SUPPLEMENTARYINFORMATION

Callus culture and regeneration

Rice calli were induced from immature embryos (12-15 days after pollination) of a japonica variety Zhonghua 11 cultured on callus induction medium. The vigorous granular calli were selected for pre-differentiation culture and 15 days later for differentiation culture. When the regenerated shoots reached to about 1 cm in height, they were transferred for rooting culture. The media used in different stages were as follows:

Medium for callus induction and subculture:N6 (Chu 1978) macro nutrients, B5 (Gamborg et al. 1968) micro nutrients and vitamins, 2,4-D 2 mg/L, casein hydrolysate 300 mg/L, L-proline 500 mg/L, L-glutamine 500 mg/L, sucrose 30 g/L and agar 8 g/L (pH 5.8);

Medium for pre-differentiation (Yang et al. 1999): N6 macro nutrients, MS micro nutrients (Murashige and Skoog 1962), B5 vitamins, CuSO4·5H2O 1 mg/L, ABA 5 mg/L, 6-BA 6 mg/L, NAA 2 mg/L, casein hydrolysate 500 mg/L, L-proline 500 mg/L, sucrose 30g/L and agar 8 g/L (pH 5.8);

Medium for differentiation: N6macro nutrients, MS micro nutrients, B5 vitamins, 6-BA 3 mg/L, NAA 1 mg/L, sorbitol 18 g/L, sucrose 20 g/L and agar 8 g/L (pH 5.8); Medium for rooting: N6 macro nutrients, MS micro nutrients, B5 vitamins, NAA 1 mg/L, sucrose 20 g/L and agar 8 g/L (pH 5.8).

SUPPLEMENTARY FIGURE

Fig. S1Expression patterns of wild-type candidate genes (a: LOC_Os01g60730; b: LOC_Os01g60740) in the Psd1 locus, based on the Rice Expression Profile Microarray Database (

SUPPLEMENTARY TABLES

Table S1Plant height of ZH11 and Psd1 plants grown in different seasons

Population / Growing seasona / Monthly mean
day length (h) / Plant height (cm)
Psd1 / May, June, July / 13.2, 13.5, 13.4 / 24.7 ± 2.4
Aug, Sept, Oct / 12.9, 12.3, 11.6 / 58.3 ± 3.2
ZH11 / May, June, July / 13.2, 13.5, 13.4 / 88.4 ± 4.1
Aug, Sept, Oct / 12.9, 12.3, 11.6 / 81.5 ± 3.7

a The plants were grown in the field in Guangzhou in middle May (or August)2000 and the major tillers headed in late July (or October).Plant heights are means ± SD, N = 15.

Table S2Primer sequences of PCR-based markers

Marker (Type) / Forward (5' to 3') / Reverse (5' to 3')
RM302 (SSR) / TCATGTCATCTACCATCACAC / ATGGAGAAGATGGAATACTTGC
RM486 (SSR) / CCCCCCTCTCTCTCTCTCTC / TAGCCACATCAACAGCTTGC
135250 (InDel) / GAAGCATTTCTTTGGTGCAAAC / GCATCCGAGGACAAAAAACAC
135680 (InDel) / CGTGCTAGTTTGGATTGAAGG / CGTGGTGCCTTTTTTCGAGCTG
136730 (InDel) / TTCAACTCCAGGAGGAACTCC / CATGCACGAGAATAAAGCGGCA
135136 (InDel) / CGAAACTGGACAAAGAGTATG / GCGTAGAGCGTACACGAGTC
35.087 (SNP) / TAGCGACTTGCGAGAAGCCAT / GCCTCATCTTTTCCCTTATGC
35.087ZH-R / GATCACTAAGAACACTTAT
35.087Du-R / GATCACTAAGAACACTTAC
35.112 (SNP) / CGTAGTCGACAACGCAGTAGT / GGCCCACCAAGTACCAAGTCT
35.112ZH-R / CGGATTGTATCCATCTTA
35.112Du-R / CGGATTGTATCCATCTTG
35.124 (SNP) / CGCGAGCTGGTAGCAGATGG / GCCGTGTCGTGTAAAAACTTG
35.124ZH-R / ATGACGTGTAATTCGTTG
35.124Du-R / ATGACGTGTAATTCGTTT
35.191 (SNP) / CCGCTCACTTGGACGAACTC / ACGAGACTATTCGGATTTAGG
35.191ZH-R / CCTCAAACATGGATCTG
35.191Du-R / CCTCAAACATGGATCTA
ORF2-3' (HRM) / CTGCCTCAAGAACGTCGCCAA / CTTGTAAGGGAGGGAGACGTT

Note: 35.087, 35.112, 35.124 and 35.191 are SNP markers by semi-nested PCR; the primers with “-R” are allele-specific primers, and the polymorphic bases are underlined.ORF2-3' marker is based on high-resolution melting (HRM) analysis.

Table S3Primer sequences used for qRT-PCR analysis

Primer / Forward (5' to 3') / Reverse (5' to 3')
Actin1 / CTCAGCACATTCCAGCAGAT / ACAGATAGGCCGGTTGAAAA
qORF1 / GCTCGTCTGTTCAGTTTGTAGG / CTGTACGTAACTGTAAGCGTCC
qORF2 / GGGATCGTATGTCTCCACAGTT / ATGGACGGTTGACACAGTTCAC
qDTH8 / CAGCGCCGGGTATGTCGTCT / GTCGTCGCCGTTGATGGTCTT
qGhd7 / ATATTGTGGGAGCACGTT / ATCTGAACCATTGTCCAAGC
qRFT1 / TACTTCAACTGCCAGCGCGAGG / AGCTATAGCTGCTGCATGCATGGA
qEhd1 / GCGCTTTTGATTTCCTGC / TTCGGAATATGTGCTGCC
qHd3a / GCTCACTATCATCATCCAGCATG / CCTTGCTCAGCTATTTAATTGCATAA

SUPPLEMENTARY REFERENCES

Chu CC (1978) The N6 medium and its applications to anther culture of cereal crops. Proceedings of the Symposium on Plant TissueCulture Held at Peking, China during 25–30 May. Science Press,China. pp. 45–50

Gamborg OL, Miller RA, Ojima O (1968) Nutrient requirements of suspension cultures of soybean root cell. Exp. Cell Res.50:151–158

Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 39:375–383

Yang Y, Zheng Y, Chen Y, Jian Y (1999) Improvement of plant regeneration from long-term cultured calluses of Taipei 309, a model rice variety in in vitro studies. Plant Cell Tiss.Organ Cult. 57:199