 41.92±0.36 AtCesA8:: SUS3 1 571.47±19.44** -44%† 74.98±1.32 39.51±1.11** -10% 2 392.13±7.40** -62% 37.70±0.90** -52% 17.10±0.42** -61% 3 410.93±10.51** -60% 63.84±0.12** -18% 31.61±0.95** -28% 4 459.61±7.28** -55% 41.87±0.77** -46% 17.83±0.48** -59% FW: fresh weight; * and ** Indicated significant difference between transgenic line and ZH11 by Student’s t-test at P < 0.05 and P < 0.01; † Percentage of increased or decreased level between transgenic plants and ZH11 by subtraction of two values divided by value of ZH11.

Table S4. Primers used for Q-PCR analysis. Gene Purpose Forward primers Reverse primers AtCesA8 Promoter clone CCCAAGCTTCAGAGGAAACTCAGATGTGATGA ACGCGTCGACCTTCGAATTCCCCTGTTTGGAGA OsSUS3 Gene clone TCCCCCGGGATGGGGGAAACTACTGGAG TCCCCCGGGCATTGCCCATTCTTTTCAC Q-PCR GTGAGACACGCCGCTACCTGGAGA CCCTGAAATCAACTAAACCCT OsCESA1 GGCATGGTGGCGGGTATAT AGGCGACGGCTTTCTGAGT OsCESA3 GTGTAGATAGAAACACGAAAGGTG CAATGGGCACCCACACACGT OsCESA4 CCATCGTCTACCCGTTCACC TCCAGCCCAGCAATCATCTT OsCESA7 TTAACGGGGTTCAAGATGC ATGGTGTTGGTGTAGGCGA OsCESA8 TGGTCGGTGTTGTTGCTGG ATGGTCGGAGTGCGGTTTT OsCESA9 TACAAGAACGGCAACCTCA AAGAACAAACTCGCAAACG OsGH9B1 ACTTCCTCTACAAAACCCCAAC ATAATCCACTCGCTGAACCATA OsGH9B3 TCCGATCTCCTCTACGACTGA CCCCGTTCCTGCCATTT OsGH9B16 CGGAACCACCAAGTAGCCAT GAATACACGACAAAGTCAACGAAC OsIRX9 ACAAGGAGATGATGATTTTAGCA AGGCGGAGGTCGTAGATGTCG OsIRX14 GATGGATGGTGTGATTGTG TTGAACTGGAAGAGGCATA OsUBQ1 CCAGGACAAGATGATCTGCC AAGAAGCTGAAGCATCCAGC