利用微衛星序列作為台灣落花生栽培種DNA分子標誌

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(1)‫؀‬᨜ልᄐઔߒ (J. Taiwan Agric. Res.) 56(3):176-188 (2007). ճҔ༾ፁࢃ‫ׇ‬ӈբࣁѠ᡼ပ޸ғਭ୻ᅿDNAϩη኱ᇞ1 ഋণϘ2 ླྀߎᑫ3 මܿੇ2 ᎄ಍ໜ2 ֆܴণ2,4 㛧尐 ຫୃոΕᄘ८ᘋΕམࣟ௧ΕᔤอၼΕ‫ୃࣔܦ‬Ζ2007Ζ‫شܓ‬პᓡਣ‫ݧ‬٨‫܂‬੡‫؀‬᨜ᆵक़‫்س‬ ഛጟ DNA ։՗ᑑ፾Ζ‫؀‬᨜ልᄐઔߒ 56:176-188Ζ ‫ء‬ઔߒ‫׌‬૞‫ؾ‬ऱ੡৬‫م‬ഏփᆵक़‫்س‬ഛጟ១໢ૹᓤ‫ݧ‬٨ (simple sequence repeat, SSR) DNA ਐెᇷறፖᦸࡳ SSR ഗ‫ڂ‬ஆ᧢ฆൣ‫ݮ‬Δԫ٥ᙇ‫ش‬ഏփ 12 ଡᆵक़‫்س‬ഛጟ֗ 97 ଡ SSR ֧՗ኙΔ‫ ڇ‬77 ଡ‫ױ‬ᛧ൓堚ཐ࿨࣠ऱ SSR ᑑ፾ᨠྒྷࠩ 102 ଡය൅Δࠡխ‫ ڶ‬26 ଡ SSR ഗ‫ڂ‬ஆ‫ڇ‬ᑌ঴ၴ‫ܧ‬෼‫ڍ‬ীࢤΔࠀ‫ ڶ׊‬13 ଡ঴ጟറԫࢤය൅‫ࠎױ‬঴ጟᦸࡳΖ‫׼‬؆Δ‫ڶ‬ 6 ଡ SSR ഗ‫ڂ‬ஆၞ۩ᇞ‫ݧ‬ᒔᎁኙೝഗ‫ڂ‬ীኪΔ࿨࣠᧩‫ڍڶق‬ጟᣊীऱ᧢ฆΔ‫ץ‬ਔ໢ுᎨ ‫ڍ‬ীࢤΕૹᓤ‫ݧ‬٨୚ᑇ‫֗א‬լܶૹᓤ‫ݧ‬٨ഗ‫ڂ‬ஆΔ᧩‫ق‬ઌለ࣍ࠡ‫ה‬։՗ᑑ፾ߓอΔSSR ᑑ፾‫ڇ‬ഏփᆵक़‫்س‬ഛጟ‫ܧ‬෼ለ೏‫ڍ‬ীࢤֺ෷Ζ. ᙯᔣෟΚᆵक़‫்س‬ഛጟΕ։՗ᑑ፾Ε១໢ૹᓤ‫ݧ‬٨Ζ. ⓜ 岏 ᆵक़‫( س‬Arachis hypogaea L.) ᥆࣍ߤઝ (Fabaceae) ཬढΔ‫ؾ‬ছᎁ੡ದᄭ࣍তભ੊Δ‫ڇ‬ੲ‫ܓ‬ፂ ࠅতຝࢨॳ௅‫קݪ‬ຝ੡Գ቏֏Ιᆵक़‫س‬᥆ (Arachis) ‫א‬ীኪΕ‫چ‬෻։ؒ‫֗א‬ᠧٌᘣ‫ࢤٽ‬ऱլ‫ٵ‬Δ։ ੡԰ଡᆏ (section)Δ‫ץ‬ਔฆᔆ؄଍᧯ (allotetraploid) ்ഛጟक़‫( س‬AABB) (Hung et al. 1996)Ζઔߒ ᧩‫ױق‬౨௫֗‫ڰ‬ཚມ‫س‬ጟ A. duranensis (A genome) ࡉ A. ipaensis (B genome) ᠧٌ৵Δᤉۖ۞࿇ࢤ ਩‫᧯ۥ‬଍‫ף‬ऱ࿨࣠Ιᆵक़‫س‬௅ᖕ‫׌‬ၗक़‫ݧ‬ऱ‫ྤڶ‬։੡ࠟࠅጟ hypogaea ࡉ fastigiataΔ٦௅ᖕীኪ֗ ‫س‬९௽ࢤ hypogaea ‫ױ‬։੡ hypogaea ࡉ hirsute ࠟ᧢ጟΙfastigiata ‫ױ‬։੡ fastigiataΕvulgarisΕ aequatoriana ‫ ֗א‬peruviana ؄᧢ጟ (Moretzsohn et al. 2004)Ζ்ഛጟक़‫س‬ऱཬ௓ীኪ֗ልᢌࢤण ‫᧩ڶ‬ထऱ᧢ฆ‫ڇژ‬Δ‫܀‬ਢΔDNA ᐋ్ऱ᧢ฆ‫ ڕ‬RAPD (random amplified polymorphic DNA) ΕAFLP (amplified fragment length polymorphism) ‫ ֗א‬RFLP (restriction fragment length polymorphism) ։࣫ ࿨࣠ࠀլࣔ᧩Δ٣ছഏփᆵक़‫س‬ઔߒՈ‫ٵ‬ᑌ൓ࠩᆵक़‫س‬ᙊႚࣴฆ৫‫܅‬ΔᏁ૞ᗴᙇለՕၦ֧՗Δթ౨ ᛧ൓ለ‫ڍ‬ऱ‫ڍ‬ীࢤΔ‫܂א‬੡ᘣᒴᣂএ֗ጟ଺ᦸࡳհ‫ܒ‬ឰ (Fan et al. 1999)Ζທ‫ ګ‬DNA ᧢ฆ‫܅‬ᆵऱ‫ױ‬ ౨଺‫ץڂ‬ਔΚ1.२ᄭԲ଍᧯ፖ்ഛጟၴऱഗ‫( ੌڂ‬gene flow) ᎽᡶΖ2.்ഛጟᄭ۞֟ᑇᠧٌଡ᧯਩ ‫᧯ۥ‬଍‫ף‬Ζ3.ߛጟ‫ޗ‬றૹᓤࠌ‫ش‬࿛໢ԫࢨᓤ‫( ైڂٽ‬Moretzsohn et al. 2004) Ζ‫ڼڂ‬৸‫הࠡە‬։՗ᑑ ፾‫شܓ‬ऱ‫ࢤ۩ױ‬Ζ ෼վ DNA ։՗ᑑ፾‫ݾ‬๬ऱ࿇୶༼ࠎᄅऱ‫܂‬ढ‫ߜޏ‬࿜ฃ֗ᛵᇞढጟփࡉढጟၴऱዝ֏ᣂএΔ௅ ᖕլ‫ٵ‬ऱ PCR ‫ݾ‬๬੡ഗ៕ऱֱऄΔ‫ ڕ‬RAPDΕAFLPΕSSR (simple sequence repeat) ‫ ֗א‬ISSR 1. 2. 3. 4.. ۩ਙೃልᄐࡡ୉ᄎልᄐᇢ᧭ࢬઔߒ໴‫ܫ‬ร 2293 ᇆΖ൷࠹ֲཚΚ2007 ‫ ڣ‬8 ִ 5 ֲΖ ‫ิݾسࢬء‬ᆤ‫ش‬Գ୉Ε೫ઔߒ୉֗‫ิݾس‬ઔߒ୉ଫิ९Ζ‫؀‬᨜ ‫؀‬խᗼ ᣆ୽ၢΖ ‫܂ࢬء‬ढิ೫ઔߒ୉Ζ‫؀‬᨜ ‫؀‬խᗼ ᣆ୽ၢΖ ຏಛ‫ृ܂‬Δሽ՗ၡٙΚ[email protected]ΙႚటᖲΚ(04)23302806Ζ.

(2) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 177. (Inter-SSR) ࿛։ֱ࣫ऄΔբᆖ‫פګ‬ᚨ‫ڇش‬ᆵक़‫س‬ᙊႚࣴฆ৫ေ۷ (Hopkins et al. 1999) Δࠡխ SSR ‫ݧ‬٨ਢ‫ ط‬1-6 ଡᨕഗૹᓤඈ٨ิ‫ګ‬Δሙ‫ڇ܉‬ഗ‫ิڂ‬խΔࠠ‫ڶ‬೏ऱ‫ڍ‬ীࢤֺࠏΕ٥᧩ࢤᑑ፾Ε࿨࣠‫ױ‬ ᔾΕ։࣫୲࣐࿛ᚌរΖKrishna ࿛Գ (2004) ‫ شܓ‬18 ଡᘛ٠ᑑࡳऱ SSR ֧՗։࣫ 48 ଡ Valencia ഗ ‫ڂ‬ীऱᆵक़‫்س‬ഛጟΔԫ٥ྒྷ൓ 120 ଡլ‫ٵ‬ഗ‫ڂ‬ஆΔᆖመ‫ᦞףآ‬ጩᑇؓ݁ऄ (UPGMA) հႃᆢ։ ࣫Δ౨ജ೴։੡ 3 ଡᙊႚઌ‫ۿ‬ႃᆢΙMoretzsohn ࿛Գ (2004) ᙇ‫ ش‬12 ଡ SSR ֧՗։࣫ 96 ଡᆵक़‫س‬ ঴ጟΰߓαፖ 36 ଡ२ᄭມ‫س‬ጟΔມ‫س‬ጟො።ฆᔆ؄଍᧯ A. monticola (AABB)ΕDD ഗ‫ ิڂ‬A. gladuliferaΕ13 ଡ AA ഗ‫ڶิڂ‬ᣂऱ 2 ଍᧯ (2n=20) Ε2 ଡ 2n=18 ‫ ֗א‬10 ଡ BB ഗ‫ิڂ‬ऱມ‫س‬ጟΔ ৬‫م‬ᆵक़‫س‬঴ጟΰߓαᆢႃΔ‫܂ױ‬੡ߛጟՂᘣ‫ء‬ᙇᖗऱ೶‫ە‬Ζ‫ڼ‬؆ΔSSR ഗ‫ڂ‬ஆ Ah-041 ࠡխ 292 bp ኙೝഗ‫ڂ‬ী੡ AA ഗ‫ิڂ‬঴ጟΰߓαࢬ௽‫ڶ‬Δ᧩‫ ق‬SSR ᔞ‫ٽ‬ᆵक़‫س‬௿ాᙊႚ᧢ฆऱઔߒΖ ႚอ௅ᖕ؆।ীኪऱᦸࡳΔ౛ழ‫࠹࣐׊‬ᛩቼ‫ైڂ‬ᐙ᥼Δ‫ޓ‬ᏺ‫ܑܒף‬ՂհܺᣄΔ૿ኙኔᎾੌຏข ঴‫ྤޓ‬ऄሒࠩᦸࡳ‫ؾ‬ऱΔᏁ૞ࠌ‫ݾהࠡش‬๬‫ࣚ܌‬ംᠲΔDNA ਐె։࣫ਢਐࠠ‫ڶ‬ଡ᧯റԫࢤऱᙊႚ ิ‫ګ‬։࣫ (Jeffrey et al. 1985) Δլ࠹ࠩิ៣ຝ‫ۯ‬Ε࿇ߛழཚፖᛩቼ‫ైڂ‬ᐙ᥼ΔLevy ࿛Գ (1988) ଈ ‫ אڻ‬M13 ᕸပ᧯ಮ‫܃‬ᓡਣ᧯ (minisatellite) ‫ݧ‬٨Δ‫شܓ‬তֱᠧ‫ٽ‬ऄ৬ዌ཯क़ΕՕຽΕ႓ߤΕ਴ᖪ‫א‬ ֗ਸᖪ࿛ཬढढጟᗑ௽ DNA ਐెΔࠀ‫ױ‬ፖࠡ‫ה‬೯ढࡉპ‫س‬ढ‫ࢬڶ‬೴ሶΔ෼‫ ڇ‬DNA ਐె։࣫‫ش‬ຜ ‫ױ‬։੡ढጟዝ֏ࡉᙊႚቹᢜ৬‫ࠟم‬Օֱ૿Δ‫ץ‬ਔ঴ጟᦸࡳࡉഗ‫᧯ڂ‬։࣫࿛ኔᎾᚨ‫( ش‬Staub et al. 1999) Ζ‫ء‬ઔߒಾኙ‫ࢬء‬ഛߛᆵक़‫س‬঴ጟ‫؀‬ል 6 ࡉ 7 ᇆ֗ࠡઌᣂᘣᄭ٥ 12 ଡ঴ጟΰߓαΔࠌ‫ ش‬97 ଡ SSR ֧՗ิ‫ٽ‬ၞ۩։࣫Δࠡխ 77 ิᛧ൓‫ױ‬։࣫ය൅Δ26 ิࠠ‫ڍڶ‬ীࢤׂ੄Δ‫܂ױ‬੡঴ጟᦸࡳ ֗ߛጟ‫ޗ‬றᙇᖗऱ೶‫ࠉە‬ᖕΔਢഏփଈ‫شܓڻ‬റԫࢤ SSR ᑑ፾৬‫م‬ᆵक़‫்س‬ഛጟ DNA ਐెᇷறΖ. 㧟㠨咖㡈㽤 ֻྏՄफ़‫ݡ‬჌ֽ໚! ؄଍᧯ᆵक़‫்س‬ഛጟ (2n = 40) ‫؀‬ል 6 ᇆࡉ‫؀‬ል 7 ᇆ֗ࠡઌᣂᘣᒴΰቹ 1α‫؀‬ል 5 ᇆΕ‫؀‬ত 10 ᇆΕ‫؀‬ত 11 ᇆΕত‫ ߓޏ‬141ΕCJ444ΕNcAcc17127ΕPI 240553ΕPI 314817Εદक़‫۫֗א‬ఄ‫ػ׃‬٥ 12 ଡ঴ጟΰߓαΰ। 1α Δ94 ࡉ 95 ‫ڣ‬ཚ‫܂‬Δਊԫ౳ය்ٙጟ࣍ልᄐᇢ᧭ࢬᄵ৛ࡉ‫ض‬ၴΖ ८ᅕප‫פ‬ᄃ̶‫!ژ‬ ᆺׂ DNA ဇ࠷ֱऄ௅ᖕ Bornet ࡉ Branchard (2001) ऱ CTAB (cetyltrimethylammonium bromide) ֱऄၞ۩ΖSSR ։࣫Κ᜔࣍᧯ᗨ 20 ȝL ऱ֘ᚨයٙၞ۩Δ‫ ܶץ‬100 ng ഗ‫ ิڂ‬DNAΕ10 mM Tris–HCl (pH 8.3)Ε50 mM KClΕ2 mM MgCl2Ε200 ȝM dNTPsΕ0.6U Taq polymeraseΕ0.25 ȝM SSR primerΕ 5% DMSO ‫ ֗א‬0.5 ȝg/ȝL BSAΔ‫א‬ፋ‫ٽ‬䇹ຑ᠙֘ᚨᕴ (MJ Research PTC-200) ၞ۩֘ᚨΔ֘ᚨᄵ৫ ೶‫ ە‬Ferguson ࿛Գ (2004) ৬ᤜ࿓‫ݧ‬Δၲࡨ 94к˃3 ։ᤪ৵Δၞ۩ 40 ༛ᛩऱ 94к˃45 ઞΕ55к˃45 ઞΕ 72к˃1 ։ᤪΔ່৵ 72к˃10 ։᝻ΖࠉᖕছԳ࿇।հ SSR ֧՗‫ݧ‬٨Δਗᙇ֧՗យ‫ٽ‬ᄵ৫ 55-60кΔ PCR ឩᏺขढՕ՛ 500 bp ‫א‬ՀΔ‫׊‬ಾኙ 2 ۟ 4 ᨕഗ១໢ૹᓤ‫ݧ‬٨٥ 97 ิ SSR ֧՗ิ (He et al. 2003; Ferguson et al. 2004; Moretzsohn et al. 2005)Ζ ࿪‫̈́ڧ‬ᇆညࡔᐂ! 20 ȝL ፋ‫ٽ‬䇹ຑ᠙֘ᚨᏺཷऱ DNA ขढΔ‫ף‬Ե 4 ȝL 6X ऱሽࣺᒷᓢ෈ΰ0.1% xylene cyanol ࡉ 30% glycerolα Δ‫ א‬3% Agarose SFRTM ೏ᇞ࣫৫ᡯᓄ (Amresco Co., USA) փܶ 0.5 ȝg/mL ᄽ֏Ԭᙍ (EtBr) ਩‫ۥ‬Δ‫ ڇ‬1X TBE ᒷᓢ෈ (89 mM Tris-borate, pH 8.0 ࡉ 2 mM EDTA) խၞ۩ሽࣺΔሽࣺය ٙ੡ 150 V ၞ۩ 90 ։ᤪΔ‫א‬࿫؆ᗉ‫ܧ‬ቝ৵ધᙕሽࣺቹᢜΖ.

(3) ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. 178. ቹ 1. ᆵक़‫؀س‬ል 6 ᇆࡉ 7 ᇆհᢜߓΖ Fig. 1. The pedigree of peanut Tainung No. 6 and No. 7.. । 1. ࠎᇢᆵक़‫( س‬Arachis. hypogaeae) հ঴ጟΰߓα‫ז‬ᇆΕ‫ټ‬ጠ‫ཬ֗א‬ढী Table 1. Code, cultivar name, plant botanical type and scientific name of subspecies and variety of peanuts used for SSR analysis No 1 2 3 4 5 6 7 8 9 10 11 12. Variety (line) Tainung No. 5ΰ‫؀‬ል 5 ᇆα Tainung No. 6ΰ‫؀‬ል 6 ᇆα Tainung No. 7ΰ‫؀‬ል 7 ᇆα Tainan No. 10ΰ‫؀‬ত 10 ᇆα Tainan No. 11ΰ‫؀‬ত 11 ᇆα Nan Kai-si 141ΰত‫ ߓޏ‬141α CJ444 NC Acc 17127 PI 240553 PI 314817 Honghua 1ΰદक़α Spanish whiteΰ۫ఄ‫ػ׃‬α. Plant botanical type Spanish Spanish Spanish Spanish Spanish Spanish Spanish Virginia bunch Spanish Valencia Spanish Spanish. Subspecies & variety Fastigiata valgaris Fastigiata valgaris Fastigiata valgaris Fastigiata valgaris Fastigiata valgaris Fastigiata valgaris Fastigiata valgaris Hypogaea hypogaea Fastigiata valgaris Fastigiata Fastigiata Fastigiata valgaris Fastigiata valgaris.

(4) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 179. TTS ૄЯळᏴത̈́‫ؠ‬Ԕ! SSR ഗ‫ڂ‬ஆᙇཷ‫ شܓ‬pGEM-T Easy Vector System (Promega Co., USA) ௅ᖕᐗ೸৬ᤜ‫ޡ‬ᨏΔ࠷ 3 ȝL PCR ขढ‫ף‬Ե 5 ȝL 2X rapid ligation bufferΕ3U T4 DNA ligase ‫ ֗א‬50 ng pGEM-T Easy VectorΔ ෗ࡉ݁֌‫ڇ‬৛ᄵՀ֘ᚨ 1 ՛ழΔ‫ٵ‬ழ࠷ԫጥ ECOSTM ໏ٚาઽΰ墿‫ݾسس‬Δ‫؀‬᨜α Δ௅ᖕᐗ೸৬ᤜ ‫ޡ‬ᨏၞ۩᠏ী‫ش܂‬Δ໏ٚาઽಯ٧৵‫ף‬Ե֘ᚨ‫ګݙ‬ऱ൷‫ٽ‬ᔆ᧯ᔼᛯ 1 ઞΔ್Ղ࣋Ե٧Ղ 2 ։ᤪΔ‫ܛ‬ ‫֌݁ױ‬Ⴡ‫ ܉‬LB / Amp (LB broth with 1.5% agar and 50 ppm ampicillin) ؓࣨഛ塄ഗΔ࣍ 37кਁᄵᒣ ଙᆜഛ塄 12 ՛ழ‫א‬ՂΔሶֲਗᙇ‫ۥػ‬ပᆵΔࡡಜ‫؀‬᨜ंՓ௽‫ݾس‬ֆ‫׹‬ၞ۩ᇞ‫ݧ‬Δࢬ൓‫ݧ‬٨Ղႚ۟ SDSC Biology Workbench (http://workbench.sdsc.edu/) ጻీၞ۩ઌᣂֺኙ։࣫Ζ. 俟㨫 ར‫܅‬Ϡ TTS ᇾᄫ̶‫!ژ‬ ‫א‬ฤ‫֧ٽ‬՗९৫ 18~26 ᨕഗΕយ‫ٽ‬ᄵ৫ 55-60к‫ ֗א‬PCR ឩᏺขढՕ՛ 500 bp ‫א‬Հයٙհ 97 ิᆵक़‫ س‬SSR ֧՗Δኙ 12 ଡᇢ᧭‫ޗ‬றၞ۩։࣫ΰ। 1αΔ‫ޢ‬ଡધᙕය൅ᆖመ۟֟ 3 ‫ૹڻ‬ᓤऱ SSR ։࣫Δᗴᙇ‫ױ‬ᛧ൓堚ཐය൅Εය൅ᑇ‫֗ؾ‬Օ՛ࠠ‫ڶ‬᡹ࡳ٦෼ࢤऱ 77 ิ֧՗Δࠡխ‫ץ‬ਔ 28 ิಾኙ di-nucleotide ૹᓤ‫ݧ‬٨ΰga ૹᓤ‫ ڶ‬17 ิα Ε38 ิಾኙ tri-nucleotide ૹᓤ‫ݧ‬٨ΰtaa ૹᓤ‫ ڶ‬23 ิα Ε1 ิ tetra-nucleotideΰctca ૹᓤα‫ ֗א‬9 යܶ‫ ڶ‬2 ጟ repeat motif ऱૹᓤ‫ݧ‬٨Δႛ SSR ֧՗ PM-73 ‫آ‬ ࿆ሉૹᓤ‫ݧ‬٨ᣊীΰ। 2α ΰHe et al. 2003α Ζ‫ޢ‬ଡ SSR ֧՗ิ‫ױ‬ᛧ൓ 1~3 ଡ‫ױ‬ᙃᢝය൅ΰቹ 2α Δ‫א‬ SSR ֧՗ิ pPGSseq11E11, pPGPseq2G3, PM238, Ah-075, Ah-594 ྒྷ൓່‫ ڍ‬3 ଡය൅Δؓ݁ଖ੡ 1.33 ය / S S R ֧՗ ิΰ 1 යΚ56 ิΔ2 යΚ15 ิΔ3 යΚ5 ิ αΔԫ٥ྒྷ ൓ 102 ଡය ൅Δ SSR ᏺ ༏ ขढΰ ය൅αՕ՛ঞտ࣍ 100 ۟ 445 bp հၴΰ। 2αΖ ‫ޢ‬ଡ SSR ֧՗ิೠྒྷࠩ 1~6 ଡኙೝഗ‫ڂ‬Δࠡխ‫א‬ഗ‫ڂ‬ஆ pPGPseq5D5 ࡉ PM36 ᨠྒྷ່ࠩ‫ڍ‬ 6 ଡΰቹ 3αΔૠ‫ ڶ‬26 ิࠠ‫ڍڶ‬ীࢤׂ੄Δؓ݁ 1.7 ኙೝഗ‫ ڂ‬/ S SR ഗ‫ڂ‬ஆΖ‫ ڇ‬102 ଡᨠྒྷය൅᜔ ٥‫ ڶ‬33 ଡ‫ڍ‬ীࢤය൅પ‫᜔۾‬ᑇ 32%Ζ‫ڼ‬؆Δ‫ ڶ‬13 ଡ SSR ֧՗ิྒྷ൓঴ጟറԫࢤය൅ΰ। 3α Δ் ഛ঴ጟ PI 314817 ࡉ NCAcc17127 ։ܑᨠྒྷ່‫ ڍ‬5 ଡ঴ጟറԫࢤය൅Δࠡխ SSR ֧՗ิ pPGSseq9B4 ࡉ pPGSseq11G3 ‫ٵױ‬ழ೴ܑࠟृፖࠡ‫ה‬ᇢ᧭‫ޗ‬றΰቹ 4α Δࠡ‫ڻ‬੡ CJ444 ᨠྒྷ 4 ଡΔഏփߛ‫ګ‬঴ጟ ‫؀‬ত 10 ᇆႛ࿇෼ԫଡറԫࢤය൅ΰቹ 4α Δ‫܂ױ‬੡঴ጟᦸࡳ೶‫ە‬ΔPI314817 ࡉ NCAcc17127 ᝫ‫ٵ‬ழ ٥ ‫ ڶ‬4 ଡ ፖ ࠡ ‫ ה‬঴ ጟ ΰ ߓ α լ ‫ ٵ‬հ ௽ ௘ ය ൅ (pPGSseq9A7, pPGSseq19E9, pPGSseq16F1, pPGSseq10D4)Ζ ར‫܅‬Ϡ TTS ԔЕ̶‫!ژ‬ ‫ءڇ‬ኔ᧭ऱ 77 ิ SSR ֧՗։࣫Δ࿇෼ຝٝය൅ፖ He ࿛Գ (2003) ࡉ Moretzsohn ࿛Գ (2005) ऱ࿨࣠‫஁ڶ‬ฆΔ‫ץ‬ਔ‫ ڶ‬20 ิ֧՗ข‫س‬ऱය൅ᑇ‫ؾ‬ፖՕ՛Ղ‫஁ڶ‬ฆΔ‫׼‬؆Δ‫ ڶ‬6 ิ SSR ֧՗ኙࠀ‫آ‬ ಖሉቃཚׂ੄Օ՛ΰPM65, PM137, PM188, PM201, PM204, PM238α Δ࣍ਢಾኙຍࠟຝ։‫஁ڶ‬ฆऱ SSR ഗ‫ڂ‬ஆΔ։ܑਗᙇ 3 ଡ SSR ֧՗ิ֗ຝ։ PCR ขढ‫஁ڶ‬ฆऱᑌ঴ၞ۩ᇞ‫ݧ‬Δ‫شܓ‬೸ᄐ֏ pGEM-T Easy ཷ᧯ऴ൷ᙇཷ PCR ขढࠀ‫܂‬ᇞ‫ݧ‬ΰ। 4αΔ࿨࣠ᆵक़‫ س‬SSR ֧՗ิ pPGSseq17G6 ᨠ ྒྷࠩ 2 ิઌฆය൅Δ։ܑ੡ pPGSseq17G6a 275~296 bp ࡉ pPGSseq17G6b 236~241 bpΔ‫ڼ‬؆Δᒔᎁ ࠟጤ‫ݧ‬٨ፖᙇ‫ش‬ऱറԫࢤ SSR ֧՗‫ݧ‬٨ઌฤΔࠀ‫ݧڇ׊‬٨խ޲‫ڶ‬࿇෼ٚ۶ࣔ᧩ऱ TTG motifΙ.

(5) ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. 180. । 2. ᆵक़‫ س‬SSR ։՗ᑑ፾։࣫հ֧՗֗ PCR ขढ Table 2. List of SSR primers used in 12 peanut cultivars, and the products of PCR amplification. Primer. Size (bp). Number of bands. No. of distinguishable patterns. Primer. Size (bp). Number of bands. No. of distinguishable patterns. SSR Motif. pPGSseq11E11. 186. 3. 2. pPGSseq13E6A. 252. 1. 1. ga. pPGSseq14F4. 163. 2. 1. taa. taa. pPGSseq15C12. 274. 2. 3. pPGSseq14E10. 151. 1. 1. taa. ttg. pPGSseq16F1. 271. 1. 3. taa. pPGSseq15B1. 116. 1. pPGSseq17G6. 249. 2. 1. taa. pPGSseq16G8. 194. 1. 2. taa. 5. ttg. pPGPseq2C11. 264. 1. 1. taa/cac. SSR Motif. pPGSseq18B8. 254. 2. 1. taa. pPGPseq3A1. 238. 1. 2. taa. pPGSseq18G10. 254. 1. 1. ttg. pPGPseq2G3. 215. 3. 1. taa. pPGSseq9A7. 280. 1. 3. taa. pPGSseq11G3. 224. 1. 3. ctt. pPGPseq3F1. 290. 1. 1. ctca. pPGSseq16F10. 298. 1. 1. ga. pPGPseq3E10. 276. 1. 1. taa. pPGSseq15D3. 289. 1. 2. ga. pPGPseq7H6. 300. 1. 1. ctt. pPGPseq5D5. 274. 2. 6. ga. pPGPseq3D9. 292. 1. 1. ga/gt. pPGSseq18C2. 139. 1. 1. at. pPGSseq9F1. 240. 1. 1. at. pPGPseq3A8. 152. 1. 1. taa. pPGSseq11H1. 162. 1. 1. ga. pPGPseq2G4. 289. 1. 2. taa. pPGSseq12E10. 288. 1. 1. taa. pPGSseq19D9. 271. 1. 3. taa taa. pPGSseq17E3. 193. 1. 1. ctt. pPGPseq2D12B. 265. 1. 3. pPGSseq18A5. 268. 1. 1. at/taa. pPGSseq15E12. 135. 1. 1. at. pPGSseq19E9. 255. 1. 2. taa. pPGSseq15E8. 298. 1. 1. taa. pPGSseq9B4. 254. 1. 3. ttg/taa. pPGSseq12B6. 238. 1. 1. cta. pPGPseq1B9. 282. 2. 1. ga. pPGSseq15F12. 281. 1. 2. ga. pPGSseq12F7. 290. 1. 1. taa. pPGSseq18G9. 225. 2. 1. taa. pPGSseq19F4. 283. 1. 1. ga. PM-53. 116. 1. 1. at. pPGPseq8E12. 198. 1. 3. ttg/taa. pPGPseq8D9. 132. 2. 3. ctt. pPGPseq2F5. 262. 1. 1. taa. PM-73. 100. 1. 1. pPGSseq18G1. 277. 2. 1. taa. PM-210. 179. 1. 3. ct. pPGPseq2E6. 250. 2. 4. ga. PM65. 210. 1. 1. ct. PM3. 200. 1. 1. ga. PM137. 150. 1. 1. ga. PM36. 190. 2. 6. ga. PM188. 100. 1. 1. ga. PM45. 100. 1. 1. ga. PM201. 223. 2. 1. ct. PM145. 170. 1. 1. ct/ca. PM204. 214. 1. 3. ga. PM183. 100. 1. 5. ct. PM238. 170. 3. 1. ct. PM200. 150. 1. 1. ct/ca. Ah-075. 149. 3. 1. aca. Ah-041. 266. 1. 1. ctc/aac. Ah-325. 230. 1. 1. ttg. Ah-193. 445. 1. 1. aac/ga. Ah-594. 184. 3. 1. aac. Ah-558. 233. 2. 1. ttg. Ah-638. 268. 1. 1. aac. Ah 4-26. 150. 2. 3. ct. Ah-649. 220. 2. 1. aac. PM-15. 177. 1. 1. ga. Ah-715. 112. 1. 1. aac. PM-32. 103. 1. 1. ct.

(6) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 181. ቹ 2. ᆵक़‫( אس‬a). Ah-638Ε(b). Ah-649 ‫( ֗א‬c). Ah-594 ֧՗ឩᏺհ SSR ቹᢜΔ‫ޢ‬ଡ SSR ֧՗ิ։ܑྒྷ൓ 1-3 ଡය൅Ζΰ1~12 ੡ᆵक़‫س‬঴ጟ‫ז‬ᇆΔᇡߠ। 1α Fig. 2. Amplification profiles of 12 cultivars (lines) of peanut using (a). Ah-638, (b). Ah-649, and (c). Ah-594 as SSR primers revealed 1-3 fragments. (Line numbers at top refer to the number in Table1. M=100 bp DNA ladder)..

(7) 182. ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. ቹ 3. ‫( א‬a). PM36 ࡉ (b). pPGPseq5D5 ੡֧՗հ 12 ଡᆵक़‫ س‬SSR ‫ڍ‬ীࢤቹᢜΖΰ1~12 ੡ᆵक़‫س‬঴ጟ‫ז‬ᇆΔ ᇡߠ। 1α Fig. 3. The polymorphic SSR patterns of 12 cultivars (lines) of peanut obtained using primers PM36 and pPGPseq5D5. (Line numbers at top refer to the number in Table 1. M= 100 bp DNA ladder).. । 3. ᦸܑᆵक़‫س‬঴ጟΰߓαհ SSR ֧՗ Table 3. SSR primers for differentiating peanut cultivars Cultivar Tainan No.10 CJ444 NC Acc 17127 PI 314817. Primer revealing specific fragments pPGPseq2D12B pPGSseq18A5, pPGSseq13A10, pPGSseq16G8, pPGPseq5D5 pPGSseq11E11, pPGSseq9B4, pPGSseq11G3, PM36, Ah 4-26 pPGSseq9B4, pPGSseq11G3, pPGSseq15D3, pPGPseq8D9, PM-210.

(8) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 183. ቹ 4. ᦸܑᆵक़‫س‬঴ጟհ SSR ᑑ፾Δ‫( א‬a). pPGPseq2D12B ࡉ (b). pPGSseq11G3 ੡֧՗‫ױ‬։ܑᦸܑ‫؀‬ত 10 ᇆΕNCAcc17127 ‫ ֗א‬PI314817Ζΰ1~12 ੡ᆵक़‫س‬঴ጟ‫ז‬ᇆΔᇡߠ। 1α Fig. 4. SSR markers for identification of peanut cultivars Tainan No. 10, NCAcc17127, and PI314817 obtained using primers (a). pPGPseq2D12B and (b). pPGSseq11G3, respectively. (Line numbers at top refer to the number in Table 1. M= 100 bp DNA ladder). । 4. ഏփᆵक़‫்س‬ഛጟ SSR ֧՗‫ݧ‬٨Εׂ੄Օ՛Εૹᓤ‫ݧ‬٨ᣊী‫֗א‬ᇞ‫ݧ‬ᑌ঴ Table 4. Characterization of SSR loci in Taiwan cultivated peanut primer pairs, fragment size, repeat motif, and sequenced samples SSR locus pPGSseq17G6a pPGSseq17G6b pPGSseq9F1 PM-32 PM188 PM201a PM201b PM204. Primer pairs (5’ĺ3’) AACGACAACGACAACGACAA TCCACTATACAGTTGGGGGC GCCTAGTGTGCAAAGGTGCT CACCTGATGTGTAGTGAGGCA AGTGTTGGGTGTGAAAGTGG GGGACTGGGAACAGTGTTTATC GGGCTTCACTGCTTTTGATT TGCGACTTCTGAGAGGACAA CCTTTATAGAGGACCTTCCCTCTC GCCTATTTGGTATCGGCTCA. TGGGCCTAAACCCAACCTAT CCACAAACAGTGCAGCAATC z Variety numbers refer to the number in Table1.. Size (bp) 296/288/276/275. Repeat motif/ similar loci Ah-371. Varietyz 2, 3, 6, 7, 10. 241/236 240. Ah-371 (AT) 5. 2, 5, 10. 103. (CT)8(CT/G)(CT)7. 3, 8, 10. 100/102. (GA)7/6. 3, 4, 10. 223/225. (CT)19/20. 2, 5, 10. 138 224/220/218/214. PM201/PM347a (GA)24/22/21/19. 1, 3, 9, 10.

(9) 184. ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. pPGSseq9F1 ቃཚขढՕ՛ 296 bp ᥆࣍ AT ᠨᨕഗૹᓤΔ࿨࣠‫ڇ‬೶ᇢ঴ጟΰߓαᨠྒྷଖ੡ 240 bp ࠀ ࿇෼ (AT) 5 ऱૹᓤ‫ݧ‬٨ΙPM-32 ቃཚขढՕ՛੡ 90 bpΔᆖመᇞ‫ݧ‬৵ᒔᎁܶ‫( ڶ‬CT) 16 motif ࠀ‫׊‬ ‫ڇ‬լ‫ٵ‬঴ߓխ࿇෼ԫଡ໢ுᎨ‫ڍ‬ীࢤ (single nucleotide polymorphism, SNP)ΙPM188 ᆖᇞ‫ݧ‬৵൓व ขढՕ՛੡ 100/102 bpΔլ‫ٵ‬೶ᇢ‫ޗ‬றႛઌ஁ԫଡ GA ૹᓤ‫ݧ‬٨ΙPM201 ขढ᥆࣍ CT ᠨᨕഗૹᓤΔ Ոਢ࿇෼ࠟิය൅ข‫ࠡس‬ԫಖሉ੡ PM201aΔය൅Օ՛ਢ 223 ࡉ 225 bpΔլ‫ٵ‬ᑌ঴ၴઌ஁ԫଡ CT motifΔ‫׼‬ԫಖሉ੡ PM201b 138 bp ࠀ‫آ‬࿇෼‫᧩ࣔڶ‬ऱૹᓤ‫ݧ‬٨ΙPM204 ขढ᥆࣍ GA ᠨᨕഗૹ ᓤΔᇞ‫ݧ‬Օ՛੡ 214/218/220/224 ࿇෼‫ ڶ‬4 ଡኙೝഗ‫( ڂ‬GA) 24/22/21/19ΰቹ 5α Δኙೝഗ‫ڂ‬ᑇ‫ؾ‬೏ ࣍ऴ൷ᨠྒྷሽࣺቹᢜΖ. 岝嵥 ‫ء‬ኔ᧭ԫ٥ᗴᙇ‫ נ‬77 ิ SSR ֧՗ၞ۩։࣫ΰ। 2αΔࠡխ‫ א‬23 ิಾኙ taa ૹᓤ‫ݧ‬٨່‫ڍ‬Δࠡ ‫ڻ‬੡ 17 ิ ga ૹᓤ‫ݧ‬٨Δ‫ٵ‬ழ tri-nucleotide ૹᓤ‫ݧ‬٨ᑇ‫ؾ‬೏࣍ di-nucleotideΔ‫ ڇ‬Ferguson ࿛Գ(2004) ᆵक़‫ س‬SSR ‫ݧ‬٨։࣫խਐ‫נ‬Δࢬ‫ڶ‬։ᠦᦸࡳऱ 110 ଡᆵक़‫ س‬SSR ‫ݧ‬٨‫ א‬taa (29%) ፖ ga (28%) ૹ ᓤ‫ݧ‬٨ֺࠏ່೏Ι‫ڼ‬؆ΔMoretzsohn ࿛Գ (2005) ‫ شܓ‬EST ᇷற஄ေ۷ᆵक़‫س‬ഗ‫ิڂ‬խ tri-nucleotide ૹᓤ‫ݧ‬٨‫۾‬٤᧯ 73.2%Δdi-Εtera-‫ ֗א‬penta-nucleotide ૹᓤ‫ݧ‬٨։ܑ੡ 22.2%Ε3.3%‫ ֗א‬1.3%Δ ᧩‫ءق‬ኔ᧭֧՗ऱᙇᖗՕીฤ‫ٽ‬ᆵक़‫س‬ഗ‫ิڂ‬ऱ௽ࢤΖۖ‫ޢ‬ଡ SSR ֧՗ᨠྒྷࠩ 1-3 ଡය൅Δፖഏ ؆ᆵक़‫்س‬ഛጟؓ݁ 1~3.02 ઌ२ (Krishna et al. 2004)Δ‫܀‬ਢࡉ٣ছ႓࿛Գ (2001) ‫ֽشܓ‬ᒟࡉ‫ۏد‬ ऱ SSR ֧՗։࣫ᆵक़‫س‬ጟၴᠧٌጟΔ࿨࣠ᨠྒྷࠩ 4~11 ଡය൅‫᧩ࣔڶ‬ᆵ஁Δ‫ױ‬౨ऱ଺‫ڂ‬ਢ‫࣍ط‬ᆵक़ ‫்س‬ഛጟਢฆᔆ؄଍᧯Δ‫ֽאڼڂ‬ᒟ֗‫ ۏد‬SSR ֧՗ೠྒྷࠩᆵक़‫س‬լ‫ٵ‬ഗ‫ૹิڂ‬ᓤऱ SSR ഗ‫ڂ‬ ஆΖۖ‫ء‬ኔ᧭ආ‫ش‬ऱᆵक़‫ س‬SSR ֧՗‫ڍ‬ᑇਢ‫ૹشܓ‬ᓤ‫ݧ‬٨‫܂‬੡ᠧ‫ٽ‬൶ಾΔ۞ഗ‫ڂ‬஄խϘ຤࠷ϙࠠ ‫ૹڶ‬ᓤ‫ݧ‬٨ऱཷߓΔᆖመᇞ‫֧ࡉݧ‬՗๻ૠመ࿓Δࠀ‫׊‬ඈೈૹᓤऱ‫ݧ‬٨Δࢬ‫א‬റԫࢤለ೏ΰMoretzsohn et al. 2004αΖ. ቹ 5. ᆵक़‫ س‬SSR ഗ‫ڂ‬ஆ PM204 ‫ݧڍ‬٨ֺኙ։࣫Ζ Fig. 5. Multiple sequence alignment of peanut SSR locus PM204. The primer position is indicated by arrowhead, GA motif region by underline, and mismatch by dashed-line. TN5: Tainung No. 5, TN7: Tainung No.7..

(10) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 185. ‫ؾ‬ছբֆ‫܉‬၌መ 400 ଡक़‫ س‬SSR ഗ‫ڂ‬ஆΔ‫ޢ‬ଡ SSR ഗ‫ڂ‬ஆऱؓ݁ኙೝഗ‫ڂ‬ᑇ‫ؾ‬տ࣍ 4.25-6.9 (He et al. 2003, Krishna et al. 2004) Δ‫ء‬ኔ᧭ྒྷ൓ᑇᖕ 1.74 ኙೝഗ‫ ڂ‬/ SSR ഗ‫ڂ‬ஆᑇ‫݁ؓ࣍܅ؾ‬ଖΔ ‫ױ‬౨ऱᇞᤩਢ‫ߛڇ‬ጟመ࿓խ‫ڂ‬੡ૹ៿ࠌ‫֟ش‬ᑇऱᚌߜᘣ‫ء‬Δᖄીᆵक़‫்س‬ഛ঴ጟհᙊႚ᧢ฆ௿՛Δ ‫ ڕ‬SpanishWhite ‫ٵ‬ழਢ‫؀‬ል 4Ε5Ε6 ᇆऱᘣ‫( ء‬Huang et al. 1996)Δᖄીኙೝഗ‫ڂ‬ᑇ‫ؾ‬ऱ྇֟Δ‫ڇ‬ ՛ຽ்ഛ঴ጟᙊႚ᧢ฆऱઔߒխՈ࿇෼ઌ‫ۿ‬ऱൣ‫ݮ‬ΔԳ੡ऱߛጟᙇࢸທ‫ګ‬ऱᙊႚࣴฆऱ྇֟Δֺ֧ ጟᖄԵᄅऱഗ‫ڂ‬ীऱຒ৫ࠐऱ‫( ݶ‬Roussel et al. 2005) Δ‫ڼ‬؆Δ‫ء‬ኔ᧭ᙇ‫ ش‬3% Agarose SFRTM ᡯᓄ ‫܂‬੡ SSR ഗ‫ڂ‬ஆሽࣺ։࣫Δլᓵ‫ڇ‬ᓄ᧯ໂ፹ፖ਩‫ۥ‬Ղࠠ‫ڶ‬ᖙ‫܂‬១ঁऱᚌរ (Huang et al. 1996)Δ‫܀‬ ਢᇞ࣫৫ࡉᨋඕ৫ለ‫شܓ‬ႚอ 6%ፋ‫ׇ‬㋝₏ᓄ᧯஁Δທ‫ګ‬ᨠྒྷऱኙೝഗ‫ڂ‬ᑇ‫ؾ‬ೣ‫܅‬ΔSSR ഗ‫ڂ‬ஆᇞ ‫ݧ‬ᇷறՈᢞኔኙೝഗ‫ڂ‬ऱᑇ‫܅ࠩ࠹ؾ‬۷Ζ‫ڼ‬؆Δ‫ ڶ‬20 ิ֧՗ऱឩᏺขढྤऄ壄ᒔᦸܑΔ‫ܶץ‬ข‫س‬ լറԫऱ PCR ขढΔࢨਢ‫آ‬౨ᛧ൓堚ཐऱ࿨࣠Δ‫ױ‬౨ऱ଺‫֧ڶڂ‬՗๻ૠլߜΔທ‫ ګ‬PCR ֘ᚨ֧՗ យ‫ٽ‬ய෷լࠋΔຝٝऱྤயኙೝഗ‫( ڂ‬null allele) ੷۟‫ױ‬౨ਢ঴ጟΰߓα௽ᐛΔ‫܀‬սྤऄ‫ݙ‬٤ඈೈ ਢመ࿓խԳ੡ऱኔ᧭ᎄ஁Ζ ‫ء‬ኔ᧭խ‫ڍ‬ীࢤ֧՗ิࡉ‫ڍ‬ীࢤය൅ᑇ‫ؾ‬પ‫۾‬ᖞ᧯ 32%Δઌ२࣍ഏ؆்ഛጟक़‫ س‬SSR ઔߒऱ ࿨࣠ (34%) Δለૃ࿛Գ (1999) ‫ א‬RAPD ։࣫‫؀‬᨜ᆵक़‫س‬ጟ଺ऱ֧՗‫ڍ‬ীࢤֺ෷ 5.5%֗ය൅‫ڍ‬ী ࢤֺ෷ 9.9%೏‫ נ‬3 ଍‫א‬ՂΔ᧩‫ق‬ઌለ࣍ RAPD ‫֟྇אױ‬ኔ᧭ᑇ‫ۖؾ‬ሒࠩઌ‫ٵ‬ऱய࣠Δ‫ٵ‬ழ SSR ᑑ ፾լᓵ‫ױڇ‬ᔾࢤ٦෼ࢤՈࣔ᧩ᚌ࣍ RAPDΔ‫ڼڂ‬ᔞ‫܂ٽ‬੡ᆵक़‫س‬ᙊႚ᧢ฆհઔߒΖ்ഛጟᆵक़‫س‬։ ੡ 2 ࠅጟ Arachis hypogaeae spp. hypogaea ֗ Arachis hypogaeae spp. FastigiataΔࠉࠡཬढীኪ٥‫ױ‬ ։੡ SpanishΕValenciaΕVirginia bunchΕVirginia runner ؄ীΔૃ࿛Գ (1999) ‫ א‬11 ଡ RAPD ֧՗ ։࣫ 30 ଡഏփᆵक़‫்س‬ഛ঴ጟΰߓα Δឈྥ‫אױ‬೴։լ‫ٵ‬ऱߛጟߓอ֗‫آ‬೶ፖऱഏ؆֧ጟ‫ޗ‬றΔ‫܀‬ ਢ‫ٵ‬ԫߛጟߓอփլ‫ཬٵ‬ढী‫ޗ‬றথᆵ‫ڇ‬ઌ‫ٵ‬։ᆢΔ‫ء‬ኔ᧭ࢬࠌ‫ش‬ᆵक़‫؀س‬ል 6 ᇆࡉ 7 ᇆ֗ઌᣂᘣ ᒴ٥ 12 ଡ঴ጟΰߓα ΔೈԱ Virginia bunch ཬढী NCAcc17127 ࡉ Valencia ཬढী PI314817 ࠡ塒ຟ ਢ Spanish ཬढীΔࠀ‫ ڶٺृࠟ׊‬5 ଡ঴ጟറԫࢤය൅Δ֗ 4 ଡ٥ຏऱ‫ڍ‬ীࢤය൅Δ։ܑ‫אױ‬ፖ‫؀‬ ል 6 ᇆࡉ 7 ᇆ֗ࠡ‫ ה‬8 ଡ Spanish ཬढীऱߛጟߓอ೴ܑΔ᧩‫ق‬ઌለ࣍ RAPD ᑑ፾ΔSSR ᑑ፾‫ޓ‬ᔞ ‫்ٽ‬ഛጟᆵक़‫س‬ጟ଺ጥ෻հᚨ‫ش‬Ζ ‫ء‬ኔ᧭࿇෼ 26 ଡ SSR ഗ‫ڂ‬ஆऱ೶‫ە‬ᇷறլᏘ٤ࢨਢፖኔ᧭ᨠྒྷ࿨࣠‫᧩ࣔڶ‬ऱ‫נ‬Ե (He et al. 2003, Moretzsohn et al. 2005)Δࢬ‫א‬ਗᙇ 6 ଡᆵक़‫ س‬SSR ഗ‫ڂ‬ஆࠀಾኙຝ։ऱ೶ᇢ঴ጟΰߓαၞ۩ ᇞ‫ݧ‬։࣫Δ࿨࣠࿇෼ऱᒔፖᇷற‫ڶ‬լઌฤհ๠ΔpPGSseq17G6 ‫ٵ‬ழೠྒྷࠩ 2 ଡය൅Δࠡխ pPGSseq17G6a լ‫ٵ‬঴ጟΰߓαऱ஁ฆለՕΔ٣۩ᒔᎁ PCR ขढࠟጤऱ‫ݧ‬٨ፖ೶‫ە‬ᇷறԫીΔඈೈ ֧՗‫ݧ‬٨ᙑᎄऱ‫ױ‬౨ࢤΔ‫؀‬ል 6 ᇆ pPGSseq17G6a ‫ݧ‬٨ፖᇷற஄ᆵक़‫ س‬SSR ഗ‫ڂ‬ஆ Ah-371 ઌ‫ۿ‬Δ pPGSseq17G6b Ո‫ٵ‬ᑌ‫آ‬࿇෼ಖሉऱ TTG motifΔ᧩‫ق‬ຍԫ‫ݧ‬٨‫ֺڶ‬ለՕऱ᧢֏Δ‫ױ‬౨ਢ‫ڂ‬੡ᆵक़ ‫்س‬ഛጟਢฆᔆ؄଍᧯Δࢬ‫ א‬SSR ֧՗ᏺᘿഗ‫ิڂ‬խઌ‫܀ۿ‬լઌ‫ؾ࣍ٵ‬ᑑ SSR ‫ۯ‬ᆜΙpPGSseq9F1 ഗ‫ڂ‬ஆቃཚขढՕ՛੡ 296 bpΔۖഏփᆵक़‫்س‬ഛጟᨠྒྷࠩ 240 bp ׂ੄Δ‫ݧ‬٨խᒔኔ‫ ڶ‬SSR (AT)5 motifΔຍᑌऱ஁ฆᚨᇠਢ঴ጟ௽ࢤऱᣂএΔছԳઔߒխਐ‫ڶנ‬။‫ڍ‬ऱૹᓤ୚ᑇΔ‫ڍ‬ীࢤऱ࿓ ৫Ոለ೏Δۖᆵक़‫ س‬AT motif ‫່ڶ‬೏ऱؓ݁ 19.5 ଡૹᓤ୚ᑇ (Ferguson et al. 2004)ΙPM-32 ഗ‫ڂ‬ஆ ࠀ‫آ‬ಖሉ‫ݧ‬٨ૹᓤ motif ᣊীΔᆖመᙇཷᇞ‫ݧ‬৵ᒔࡳ᥆࣍ CT motifΔ‫ ڇ‬PI314817 ঴ߓ‫ݧ‬٨խ࿇෼໢ ԫுᎨ᧢ฆ (SNP) ਢ່՛ऱ‫ݧ‬٨஁ฆΔࠀྤऄ‫شܓ‬ཏຏሽࣺ։࣫࿇෼‫ڼ‬ᣊ‫ڍ‬ীࢤ᧢ฆΔႊ៶‫ط‬௽ ࡳऱுᎨփ֊䇹‫ ڕ‬Cel I ऱ‫ش܂‬Δթ౨‫ڇ‬ႚอሽࣺ։࣫խ᧩‫஁נق‬ฆΙ֧՗ PM188 ឩᏺขढᇞ‫ݧ‬൓ वය൅Օ՛։ܑਢ 100 ࡉ 102 bpΔլ‫ٵ‬ኙೝഗ‫ڂ‬ႛઌ஁ 2 bpΔլ୲࣐‫شܓ‬ᡯᓄሽࣺ‫஁נބ‬ฆΙ֧՗ PM201 ‫ٵ‬ழྒྷࠩࠟଡ‫ڍ‬ীࢤය൅Δ։ܑ‫ א‬PM201a ࡉ PM201b ।‫ق‬Δࠡխ PM201b ऱ‫ݧ‬٨‫آ‬࿇෼.

(11) ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. 186. CT motifΔࠟጤ‫ݧ‬٨ઌ‫࣍ۿ‬ᆵक़‫ س‬SSR ഗ‫ڂ‬ஆ PM201 ࡉ PM347aΔ‫࣍ط‬პᓡਣ‫ݧ‬٨Օၦሙ‫ڇ܉‬ഗ ‫ิڂ‬խΔ‫ױ‬౨ਢዝ֏መ࿓խಯ֏ໜ؈ૹᓤ‫ݧ‬٨ΙPM204 ഗ‫ڂ‬ஆ؄ଡᇞ‫ݧ‬ऱᑌ঴൅‫ڶ‬լ‫ٵ‬ऱኙೝഗ ‫ڂ‬Δࠀ‫ڇآ‬೏ᇞ࣫ᡯᓄሽࣺխ࿇෼஁ฆΔឈྥᡯᓄሽࣺ‫אױ‬១֏։࣫መ࿓Δ‫ݶף‬։࣫ய෷Δ‫܀‬ਢ‫ڂ‬ ‫܅૾ڼ‬Աය൅ऱᇞ࣫৫Δᖞ᧯ۖߢ SSR ։࣫ᚨ௅ᖕլ‫ٵ‬ऱኔ᧭Ꮑ‫ޣ‬ᙇᖗ‫ٽ‬ᔞऱ PCR ขढ։ֱ࣫ ‫ڤ‬Ζ‫ء‬ኔ᧭࿇෼்ഛጟᆵक़‫ س‬SSR ᑑ፾խ‫ڶژ‬๺‫ڍ‬ᙊႚ᧢ฆΔ‫ץ‬ਔ౒؈Εૹᓤ‫ڻ‬ᑇ஁ฆ‫ ֗א‬SNPΔ ለ RPAD ֗ ISSR ᑑ፾‫ޓ‬ᔞ‫ٽ‬ᆵक़‫س‬ᙊႚ᧢ฆઔߒΔ㯍൓ၞԫ‫ޡ‬ၲ࿇‫شܓ‬Δ‫౒܀‬រਢ SSR ։࣫‫ءګ‬ ೏Δૻࠫᚨ‫ش‬ऱཏሙࢤΖ‫؀‬᨜ᆵक़‫்س‬ഛጟऱᙊႚ᧢ฆ௿՛Δ࣐֧࿇ᙊႚ౤இࢤऱംᠲΔਚ२ᒴມ ‫س‬ጟ֧ၞ‫࣍ܗڶ‬ឩՕ᧢ฆ֗क़‫س‬㠩ఐፖᆺཌఐ‫ࢤݼ‬ጟ଺ऱ‫( شܓ‬Hung et al. 1999)Δ‫ױ‬ਢ DNA ᐋ‫ڻ‬ ऱ᧢ฆઌኙ‫܅‬ᆵΔ੡Աሒࠩߛጟፖጟ଺ጥ෻‫ؾ‬ऱΔᏁ૞৬‫م‬ᄅऱᙊႚ։࣫ߓอΔ‫ء‬ኔ᧭ਢഏփଈ‫ڻ‬ ආ‫ش‬ᆵक़‫ س‬SSR ᑑ፾Δ৬‫م‬ᆵक़‫ૹس‬ᓤ‫ݧ‬٨։࣫ੌ࿓Δ‫ޡॣګݙ‬ऱ DNA ਐెᇷறΔࠀ᧩‫ ق‬SSR ᑑ፾‫܂ױ‬੡ጟ଺ጥ෻ፖ঴ጟᦸࡳΙࠀ‫׊‬ಾኙᆵक़‫ڍس‬ীࢤհૹᓤ‫ݧ‬٨ၞ۩ᦸࡳፖ༴૪Δ‫ݦ‬ඨലࠐ౨ ‫࣍شܓ‬ᆵक़‫س‬ᙊႚቹᢜ৬‫م‬Ε։՗ᑑ፾᎖‫ߛܗ‬ጟΕ‫ࢤݼ‬ഗ‫ڂ‬ᙇཷ‫֗א‬ढጟփዝ֏ᣂএ৬‫م‬࿛‫ش‬ຜΖ. ㆤ䞷㠖䘊 (Literature cited) Bornet, B., and M. Branchard. 2001. Nonanchored Inter Simple Sequence Repeat (ISSR) Markers: Reproducible and specific tools for genome fingerprinting. Plant Mol. Biol. Rep. 19:209-215. Fan, M. J., S. F. Lo, J. Y. Wang, T. R. Shu, W. L. Tsaus, K. H. Yang, and Y. S. Cheng. 1999. Studies on the genetic relationship of peanut germplasm in Taiwan. J. Agric. Res. China 48:67-85. (in Chinese with English abstract) Ferguson, M. E., M. D. Burow, S. R. Schulze, P. J. Bramel, A. H. Paterson, S. Kresovich, and S. Mitchell. 2004. Microsatellite identification and characterization in peanut (A. hypogaea L.). Theor. Appl. Genet. 108:1064-1070. He, G. R. Meng, M. Newman, G. Gao, R. N. Pittman, and C. S. Prakash. 2003. Microsatellites as DNA markers in cultivated peanut (Arachis hypogaea L.) BMC Plant Biol. 3:3. Hopkins, M. S., A. M. Casa, T. Wang, S. E. Mitchell, R. E. Dean, G. D. Kochert, and S. Kresovich. 1999. Discovery and characterization of polymorphic simple sequence repeats (SSRs) in peanut. Crop Sci. 39:1243-1247. Huang, H. J., W. L. Tsaus, K. H. Yang, and J. N. Tsai. 1996. Evaluation of agronomic characteristics for the introduced wild peanut. J. Agric. Res. China 45:15-25. (in Chinese with English abstract) Huang, H. J., W. L. Tsaus, S. F. Lin, J. S. Hsieh, and J. N. Tsai. 1999. Identification and characteristic analysis of interspecific hybrids of peanut. J. Agric. Res. China 48:40-51. (in Chinese with English abstract) Huang, H. J., S. F. Lin, and J. S. Hsieh. 2001. Identification of the inconsistence between genomic constitution and phenotypic variation in interspecific hybrids (4x Ø 2x) of peanut with DNA marker. 2001. J. Agric. Res. China 50:12-24. (in Chinese with English abstract) Jeffreys, A. J., V. Wilson, and S. L. Thien. 1985. Individual-specific “fingerprinting” of human DNA. Nature 316:76-79..

(12) ᆵक़‫்س‬ഛጟ TTS ᑑ፾! 187. Krishna, G. K., J. Zhang, M. Burow, R. N. Pittman, S. G. Delikostadinov, Y. Lu, and N. Puppala. 2004. Genetic diversity analysis in Valencia peanut (Arachis hypogaea L.) using microsatellilte markers. Cell. Mol. Biol. Lett. 9:685-697. Moretzsohn Mde C., M. S. Hopkins, S. E. Mitchell, S. Kresovich, J. F. Valls, and M. E. Ferreira. 2004. Genetic diversity of peanut (Arachis hypogaea L.) and its wild relatives based on the analysis of hypervariable regions of the genome. BMC Plant Biol. 14:4-11. Moretzsohn Mde C., L. Leoi, K. Proite, P. M. Guimaraes, S. C. M. Leal-Bertioli, Ma. A. Gimenes, W. S. Martins, J. F. M. Valls, D. Grattapaglia, and D. J. Bertioli. 2005. A Microsatellite-based, gene-rich linkage map for the AA genome of Arachis (Fabaceae). Theor. Appl. Genet. 111:1060-1071. Roussel, V., L. Leisova, F. Exbrayat, Z. Stehno, and F. Balfourier. 2005. SSR allelic diversity changes in 480 European bread wheat varieties released from 1840 to 2000. Theor. Appl. Genet. 111:162-170. Staub, J. E., and F. C. Serquen. 1996. Genetic markers, map construction, and their application in plant breeding. Hortic. Sci. 31:729-739..

(13) 188. ‫؀‬᨜ልᄐઔߒ! ร 56 ࠴! ร 3 ཚ. Microsatellites as DNA Markers in Taiwan Cultivated Peanut1 Jen-Ren Chen2, Kin-Hsing Yang3, Tung-Hai Tseng3, Toong-Long Jeng3 and Min-Tze Wu2,4 Abstract Chen, J. R., K. H. Yang, T. H. Tseng, T. L. Jeng, and M. T. Wu. 2007. Microsatellites as DNA markers in Taiwan cultivated peanut. J. Taiwan Agric. Res. 56:176-188. The objectives of this work were to establish simple sequence repeat (SSR) DNA fingerprinting database and characterized SSR polymorphism in local peanut cultivars (Arachis hypogaea L.). In this investigation, 12 peanut cultivars were analyzed by using 97 SSR primer pairs. Seventy-seven SSR markers showed clear patterns and a total of 102 bands were recorded. Twenty-six SSR loci revealed polymorphism 13 of them were specific for cultivar identification. The average number of alleles per locus was 1.74, and up to 6 alleles were found at one locus. Six SSR loci in tested cultivars were sequenced for allele identification. The results indicated that different type of variation in SSR loci, including single nucleotide polymorphism (SNP), tandem repeat numbers, and non-repeat sequence locus. The results revealed that SSR markers produced a higher level of DNA polymorphism than other DNA markers in cultivated peanuts. Key words: Cultivated peanut (Arachis hypogaea L.), Molecular marker, Simple sequence repeat (SSR).. 1. Contribution No. 2293 from Agricultural Research Institute, Council of Agriculture. Accepted:August 5, 2007. 2. Respectively, Research Assistant ,Researcher and Senior researcher and Head of Biotechnology Division, ARI, Wufeng, Taichung, Taiwan, ROC. 3. Researcher, Crop Division, ARI, Wufeng, Taichung, Taiwan, ROC. 4. Corresponding auther, e-mail:[email protected]; Fax:(04)23302806..

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