Figure S1. The coding sequence alignment among OsAHPs and OsPHPs, and the relative expression of OsAHPs and OsPHPs in OsAHPs-RNAi and wild-type rice seedlings.

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Presentation transcript:

Figure S1. The coding sequence alignment among OsAHPs and OsPHPs, and the relative expression of OsAHPs and OsPHPs in OsAHPs-RNAi and wild-type rice seedlings. (A) Schematic representation of the OsAHP-RNAi construct. The numbers indicate the nucleotide positions in the OsAHP2 coding sequence. Pubi, maize Ubiquitin promoter. (B) The coding sequences of the genes were aligned; underlining indicates the RNAi fragment used to knock down OsAHP1-2. (C) Measurement of the mRNA levels of two OsAHPs and three OsPHPs in shoots and roots by qRT-PCR.

Figure S2. Expression patterns of OsAHP1 and OsAHP2. (A) Real-time PCR analysis of OsAHP1 and OsAHP2 expression in different tissues. (B-H) GUS staining assay of Pro OsAHP2 ::GUS transgenic rice plants. (B) Five-day-old seedlings. (C) Five-day-old seminal roots. (D) Eight-day-old seminal roots and lateral roots. (E) Lamina joint. (F) Mature leaf. (G) Basal node. (H) Spikelet.

Figure S3. OsAHP2 overexpression partially rescued the Arabidopsis ahp1,2,3,4,5 mutant phenotype. (A) Nine-day-old seedlings of wild-type, the ahp1,2,3,4,5 quintuple mutant, and ahp1,2,3,4,5 mutant expressing the OsAHP2-GFP fusion protein grown under long-day conditions. (B) Confocal image of the OsAHP2-GFP fusion protein in root cells of the transgenic plant shown in (A). (C) Western blot analysis of OsAHP2-GFP expression in two lines of seedlings shown in (A) using anti-GFP antibodies. Rubisco (RBC) was used as a loading reference.

Supplemental Table S1. Primers used for gene cloning. PurposePrimer nameSequence (5' to 3') OsAHPs-RNAi AHPRNAiFGGGGTACCACTAGTACCAGCTCACCGCTCTCCTCT AHPRNAiRCGGGATCCGAGCTCGGGATCATAAGCCTGGATTTGC 2×35S::OsAHP1-GFP OXAHP1FGCTCTAGATCCATTCCATCCATCTCTCTC OXAHP1RGGGGTACCCCATGTTTAGGGTAACAAGCTTGG 2×35S::OsAHP2-GFP OXAHP2FGCTCTAGACTCTTCTCTTCTCTTCTGCCT OXAHP2RGGGGTACCCCTTGCTGCTTGGGATCATAAGC Pro OsAHP2 ::GUS PAHP2FCGGGATCCGGGAGGCACTGCACTATTTC PAHP2RCGGAATTCGGGCGAGCGCAATCGGAAT

GenePrimer nameSequence (5' to 3') OsActin1 ACTIN-FTGTATGCCAGTGGTCGTACCA ACTIN-RCCAGCAAGGTCGAGACGAA OsAHP1 OsAHP1-FCAGAGATGGGTGCCTCAAGAC OsAHP1-RCGCGCCAGTTTAATGTTTAGG OsAHP2 OsAHP2-FGCGCAACAAGTTTCAGACTATGC OsAHP2-RGATCACATGCAGCTACACTCTTTTG OsPHP1 OsPHP1-FAGCTGCTAAGACAAGCTGGC OsPHP1-RTCCTATTTCCAGCTGCTAAGAC OsPHP2 OsPHP2-FCAGTTACTGAGGCAAGTTGGTC OsPHP2-RTACTGTTCTTCTTACTTCCTTGAGC OsPHP3 OsPHP3-FCTGTTGCGACAAGCTGGTC OsPHP3-RCAGCCCGAGACCAATTCAATAG OsRR1 OsRR1-FAGGATCAGCAGATGCATGAATG OsRR1-RGAGACGCTGTACGTCCTTGCTT OsRR2 OsRR2-FGACTAGCCATGGTGATGAATGC OsRR2-RGCTGCCATTGGACCATCTGT OsRR4 OsRR4-FTGGCATGACAGGGTATGATCTG OsRR4-RTCCTTGCAGGCACATTCTCA OsRR6 OsRR6-FGTCCCCAACGTCAACATGATC OsRR6-RCACGTTCTCCGACGACATGAT OsRR7 OsRR7-FTGCTCAAGAAGATCAAGGAATCG OsRR7-RGGCACGTTCTCTGACGACATTAT OsRR9/10 OsRR9/10-FTCATGAGGACAGCCCAATTTCTA OsRR9/10-RTGCAGTAGTCTGTGATGATCAGGTT OsSOS1 OsSOS1-FGAAGAGGTGATTGTCAGAGTCG OsSOS1-RGAGTATTTCCATTGGCTGGTCC OsHKT1;1 OsHKT1;1-FGGCTATGGTAAAAGCTGAAGC OsHKT1;1-RCACTATGGCCCAATTAGAAACC OsHKT1;5 OsHKT1;5-FTTCCATGCACACCCATTCTG OsHKT1;5-RGAAACCCAAGAGGGAGATGAAGA OsNHX1 OsNHX1-FGGCTGATGAGAGGAGCTGTGT OsNHX1-RATTGCCGTGCAGCTGAGTATG OsNHX5 OsNHX5-FATGATGATGTCTTCGGTGAACAA OsNHX5-RTCAGGTGGCAACTCATCCAA OsDREB1A OsDREB1A-FGGAATCAGGAGCAAGCAGAAA OsDREB1A-RCGACTCGCCGCTCATCTC OsDREB1C OsDREB1C-FCGAGTACCAGAGCTGGCAGAT OsDREB1C-RTGTATAGGAGGAGCAAAGCTGGTT OsDREB2A OsDREB2A-FAAAAGCGATGGCCCTGATTC OsDREB2A-RTTGGCTGGCGCTTTCCT Supplemental Table S2. Primers used for qRT-CR analysis.

Coin Figure: Thirty-day-old wild-type (WT) and OsAHPs-RNAi seedlings grown on 250 mM mannitol-containing mediums