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1 Supporting Information Figs S1–S8, Tables S1–S4 1 2 Fig. S1 3 4 Fig. S1 Phenotypes of wild type and wp1 5 A, 20-day-old seedlings of wild type and wp1 6 B, 5-day-old seedlings of wild type and wp1 grown in the dark.. 7 C， PCR detection of wp1. 0.9 kb product could be amplified from positive sample using primers 8 of hygromycin resistant gene 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27

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Page 1: Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 · 1 1 Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 4 5 Fig. S1 Phenotypes of wild type and wp1

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Supporting Information Figs S1–S8, Tables S1–S4 1 2 Fig. S1 3

4 Fig. S1 Phenotypes of wild type and wp1 5 A, 20-day-old seedlings of wild type and wp1 6 B, 5-day-old seedlings of wild type and wp1 grown in the dark.. 7 C，PCR detection of wp1. 0.9 kb product could be amplified from positive sample using primers 8 of hygromycin resistant gene 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27

Page 2: Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 · 1 1 Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 4 5 Fig. S1 Phenotypes of wild type and wp1

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Fig. S2 28

29

Fig S2 Subcellular localization of ValRS2 30 A, Subcellular localization of OsValRS2 in Nicotiana protoplasts. Bar = 20 µm. 31 B, Subcellular localization of AtValRS2 in rice protoplasts. Bar = 10 µm. 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58

Page 3: Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 · 1 1 Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 4 5 Fig. S1 Phenotypes of wild type and wp1

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Fig S3 59

60 Fig. S3 Alignment of OsValRS1, OsValRS2（WP1）, AtValRS1, AtValRS2 61

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Fig. S4 62

63 Fig. S4 Expression profiles of WP1, OsValRS1 and OsValRS3 from 64 http://ricexpro.dna.affrc.go.jp/ 65 66 67 68 69

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Fig S5 70

71 Fig. S5 Expression levels of chlorophyll synthesis genes in wild type and wp1 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105

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Fig. S6 106

107

Fig. S6 Western-blot analysis of chloroplast proteins in wild type and wp1w glumes. 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137

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Fig. S7 138

139 Fig. S7 Expression of nuclear-encoded chloroplast ribosomal protein genes in seedlings 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163

Page 8: Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 · 1 1 Supporting Information Figs S1–S8, Tables S1–S4 2 3 Fig. S1 4 5 Fig. S1 Phenotypes of wild type and wp1

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Fig. S8 164

165

Fig S8 TEM observation of mitochondria of wild type and wp1s. Bars = 200 nm. 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192

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Fig. S9 193

194

Fig. S9 Analysis mitochondrial ribosomes and mRNA levels of some mitochondrial genes. 195

A, Relative expression levels of mitochondrial 26S and 18S rRNAs in wp1 and wild type. 196

B, Western-blot analyses of mitochondrial ribosomal proteins in wp1 and wild type. 197

C, Relative expression levels of some mitochondrial genes in wp1 and wild type. Error bars 198

indicate SD (n = 3). 199

200

201 202 203 204 205 206 207 208 209 210 211 212

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Table S1 Agronomic characteristics of WT and wp1w 213 214

plant height

(cm) thousand kernel

weight (g) flag leaf length

(cm) flag leaf width

(cm) tiller

number spike length

(cm)

WT 77.4 ± 3.4 23.58 ± 0.34 30.5 ± 2.6 1.26 ± 0.08 5.4 ± 0.9 20.9 ±1.1

wp1w 76.6 ± 3.2 23.96 ± 0.26 29.7 ± 2.6 1.31 ± 0.05 5.4 ± 1.1 19.9 ± 0.9

215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250

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Table S2 Primers used in mapping 251 252

primer name Forward sequence Reverse sequence

RM82 TGCTTCTTGTCAATTCGCC CGACTCGTGGAGGTACGG

Bs1 GAGTGAACCCTGCTCCAG GAACACCACCTTATTTCTATTT

Bs2 CTGTGGAAAAGGACGCATAAA ATTGAATAGTGGCGGTGGATT

Bs8 GCTACTGTCTGCCTCCTTCG GGTTCAAAATGAGACGCTTG

Bs12 GGGCATTCTGTGCATTTT TGTCATTGTTTTGCCTTCTTA

Bs15 GCTTGTTGCTGCTGCTCT ACAGAATTATGTAGGGACTGC

Bs17 AGGGCATGTATCTAAACAAAGG AGCTACTCGAAGCATCCACTAA

Jc-1 GAAGTGATGCCACTCCATG TACCAATCAGCAAAGTCCC

253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275

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Table S3 Primers used in Real-time PCR 276 277

Primer name Forward sequence Reverse sequence

ACTIN TGGTCGTACCACAGGTATTG CCACATCTGCTGGAATGTGCTG

HEMA GATGCAATCACTGCTGGAAAGCGT CCATCTTGCCAGCACCAATCAACA

HEML AGAACAAAGGGCAGATTGCTGCTG TGTTTCGTCAAGTCACGGAGAGCA

HEMB TGGCATTGTCAGGGAAGATGGAGT CCAAAGCAGCACGTATTGCTCCAA

HEMC TCATTCCGAGGGCTATTGGCTTCA ACACTCTAGTTGGCCAATGGTGGA

HEME AATGGAGGCTTGCTTGAGCGAATG TTGTTACCAAGGCGTCTCCTTCCA

URO-D AGGCTTCCACTGACAGGTGTTGAT AAAGAACGCCAGGGTCAACATTCC

HEMF ACTGACTGCACGATGGCAGTATGA AGAGATCGAGCCATTCCTTTGGGT

CHLD TAGCACAGCTGTCAGAGTGGGTTT TTGCCAGCCACCTCAAGTATCTCA

CHLH GCACGGGAACTTGGCGTTTCATTA ACATGTCCTGGAGCTGCTTCTCAT

CHLI AGGGATGCTGAACTCAGGGTGAAA AAGTAGGACTCACGGAACGCCTTT

CHLM GCTTCATCTCCACGCAGTTCTACT GCAATGACGAATCGAAGACGCACA

CRD TGGATCTAACATGACACGCACCCA ACTGTAACGGCATTCTTCTCCGGT

DVR TTCTTCGAGAGGGTGATCAGGGAA GAAACTGGCAATGGCAGCCAAGAA

POR TCGTCGGCCTCGTCTGAGTTTATT AGGCCTCTCTCACTGAAAGCTGAA

CHLG CCAGCCACTGATGAAAGCAGCAAT AGAGCGCTAATACACTCGCGAACA

CAO ACACCTTCATCTGGGCTTCAAGGA AGATGCGTCGAACATTGCTTGGTG

DL-2 TCAGAAACTACCACTGCCAGAATC AGTCCCCCCAAAAGAAATCG

PsaA1 GGGAGGTGGCGAGTTAGTAG AATGCGTGAATGTGATGGAC

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PsaA2 TTATCTTCAACGAGCGGT TATCTCCAGGTCCTATTGTT

PsbD1 CTGCTACTGCTGTTTTCT GATGTTATGCTCTGCCTG

Rpoα GCGTCTTTATTATGGTCG TGTTCCTTCTGTTTCTCC

Rpoβ GTGGGGAACTTGCTTTAGG GCTTGTTGTATCCGTCTGA

Rpoβ' CATAGATTAGGCATACAGGC AATAGCGGGAGATAGGAG

Rpoβ'' AAAAGAGGAGGCTCGTGC GATGTTGGCTAAGTGATTGA

NADH2 ATCACTGTAGGACTTGGGTT TTTCCAGAAGAAGATGCC

NADH4 TCCTTATTGCTTATGCTGTC CCGTATGCTCCCATCTTTA

ATPα TCAAAAAGGGCAAGATGT TTGTAATGTAGCAGGGGAAT

ATPβ TTATTGGACCCGTGCTGG TTGCTTACCGTCAGTGTCTCG

RCA CTCTTCGTGCCCGTGTTTAC TCGGAGTTAGCGTCACCAAG

RbcL CAACTGTTTGGACTGATG GTTACCCACAATGGAAGT

Rps1 GGAAACGCCTCACAATCT TCCCACCTTATTCTCAAACC

Rps2 TTCGATTCCCTTTATACGC TTCCCGAGATACTGGTGG

Rps3 CCATCTCGCTCTTTCCTT CAATTTGCCTATCCTACCT

Rpl2 TCGTCTATTGGCATTGTGG GGTGGATTCGAGGATCTTAC

Rpl17 CGTCTTTCATATCGAGCCA GAGCAATCCAACCCGTAG

Ndh4(mt) ACAGGAACCACCGATTTA ACCATAGGCACTTTGACG

ccmFn CTTCATTCTTGGACCTCG AAATCTTCCTCGTGTATCG

26S rRNA ACAACGACCAATCCTGAA AGGCAGGCTTATACCATTAC

18SrRNA AAGACCGAAACTCAAAGGA TGTCAAGGGCTGGTAAGG

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278 Table S4 Primers used in vector construction 279

280 281 282 283 284 285 286 287 288 289 290 291 292

Primer name Sequence Used for

1390-WP1 Forward TTCTGCACTAGGTACCTGCAGATGGCGCTCGCGGGCGC

Ubi-WP1 Reverse TCTTAGAATTCCCGGGGATCCCTAATCAAAATTGTTATTTA

pA7-WP1-GFP Forward CCGCTCGAGATGGCGCTCGCGGGCGC

Subcellular localization

Reverse GGACTAGTGCATCAAAATTGTTATTTA

pA7-WP11-100-GFP Forward CCGCTCGAGATGGCGCTCGCGGGCGC

Subcellular localization

Reverse GGACTAGTGCCAGCGATCCGGTAACATTC

1305-WP1-GFP Forward AAGTCCGGAGCTAGCTCTAGA ATGGCGCTCGCGGGCGC

Subcellular localization

Reverse GCCCTTGCTCACCATGGATCCATCAAAATTGTTATTTA

pA7-AtVaLRS2-GFP Forward AAGTCCGGAGCTAGCTCTAGAATGATTCTCAAAACGGCTT

Subcellular localization

Reverse GCCCTTGCTCACCATGGATCCCTGAGACACTAAAGAAGTA

pA7-OsValRS1-GFP Forward CCGGACGTCATGTCCAGCGTTACCCCTGCTGCC

Subcellular localization

Reverse TCCCCCGGGATTATTGCCAGTTTTTGCATCTAG


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