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muom円hH
同 ωm‑AHh
寸ω門‑hF回︑
A.TIR ‑4.4
1 10kb ー+
No. of strainsー+
11.0 5.4
‑11.0 ‑8.4 ‑2.8 +‑2.4
kb ↓↓
3.4
ー争仏コ αコ 1 Fig 9. Variation bandi時patternprobed with TIR (A) and TNP (B) in Japanese lowland strains. Eleven strains in Type 1 was hybridized to TNP (B) and showed RFLPs among them.
3 11 36 No. of strains
C.TNP B.TNP A.TIR
的ω向凶hvH
同ω角同hH
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一 +‑
Type 1 kb 11.0
5.4 ‑
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LM
L臥 11.0
5.4
一 11.0
守一7.4 ‑2.8
kb
bコ <D 2 Fig 10. Variation banding pattern probed with TIR (A) and TNP (B and C) in Japanese upland strains. Twenty‑seven strains in Type 2 and twenty‑two strains in Type1
were hybridized to TNP(B)
and showed RFLPs among them.7 13
N
o. of strains 12 15N
o. of strains 1 22 27 N o. of strains‑ 凶 口 明 記 ︼ 凶
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R政J1MILLYFI<FCSDKIPCVLSPDRIDLSSRPAS亨LT叡3阻.GSG1GRGDSSAPP M L
Ku.Ds MDL一 一 … … … … 一 一 … … … 一 一 一 … … … 一 一 一 一 … … ー え 級/1H畏LRLRRGAGPHPαISPTSSSPSQ畏HRPHPPPLPPPPPSi¥AATATP1主主R
T雪 量色::uJs‑‑ー‑‑‑‑‑‑‑‑‑‑‑‑‑‑‑‑‑‑ー・鴨司ーー・・・ー‑‑‑・‑‑ーーーーーー，司ーrp‑‑ーー R滋J1PHPPRPPSTATP工 岱RP控PPPPS拙TAAP1HRYLHPL狩 郎 総 主PRSAL Ku.Ds・・a・百ーーー‑‑・ーー・咽‑‑‑ーーーーーーーー，ーーー宇田ーーー劃句情物也‑‑‑一ー一一明ゆ叩 え続1)PALHSLLPSLNV拡 探 ' DS箆DDSTWLQ1VG置LEE窓路島lQLVVVQSPVDEPD
雪S"D .... 安.."'SP lIP Ki.お必芳一一一一……‑‑SFNSLDSNG一 一 一 一 … … …‑‑‑1PNSPDVDPA .RJIu1 S鴎SS狩TQGEQPQPEPHPPP工VD宵 到LQIIおじDEEGRVN1VDDDEL YV G ** * * 玄.... だDW.....* .. .. LD*E(立民抑制 .. E*Y*
Ku.Ds LGETGGS区;LQKIDG窓SQ緬 ・ ー ‑LDWDSIIVSDVLDDEG琵VQVPT玄NEIYF 疋 滋J1LLGIぶAEDEA且 閥AQAAAATE倍以蕊紋忍総議E普及NHD附 脚 質VEPPV諮 玄
LGL**DE孟A N**lI G * G * " " 0 .. ** P 0 Ku.Ds NLGLNKGD給 総 波 及E・5一 一 …GSG叩αIAQGSLDT治資E恐 弘 明PCQDY R級r1VAGERMMVNDAl政:PSLV部TVYP獄KV!'RLAVRQFAlNEEFELWVI<ATDR ..."主会場V" PS会 G"倉 P潟支K FR"A"RQ A1 定F琵L金 書T Kuよ'Is1PD豆 総VVYN悶 慣SMQPGCLFPN寵 怒 思 議MRQYAIお思想・ELG1EVTSτ 必 級r1KKYVGACKGASDC:PWIM事犠RQADE‑RTVMVTI<FT祝 双TCTSSG翠RKTTT
常YVG CKG" DCP嘗合*"R T""V" 綬TCTSSG豆:RKTTT Ki.丘DsT京YVGYCKGG‑DC:PWR1YARE玄KKGLPT1VVAVLDDVHTCTSSGRRRTTT 先 鋭J1PTSA腎V及SKlHH1LRTDSGMGPKELQKRLQEDQKCKINYDTVAKGRSLAM PT* WVA脅A "L "MG"KELQ LQ I"YDTV*KG" 且*
Ku.Ds PTO倒 V A n 汲 貯LL蕊友好g船 為 路LQQTLQ廿HNVT1GYDTV撚:GKE笈主L ぷ 級J11QLQGS背 窓 間n髄LYRWRAAVM部SPGSV1窓1DTIEV以来VYF'NRITC孟L L*GSWEE"i' *LY W* AV* P SV認ID 1 DGK YF*RITCA*
Ku.Ds REL YGSWEESi'QLLYS腎X混且VIAVMPDSVIE工DV1L玄DG笈YYFSRITCAF R滋r1SPC1T盟、LTGCRPYLSVNST紅 混 乱 獄 忽 恥ASA1AIDGN榔 締 結AFGIT
マC1"慣 GCRPYLSV STA工蕊G総GHLASA 叩G NWMYド E宮 古F KuIJJl GPC1Sl湾 攻DGCRPYLSVDS冨!U.MGRWNm弘ASATGV開 問 問YPVI岱 明 ? え級r1DAETTDNWTWFM1QUぷAIC衣VSPLA1CTDAC岡山主主将まますFP禍 続IRE 主主T DNiI' WFM QL K""G 禽"*L昆IC*DA*KGL AV 亨すきA *RE Ku.Ds Q主主T亨DN引WFMKQL庶VVGD:舵LL孟ICSDAQ波 乱 綬IAVNEVFPY為 総 統 必 説/1α首 班.TQNL工 技YGGSVFQEMYPVARSYRAQV路 韮CMDTIlil<ACTDVALW
ぽ HL叩 "K* GS MYl'*AR YR V E ..吋.. *A Ku.Ds侵 攻 総 必G抑Vね笹IAGS… 滋 液yp轟A京昆YRRDVFEF膏IVSKVR泌す‑}II(工AEY R厳/1LDm依L工寄YRS使1lAEIK∞Y円 際 協 案 窃 ・ 期WIRD1KALP1 CELADTC LD. F衣食*WYRSG室羽 "IKCDY"TNN*込E おNW**D K LP*C*LA*
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R級J1REM1MTL官 繍 肌R1C議 官TGTILP主将.HQIぷARTRGLGi立.SVVHAD1TTA REM M L* RRRR1G K* G IL安VL L*ARTRGLG鼠.S"V" 0 * A Kuι'Is REMTMELFf銀系RR1GI登a.HGIILPSVIAIlぷARτ'RGLGHLSIVKCDNYMA 認 後J1EVWDNSSSfIA部羽衣食料SCTCQEWQHTG貯CQHALAVIASQQ1RDV紅
言V D"*** "fIV* *C叱 鰐Qr釘GKPCQH LA"1* Q 豆DV * Ku.Ds茸 漁DS指C釘 路IVVNぬELKQCSCEEWQH双 双PCQHGLALI1AQDSRDVGM R滋r1ENY1NPYYSV1uFW設AYNSIIEPLPDKSQWPKVDLPFγLGAPLAKRNLGR EN" YYS F 孟Y" "EP" D" S WP VD.... "孟P"及会R"LGR Ki.拭Ds窓NFVDDYYSTERFI<IAYS喪主VEPIGDRSFWPSVDFASGVF五P1ARゑ.GLGR 況 脱/1YRKL豆工KGCL窓口GGSKPKωω‑‑・‑‑‑‑伽蜘即物申ーー‑DC‑‑‑ωωωω制‑‑‑
"RK RIK*CLE G" ..京 C
Ku.Ds QR沼IffiIKSCLEGGS孟RNKST裂宣お玄KTKI<RIぷRQYTC雪NCG翠LG在員QSSYK
豆島三1ー・・・・切ー‑‑‑‑‑ー・・司‑‑ーーーーー・・一蜘情信一ーーー白畑叩叩側蜘知情ー‑‑‑‑軸 Ku.Ds CPLNGTK間 後 熔RINTT悶WIPKELRTSSQNVPVQPDVAEEVTEQELED R箆'u1 ーーーーー命働也岨ーーーー一ー‑…m ω ω ω ‑ ‑需主お照D‑時制蜘鍋嶋崎ーー事ーー』ー‑‑‑蜘‑‑‑
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必zl)sKKRLRIS且QQKQY
Fig 13. Comparison of amino acids sequences of mudrA gene and
rmuA gene.
‑42ー
A
Bc Ml 1 2 3 4 Ml 5 6
7 8Ml 9 M2
1.0kちゅ 0.6kb蜘1.0kbゅ 0.6幼ゅ0.6k~蜘lAωj
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1可
i Fig 14. RT酬PCRof rmuA gene transcript. A: 1.0kちRT錨PCRfragments co汀espondto the transcripts spliced with the 1 st and the 2nd introns. PCR products乙overedfrom the first ATG to the middle of the 3rd exson. 0.6主b仕agmentlost the 2nd exon with two intron. Lanes 1 and 3 were panicles at later stage. In contrast， lanes 2 and 4 were you設gpanicles. 日:Lanes 5 to 7 were tissues from the plants grown under 250 C(day) and 150 C (night). Lane 8 were tissue from the plant grown under 250 C (day) /20o C (night). C: R下PCRproduct obtained from calli.
A.
2 5 / 1 8 IR36
l
480詰p叩
B.
C.
ALTS
A1 2
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3
3 7 / 3 7 2 5 / 1 2 ( 1 week)
A1IR36
A1IR36
4
56
7ALTS‑rl
actin
円915. Activation of the rmuA gene. A: Cycling stress of lower temperature giveηfor a week activated the transcriptio打 ofthe rmuA ge持e.RT闘PCFミwas done with ALTS and r1 primer shown in panel C. B: Control for RT糊PCRreactioれ
by actin primers.
‑44‑
A. kb
2.2…
1.5 ‑
B. kb
M 1
M 1
2 3 4
伺峰一 Normallyspliced products
4暗一 Altenativespliced products
2 3 4 5 6
ヨ …
Genomic1 (1339)司 Genomic2(1203)
...̲ cDNA(294)
Fig 16. Alternative transcription amplified by RT‑PCR.
Panel A: 623f and RSTOP primers were used for RT‑PCR.
Normally spliced product including the exon 1 to 3 were shown with an arrow. Alternative spliced product lost the exo丹2.Lane 1， 3:宍NAsamples were extracted from mature leaves from A 1 and IR36 grown in green house. Lanes 2 and 4: RNA samples from flag leaves grown in green house. lane 5: RNA sample from mature leaves grown with cycling stress (250C for 12 hours and 120C for 12 hours). Panel B: primers
designed to amplify a portion of尺31gene including introns. Lanes 1 to 5 were same as panel A.しane6: Genomic DNA of A1. Amplified fragment from Pgi1 and Pgi2 were shown with arrows (genomic 1 and genomic 2).
‑45‑
NTR
A R4Iu1‑A1 240 335 5' C一四7叩GAGGAGGGAGAAGCTGGGA‑GCGGCATCGGCAGAGGCGACACCAGC毛TGCCTCCACATCGGCTTCGTCTCCGGCGCCGGCCCCCGCCCCTGCCAC * * *本*事訓ド***場*本****市*****判事****本****本****本******事*本*******本******ホ*****司院本*刻ドヨド本車事*司何事*羽炉本事本*謝辞*本当事本本***本 目玉STC71789 5' CCCTCGAGGAGGGAGAAGCTGGGAAGC詰GCATCGGCAGA告GCGACACCAGC誕TGCCTCCACATCGGCTTCGTCTCCGGCGCCGNCCCCCNCCCCTCCNAC 1 10 20 30 40 50 60 70 80 90 100 336 435 TCCCCCACCTCGTCCTCCCCCAGTCAGCGCCGCCG叩CCCCATCC詰CCTCCGCCACCGCCCCCATCCGCCGCCACCCCCACCCCTATCCGCCGCCCCCATC ***本*****本権準本*場事本**本******本**準**ホホ*刻ドホ*ホ***本*****事*****刻院本本**ホ本*本****本*******事本調院本当ド察本本判ド本事**本*担炉導***** TCCCCCACCT誕GTCCTCCCCCAGTNAGCGCCGCCGGCCCCATCCGCCTCCGGCACCG告CCCCATCCGCCGCCACCCCCACCCCTATCCGCNGCCCCCATC 101 110 120 130 140 150 160 170 180 190 200 436 514 CACC詰CTGCCCCTCGGTCAGCTCCTCCCGCCCTCCACTCCCTCCTCCCCTCCCTCAATGTGTGGTTAGGGTTTGGCTCC3' ***本*事事**本***事事中*****************本*******場*事場権**本****本**地*本第********本*****判事本*** CACCGCTGCCCCTCGGTCAGCTCCTCCCGCCCTCCACTCCCTCCTCCCCTCCCTCAATGTGTGGTTAG詰GTTTGGCTCC3' 201 210 220 230 240 250 260 2701st exon
AFmi
A R4Iu1‑A1 515 551 5' ATGGATGACTCCACCTGGCTACAGATTGTTGGGAGG 3' 本当事場**本***ホ****本*ホ****本******本*****事* 自主STC71789 5' ATGGATGACTCCACCTGGCTACAGATTGTTGGGAGG 3' 280 290 300 310
3 rd exon
1355 1453 A R4Iu1‑A1 5'玲ACAAAGTTTACTAATCATCACACGTGTACATCTAGCGGTAGAAGGAAGACTACCACACCAACCA‑GCGCCTGGGTTGCTTCCAAGGCCA刊CATATCC ホ*本*********事******************本調ド***ホ******本******本****ホ*本本**ホ*刻ド*本*****市場*本**事事事本**本調ドネ*司ド*司院本*本本*** B EST C71789 5' TGACAAAGTTTACT~ATCATCAÇAÇGTGTACATÇIA詰CGGTAGAAGGAAGACTACCACAACAACCAAGCGGCTGGNTTGCTTCCAAGGNCATCCATATCC 320 330 340 350 360 370 380 390 400 410 1~4 140 TTGGGACA‑GATTCAGGTAトGGGCCCTAAAGAACTGCAAAAAAGGTT3' 本****当事事******本*******場****ネ***本*******事事事**** TTGGGACAAGATTCAGGTATTGGGCCNTAAAGAACTTCAAAAAAGGTT 3' 420 430 440 450 460 Fig 17. Sequence comparison betweenまMu‑A1and rice EST， C71789. Gap was indicated with dash. Same nucleotides were shown with asterisks. Differences were markerd. 池山脳凶幽幽ム描鋪幽幽幽幽ト的凶. m
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Fig 19̲ Genomic fragments carring eHon 2. 6enomic PCR
山asdone with F同2and r4 primers which can amplify a
評議rtof coding意equencesincluding eHon 2.
‑48
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