ࠪࡠࠗ࠿࠽࠭࠽ߩ↢ᱺㆊ⒟ߦ⇣Ᏹߩ޽ࠆᄌ⇣૕ߩᨴ⦡૕⸃ᨆ ቟Ⓧ⦟㓉

全文

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Science Journal of Kanagawa University17 : 77-80 (2006)

©Research Institute for Integrated Science, Kanagawa University

ع⍴ ႎع 2005

ᐕᐲ␹ᄹᎹᄢቇ✚วℂቇ⎇ⓥᚲ౒ห⎇ⓥഥᚑ⺰ᢥ

ࠪࡠࠗ࠿࠽࠭࠽ߩ↢ᱺㆊ⒟ߦ⇣Ᏹߩ޽ࠆᄌ⇣૕ߩᨴ⦡૕⸃ᨆ

቟Ⓧ⦟㓉

1,4

㈬੗㤗ᧂ

1

㤥᫪ ፏ

2

᧻᳗ᐘᄢ

3

Analysis of Chromosome Behavior of Arabidopsis Mutants Defective in Reproductive Processes

Yoshitaka Azumi

1,4

, Asami Sakai

1

, Takashi Kuromori

2

and Sachihiro Matsunaga

3

1 Department of Biological Sciences, Faculty of Science, Kanagawa University, Hiratsuka-City, Kanagawa 259-1293, Japan

2 Plant Functional Genomics Group, RIKEN Genomic Science Center, RIKEN, Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan

3 Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita-ku, Osaka, Osaka 565-0871, Japan

4 To whom correspondence should be addressed. E. mail: yoshitk@info.kanagawa-u.ac.jp

Abstract:

From Arabidopsis mutant collections, more than 30 meiotic mutants have been isolated. However, the molecular mechanism of plant meiosis is still largely obscure. For the purpose of further understanding, we searched for new Arabidopsis meiotic mutants.

As a results of our collaboration, we found several new mutants, which were defective in reproductive processes. Since our main interest was in meiosis, we selected one meiotic mutant among them, and analyzed its chromosome behavior during meiosis.

Keywords:

Arabidopsis thaliana, reproduction, meiosis, chromosome, DAPI (4’,6-diamino -2-phenylindole)

ᐨ⺰

↢‛ߪ৻ߟߩ୘૕ߢ᳗㆙ߦ↢߈ߡ޿ߊߎߣߪߢ߈ߥ

޿ޕᰴߩ਎ઍ߳ߣ๮ߩᒁ߈⛮߉ࠍⴕ߁ߚ߼ޔ↢ᱺߦ ࠃߞߡሶቊࠍ↢ߺ಴ߔޕ߶ߣࠎߤߩ㜞╬↢‛ߪ㓽ᕈ

㈩஧ሶߣ㓶ᕈ㈩஧ሶ߇ว૕㧔ฃ♖㧕ߔࠆߎߣߦࠃߞ ߡᰴ਎ઍ߇↢ߺ಴ߐࠇࠆ᦭ᕈ↢ᱺࠍⴕ߁ޕฃ♖ߩ㓙 ߦᨴ⦡૕ߩ୚ട߇⿠ߎࠆ߇ޔ਎ઍࠍㅢߓߡᨴ⦡૕ᢙ ࠍ৻ቯߦߔࠆߚ߼ߦޔᷫᢙಽⵚࠍⴕ޿ޔ੍߼㈩஧ሶ ߩᨴ⦡૕ᢙࠍඨᷫߐߖߡ߅ߊޕᷫᢙಽⵚߪߖߞ߆ߊ

ੑ⚵޽ࠆᨴ⦡૕ࠍ৻⚵ߦᷫࠄߔಽⵚߢޔਇนᰳߥ߽

ߩߢߪ޽ࠆ߇ޔ৻ᧄߢ߽ᨴ⦡૕߇ᄙ߆ߞߚࠅዋߥ ߆ߞߚࠅߒߡ߽⥌๮⊛ߥᓇ㗀߇⃻ࠇޔᄢ߈ߥෂ㒾ࠍ

઻߁ಽⵚߢ޽ࠆޕ

ᷫᢙಽⵚߪ৻ᐲߩᨴ⦡૕ߩⶄ⵾ߩᓟޔੑᐲߩಽⵚ ߇⿠ߎࠆޕߘࠇߙࠇ╙৻ಽⵚޔ╙ੑಽⵚߣ๭߫ࠇޔ ೨ᦼޔਛᦼޔᓟᦼޔ⚳ᦼ߆ࠄߥࠆޕᷫᢙ╙৻ಽⵚߩ ೨ᦼߪޔᨴ⦡૕߇⚦޿♻⁁ߦ⷗߃ᆎ߼ࠆ⚦♻ᦼ㧔࡟

ࡊ࠻࠹ࡦᦼ㧕ޔ⋧หᨴ⦡૕߇ࡍࠕ࡝ࡦࠣߒᆎ߼ࠆว♻

ᦼ㧔ࠩࠗࠧ࠹ࡦᦼ㧕ޔቢᚑߒߚࠪ࠽ࡊ࠻ࡀࡑ᭴ㅧࠍ੺

ߒߡ⋧หᨴ⦡૕ห჻߇੕޿ߦធ⌕ߔࠆᄥ♻ᦼ㧔ࡄࠠ

࠹ࡦᦼ㧕ޔࠪ࠽ࡊ࠻ࡀࡑ᭴ㅧ߇፣უߒᨴ⦡૕߇ߐࠄߦ

ಝ❗ߔࠆⶄ♻ᦼ㧔࠺ࠖࡊࡠ࠹ࡦᦼ㧕ޔಝ❗߇߶߷ቢᚑ ߔࠆ⒖േᦼ㧔࠺ࠖࠕࠠࡀࠪࠬᦼ㧕ߩ5ߟߩᤨᦼߦಽ ߌࠄࠇࠆޕ

ᷫᢙಽⵚߣ૕⚦⢩ಽⵚߩ㗼⪺ߥ㆑޿ߪ╙৻ಽⵚ೨ ᦼߦ⋧หᨴ⦡૕߇ኻวߔࠆߎߣߢޔߎߩߎߣ߇⋧ห ᨴ⦡૕ߩᱜᏱߥಽ㈩ߩ೨ឭ᧦ઙߣߥࠆޕ⋧หᨴ⦡૕

ߩኻว߇ቢᚑߔࠆ߹ߢߦߪޔ࠹ࡠࡔࠕߩࠢ࡜ࠬ࠲࡝

ࡦࠣޔ⋧หᨴ⦡૕ߩਗ೉㧔alignment, juxtaposition㧕ޔ

ᷫᢙಽⵚᦼ⋧ห⚵ߺ឵߃ޔࠪ࠽ࡊ࠻ࡀࡑ᭴ㅧߩቢᚑ ߣ޿ߞߚࡊࡠ࠮ࠬ߇޽ࠆޕߎߩߥ߆ߢ⋧ห⚵ߺ឵߃ ߦ㑐ߔࠆ⎇ⓥ߇ᦨ߽ㅴࠎߢ޿ࠆޕ㉂Უߥߤߩ⎇ⓥ߆ ࠄ⋧ห⚵ߺ឵߃෻ᔕߪ SPO11 Ⱞ⊕⾰ߦࠃࠆੑᧄ㎮

DNA ಾᢿ߆ࠄ㐿ᆎߔࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ 1)ޕߘ ߩᓟޔ৻ᧄ㎮DNA ㇱಽ߇೥ࠅ಴ߐࠇޔߘߩ৻ᧄ㎮

߇⋧หᨴ⦡૕ߩ⋧หㇱಽߣ⚵឵߃ࠍ⿠ߎߒޔ෻ᔕ߇ ㅴ߻ߣ޿ߞߚࡕ࠺࡞߇ឭ᩺ߐࠇߡ޿ࠆ2)ޕ

㉂Უ߿േ‛ߢߪ SPO11 ㆮવሶߪࠥࡁࡓᒰߚࠅ㧝 ߟߒ߆⷗ߟ߆ߞߡ޿ߥ޿߇ޔࠪࡠࠗ࠿࠽࠭࠽ߢߪ 3 ߟ ߩ ⋧ ห ㆮ વ ሶ ޔAtSPO11-1ޔAtSPO11-2ޔ AtSPO11-3߇ሽ࿷ߔࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ3)ޕ

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78 Science Journal of Kanagawa University Vol. 17, 2006

AtSPO11-1ߪታ㓙ߦ⋧ห⚵ߺ឵߃ߩ㐿ᆎߦᔅⷐߢޔ ߎߩㆮવሶߩᄌ⇣૕ߢߪ⋧หᨴ⦡૕ߩኻว߇⿠ߎࠄ ߥ޿ߎߣ߇⍮ࠄࠇߡ޿ࠆ4)ޕAtSPO11-3ߩᄌ⇣૕ߢ ߪᩕ㙃↢㐳ߦ⇣Ᏹ߇⷗ࠄࠇࠆߎߣ߆ࠄ૕⚦⢩ಽⵚᤨ

ߦᯏ⢻ߒߡ޿ࠆߣ⠨߃ࠄࠇߡ޿ࠆ5)ޕAtSPO11-3ߣ ߩ㘃ૃᕈ߆ࠄ AtSPO11-2 ߽૕⚦⢩ಽⵚߦ㑐ਈߔࠆ ߣ⠨߃ࠄࠇߡ޿ߚ߇ޔ߹ߛታ㓙ߦߪ⺞ߴࠄࠇߡ޿ߥ ߆ߞߚޕߘߎߢᚒޘߪ AtSPO11-2 ߩᄌ⇣૕ࠍ౉ᚻ ߒޔߘߩ⴫⃻ဳࠍ⸃ᨆߒߚߣߎࠈޔ⋧หᨴ⦡૕ߩኻ วߦᔅ㗇ߢ޽ࠆߎߣ߇್᣿ߒߚޕ

᧚ᢱߣᣇᴺ

ታ㛎ᬀ‛

ࠪࡠࠗ࠿࠽࠭࠽㧔Arabidopsis thaliana㧕ߩ㊁↢ဳ

෸߮ߘߩᄌ⇣૕ߪޔ␹ᄹᎹᄢቇ࡮ᐔႦࠠࡖࡦࡄࠬౝ

ߩᬀ‛⢒ᚑ᫟ౝߢᩱၭߒߚޕ60mmolm-2s-1ߩ⊕⦡శޔ 14ᤨ㑆10ᤨ㑆ߩ᣿ᥧ๟ᦼޔ᳇᷷24͠ޔḨᐲ60㧑 ߩ᧦ઙਅߢޔࡂࠗࡐࡀ࠶ࠢࠬߣ MS ၭ࿾ࠍ੤੕ߦޔ ㅳߦ৻ᐲਈ߃ߥ߇ࠄᩱၭߒߚޕ᠞⒳ᓟޔ5㧙7ㅳ㑆⋡

ߩᬀ‛ߩ⧎ᐨࠍណขߒޔࡈࠔ࡯ࡑ࡯ᶧ㧔Ethanol, Acetic Acid; 3:1㧕ਛޔቶ᷷ߢ20ᤨ㑆⒟ᐲ⟎ߊߎߣ ߦࠃߞߡ࿕ቯߒߚޕߘߩᓟߪ-20͠ߢ଻ሽߒߚޕ ᶖൻዷ㐿ᴺ

⧎☳Უ⚦⢩ߩᨴ⦡૕⹜ᢱ૞⵾ߪ Azumi ࠄߩᣇᴺ6) ߦᓥߞߚޕ࿕ቯߒߚ⹜ᢱࠍ10 mMࠢࠛࡦ㉄✭ⴣᶧ 㧔pH4.5㧕ਛߢᵞߞߚᓟޔcytohelicase㧔Sigma㧕ޔ cellulase “ONOZUKA” R-10㧔Yakult㧕ޔpectolyase 㧔Kikkoman㧕㧔ฦ0.4%㧔w/v㧕㧕ࠍ฽߻ห✭ⴣᶧਛޔ ㆡቱ⣕᳇ࠍߒߥ߇ࠄޔ37͠3 ᤨ㑆଻᷷ߒޔ⚦⢩ოࠍ ᶖൻߒߚޕห✭ⴣᶧߢᵞߞߚᓟޔ4͠ߢ଻ሽߒߚޕ ᶖൻߒߚ⧎ᐨࠍࠪࡖ࡯࡟਄ߩ 60㧑㈶㉄ਛߦ⒖ߒߚ

ᓟޔㆡᒰߥᄢ߈ߐߩ⭫ࠍขࠅ಴ߒޔหߓߊ60㧑㈶㉄

ࠍṢਅߒߚࠬ࡜ࠗ࠼ࠟ࡜ࠬ਄ߢ⸃೬ߒߚޕ⫎ࠍߟ߱

ߒߡ⧎☳Უ⚦⢩ࠍ᜛ᢔߐߖߚߩߜޔ45͠ߩࡎ࠶࠻ࡊ

࡟࡯࠻਄ߦ1ಽ㑆㕒⟎ߒߚޕ᳖಄ߒߚࡈࠔ࡯ࡑ࡯ᶧ ࠍ๟࿐ߦṢਅߒޔ✭߿߆ߦᷙ๺ߐߖߚᓟޔࡈࠔ࡯ࡑ࡯

ᶧࠍᝥߡޔࠬ࡜ࠗ࠼ࠍੇ῎ߐߖߚޕDAPI ṁᶧ

Fig. 1. ㊁↢ဳࠪࡠࠗ࠿࠽࠭࠽ߩ⧎☳Უ⚦⢩ߩᷫᢙಽ

ⵚᦼᨴ⦡૕௝. ㊁↢ဳߩࠪࡠࠗ࠿࠽࠭࠽ߩ⧎ᐨࠍណ ขߒ, ࡈࠔ࡯ࡑ࡯ᶧߢ࿕ቯߒߚᓟ, ㆡᒰߥᄢ߈ߐߩ⭫

ࠍㆬ߮, ᶖൻዷ㐿ᴺߦࠃߞߡᨴ⦡૕⹜ᢱࠍ૞⵾ߒߚ. ᨴ⦡૕ߪDAPIߢᨴ⦡ߒߚ. A; ࡟ࡊ࠻࠹ࡦᦼ, B; ࠩࠗ

ࠧ࠹ࡦᦼ, C; ࡄࠠ࠹ࡦᦼ, D; ࠺ࠖࡊࡠ࠹ࡦᦼ, E; ࠺ࠖ

ࠕࠠࡀࠪࠬᦼ, F; ╙৻ಽⵚਛᦼ, G; ╙৻ಽⵚᓟᦼ, H;

╙৻ಽⵚ⚳ᦼ, I; ╙ੑಽⵚ೨ᦼ, J; ╙ੑಽⵚਛᦼ, K;

╙ੑಽⵚᓟᦼ, L; ╙ੑಽⵚ⚳ᦼ.

Fig. 2. atspo11-2 ᄌ⇣૕ߩ⧎☳Უ⚦⢩ߩᷫᢙಽⵚᦼ ᨴ⦡૕௝. ㊁↢ဳߩ႐วߣห᭽ߦ, atspo11-2 ߩ⧎ᐨ ࠍណขߒ, ᶖൻዷ㐿ᴺߦࠃߞߡᨴ⦡૕⹜ᢱࠍ૞⵾ߒ ߚ. A-C; ࡟ࡊ࠻࠹ࡦᦼ߆ࠄࠩࠗࠧ࠹ࡦᦼ, D-E; ࠺ࠖ

ࡊࡠ࠹ࡦᦼ, F-G; ࠺ࠖࠕࠠࡀࠪࠬᦼ, H; ╙৻ಽⵚਛ ᦼ, I; ╙৻ಽⵚᓟᦼ, J-K; ╙৻ಽⵚ⚳ᦼ, L; ╙ੑಽⵚ ೨ᦼ.

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቟Ⓧ㧘㈬੗ ઁ: ࠪࡠࠗ࠿࠽࠭࠽ߩ↢ᱺㆊ⒟ᄌ⇣૕ߩᨴ⦡૕⸃ᨆ 79

㧔Vector㧕ࠍṢਅߒޔࠞࡃ࡯ࠟ࡜ࠬࠍタߖߚᓟޔ㗼ᓸ

㏜㧔ࠝ࡝ࡦࡄࠬ㧮㨄61㧕ߢⷰኤߒߚޕ

⚿ᨐ

ET3962 (atspo11-2-1)

SALK⎇ⓥᚲߩ࠻࡜ࡦࠬࡐ࠱ࡦ࠲ࠡࡦࠣ࡜ࠗࡦߩਛ ߦ AtSPO11-2 ㆮવሶߦ࠻࡜ࡦࠬࡐ࠱ࡦ߇ᝌ౉ߐࠇ ߡ޿ࠆᩣET3962ࠍ⷗޿಴ߒߚޕߘߎߢߎߩᩣࠍข ࠅነߖޔ⴫⃻ဳࠍ⸃ᨆߔࠆߎߣߦߒߚޕ

ᩕ㙃ᚑ㐳ᦼߦ߅޿ߡߪ㊁↢ဳߣหߓߊ⦟ߊ↢㐳ߒޔ

⪲ߩᒻ⁁࡮ᨎᢙޔ⨲ਂޔ↢㐳ㅦᐲߥߤߦ․ߦ⇣Ᏹߪ

⷗ߟ߆ࠄߥ߆ߞߚޕ↢ᱺᚑ㐳ᦼߦ߅޿ߡ߽ޔ⪬ޔ⧎

ᑯޔ㓶ߒߴޔ㓽ߒߴߩ⧎ེቭߩಽൻߢߪޔᒻ⁁߿ᢙ ߦ⇣Ᏹߪ⷗ࠄࠇߥ߆ߞߚޕߒ߆ߒޔߎߩᄌ⇣૕ߩ㕽 ߪ⊒㆐ߖߕޔ⒳ሶߪ߶ߣࠎߤᓧࠆߎߣ߇಴᧪ߥ߆ߞ ߚޕ↢ᱺ⚦⢩ߦ⇣Ᏹ߇޽ࠆߎߣ߇੍ᗐߐࠇߚߩߢޔ

⧎☳Უ⚦⢩ߦ߅ߌࠆᷫᢙಽⵚᤨߩᨴ⦡૕ࠍⷰኤߔࠆ ߎߣߦߒߚޕ

atspo11-2-1ߩᨴ⦡૕ⷰኤ

㊁↢ဳߩࠪࡠࠗ࠿࠽࠭࠽ߩᨴ⦡૕௝ߦ㑐ߒߡߪ Rossࠄߩ⹦ߒ޿ႎ๔߇޽ࠆ7)ޕ⚦⢩ოࠍᶖൻߒߚᓟޔ

ࠬ࡜ࠗ࠼ࠟ࡜ࠬ਄ߦᨴ⦡૕ࠍዷ㐿ߔࠆᶖൻዷ㐿ᴺߦ ࠃߞߡᨴ⦡૕⹜ᢱࠍ૞⵾ߒޔᚒޘ߽ห᭽ߩⷰኤ⚿ᨐ ߇ᓧࠄࠇߚޕ╙৻ಽⵚ೨ᦼߢߪޔ♻⁁ߦᨴ⦡૕߇ⷰ

ኤߐࠇᆎ߼ߚ࡟ࡊ࠻࠹ࡦᦼ㧔Fig.1A㧕ޔࡍࠕ࡝ࡦࠣ

߇⿠߈ߡ޿ࠆߣ੍ᗐߐࠇࠆࠩࠗࠧ࠹ࡦᦼ㧔Fig.1B㧕ޔ ኻว߇ቢᚑߒߚࡄࠠ࠹ࡦᦼ㧔Fig.1C㧕ޔࠪ࠽ࡊ࠻ࡀ ࡑ᭴ ㅧ߇፣ უߒ ߡࠁߊㆊ⒟ ߩ࠺ ࠖࡊ ࡠ࠹ࡦ ᦼ 㧔Fig.1D㧕ޔಝ❗߇߶߷ቢᚑߒߚ࠺ࠖࠕࠠࡀࠪࠬᦼ 㧔Fig.1E㧕ߥߤ߇ⷰኤߐࠇߚޕ

ߎߩᄌ⇣૕ߢ߽ޔᷫᢙಽⵚᦼߦ౉ࠅޔ࡟ࡊ࠻࠹ࡦ ᦼ ߩ ᨴ ⦡ ૕ ߪᢙ ᄙߊ ⷰ ኤ ߔ ࠆ ߎ ߣ ߇ ߢ ߈ ߚ ߇ 㧔Fig.2A-C㧕ޔࠩࠗࠧ࠹ࡦᦼߩ߽ߩߪߏߊࠊߕ߆ߒ ߆⷗ࠄࠇߥ߆ߞߚޕࡄࠠ࠹ࡦᦼߩᨴ⦡૕ߪోߊ⷗ࠄ ࠇߥ߆ߞߚޕೋᦼߩ࠺ࠖࡊࡠ࠹ࡦᦼߩᨴ⦡૕ߢߪ ߪߞ߈ࠅߣߒߚ⇣Ᏹࠍ⏕⹺ߔࠆߎߣߪߢ߈ߥ߆ߞߚ ߇㧔Fig.2D,E㧕ޔಝ❗߇ቢੌߒߚ࠺ࠖࠕࠠࡀࠪࠬᦼ ߩ߽ߩߢߪޔ߶ߣࠎߤߩ႐ว 10 ୘ߩᨴ⦡૕ࠍ⏕⹺

ߔࠆߎߣ߇ߢ߈ߚ㧔Fig.2F,G㧕ޕߎߩߎߣߪ⋧หᨴ⦡

૕ห჻߇ಽ㔌ߒߡ߅ࠅޔ৻ଔᨴ⦡૕ߩ⁁ᘒߢሽ࿷ߒ ߡ޿ࠆߎߣࠍ␜ߒߡ޿ࠆޕ

╙৻ಽⵚਛᦼߦߪฦᨴ⦡૕ߪ৻ଔߩ߹߹⿒㆏㕙

ઃㄭߦធㄭߒߡߊࠆ߇㧔Fig.1H㧕ޔ቟ቯߖߕޔᢛ೉

ߔࠆߎߣߪߥ߆ߞߚޕ޽ࠆᨴ⦡૕ߪޔ޿ߕࠇ߆ߩᭂ

ߦᣧߊ߆ࠄ⒖േߒߡߒ߹޿ޔ޽ࠆᨴ⦡૕ߪ╙৻ಽⵚ

⚳ᦼߦߥߞߡ߽⿒㆏㕙ઃㄭߦ஗ṛߒߚ㧔Fig.1J,K㧕ޕ

㊁↢ဳߩࠪࡠࠗ࠿࠽࠭࠽ߢߪᨴ⦡૕ߪਔᭂߦ5ᧄߕ

ߟಽ㈩ߐࠇࠆߩߦኻߒޔߎߩᄌ⇣૕ߢߪ 4㧦6 ߦಽ

㈩ߐࠇߚࠅ㧔Fig.1I㧕ޔ7㧦3ߣ߆8㧦2ߦಽ㈩ߐࠇࠆ

߽ߩ߽޽ߞߚޕߐࠄߦߪ⿒㆏㕙ઃㄭߦ޿ߊߟ߆ߩᨴ

⦡૕߇ᱷߞߡߒ߹޿ޔੑᭂએᄖߩߣߎࠈߦ߽ᨴ⦡૕

߇ಽᏓߒᄙᭂൻߔࠆ߽ߩ߽޽ߞߚ㧔Fig.1L㧕ޕਛߦ ߪޔ╙৻ಽⵚਛᦼ߆ࠄᓟᦼߦ߆ߌߡ৻ଔᨴ⦡૕߇ᆌ ᆂᨴ⦡૕ߦಽⵚߒߡߒ߹ߞߡ޿ࠆߣ⠨߃ࠄࠇࠆ߽ߩ

߽ⷰኤߐࠇߚޕߎࠇࠄߩߎߣߪޔ⋧หᨴ⦡૕߇ࠠࠕ

࠭ࡑߦࠃߞߡㅪ⚿ߐࠇߚੑଔᨴ⦡૕ߣߥߞߡ޿ߥ޿

ߚ߼ߣ⠨߃ࠄࠇࠆޕ

ᷫᢙ╙ੑಽⵚߦ㑐ߒߡߪޔฦᨴ⦡૕߇ᆌᆂᨴ⦡૕

ߦಽ㔌ߒޔ⚦⢩⾰ಽⵚ߇⿠ߎߞߡ⧎☳ዊ⢩ሶ߇૞ࠄ ࠇࠆߣ⸒ߞߚὐߢߪޔ߶߷ᱜᏱߦㅴⴕߒߡ޿ࠆߣ⠨

߃ࠄࠇࠆޕߒ߆ߒޔ╙৻ಽⵚߢᨴ⦡૕߇ᄙᭂൻߒߡ

޿ࠆߚ߼ޔ╙ੑಽⵚߢߪ⚦⢩ౝߩਇ․ቯߩㇱ૏ߢᆌ ᆂᨴ⦡૕߳ߩಽ㔌߇⿠ߎࠅޔ⇣ᢙ૕㧔polyad㧕߇ᒻ ᚑߐࠇߚޕ⇣ᢙ૕߆ࠄᒻᚑߐࠇߚ⧎☳ዊ⢩ሶߪⷙቯ ߩᨴ⦡૕ࠍ᦭ߐߕޔߎߩᄌ⇣૕ߢߪ⒳ሶ߇߶ߣࠎߤ ᒻᚑߐࠇߥ޿ߎߣ߆ࠄޔ߶ߣࠎߤߩ⧎☳ߪ⒤ᕈࠍᜬ ߚߥ޿߽ߩߣ⠨߃ࠄࠇࠆޕ

⸛⺰

AtSPO11-2 ߪ⋧หᨴ⦡૕ߩኻวߦᔅⷐߢ޽ࠆ

SPO11 ㆮવሶߪฎ⚦⩶ߩ DNA topoisomerase VI ߩAࠨࡉ࡙࠾࠶࠻ߣ⋧หᕈ߇޽ࠅޔฎ⚦⩶ߩߘࠇߪ Bࠨࡉ࡙࠾࠶࠻ߣ౒หߢ૕⚦⢩ಽⵚᤨߩDNAⶄ⵾

ߦ㑐ࠊࠆޕ㉂Უ߿᣸⯻ޔ⣄ᬁേ‛ߢߪBࠨࡉ࡙࠾࠶

࠻ㆮવሶߪሽ࿷ߖߕޔSPO11ㆮવሶߪޔᷫᢙಽⵚᤨ

ߩੑᧄ㎮DNA ಾᢿߦ․⇣⊛ߦ㑐ࠊߞߡ޿ࠆޕࠪࡠ

ࠗ࠿࠽࠭࠽ߢߪ3ߟߩAࠨࡉ࡙࠾࠶࠻ߩࡎࡕࡠ࡯ࠣ

㧔AtSPO11-1ޔAtSPO11-2ޔAtSPO11-3㧕ߣ㧝ߟߩ B ࠨ ࡉ ࡙ ࠾࠶ ࠻ ߩ ࡎࡕࡠ ࡯ ࠣ ߇ ሽ࿷ߔ ࠆ ޕ AtSPO11-1 Ⱞ⊕⾰ߪߔߢߦࠪࡠࠗ࠿࠽࠭࠽ߢ߽ᷫ

ᢙಽⵚᦼߩੑᧄ㎮ಾᢿߦ㑐ࠊߞߡ޿ࠆߎߣ߇⍮ࠄࠇ ߡ޿ࠆ4)ޕAtSPO11-3ߪDNAⶄ⵾ߦ㑐ࠊࠆߣߩ⑳

ାࠍᓧߡ޿ࠆ 5)ޕB ࠨࡉ࡙࠾࠶࠻ߩ᦭ήߣᯏ⢻ߩ଻

ሽᕈߣߩ㑐ㅪߪ⥝๧ᷓ޿ޕ㉂Უߢߪᄬࠊࠇߡߒ߹ߞ ߚ௛߈߇ᬀ‛ߢ଻ሽߐࠇߡ޿ࠆߣ޿߁ߩߪޔ৻⷗⍦

⋫ߒߡ޿ࠆࠃ߁ߦᕁ߃ࠆ߆߽⍮ࠇߥ޿߇ޔ㉂Უߩ␲

వߪⶄ⵾ߦ㑐ࠊࠆᯏ⢻ࠍ߽ߞߚ SPO11 ࠍ଻᦭ߒߡ

޿ߚ߇ޔߘߩᯏ⢻ߪㅴൻߩㆊ⒟ߢઁߩㆮવሶߦขߞ ߡઍࠊࠄࠇޔSPO11ߪᷫᢙಽⵚᦼߩᯏ⢻ߩߺࠍ⛽ᜬ ߒߡ޿ࠆߩ߆߽⍮ࠇߥ޿ޕ

AtSPO11-2Ⱞ⊕⾰ߦ㑐ߒߡߪޔߘߩᯏ⢻ߦ㑐ߔࠆ ႎ๔ߪ߹ߛή߆ߞߚ߇ޔBࠨࡉ࡙࠾࠶࠻ߣ⚿วߒᓧ ࠆߣ޿ߞߚᖱႎ߆ࠄޔAtSPO11-3ߣห᭽ߩ௛߈ࠍߔ ࠆߩߢߪߥ޿߆ߣ⠨߃ࠄࠇߡ޿ߚޕߒ߆ߒޔ੹࿁ߩ

⎇ⓥߢatspo11-2ᄌ⇣૕ߢߪ⋧หᨴ⦡૕ߩࡍࠕ࡝ࡦ

(4)

80 Science Journal of Kanagawa University Vol. 17, 2006

ࠣ߇ోߊ⿠߈ߥ޿ߎߣ߇᣿ࠄ߆ߦߐࠇޔAtSPO11-1 ߣห᭽ߦޔᷫᢙಽⵚᦼߩDNAߩੑᧄ㎮ಾᢿࠍߔࠆ ߣ޿߁ߩ߇ߘߩ௛߈ߢߪߥ޿߆ߣផ᷹ߐࠇࠆޕ㉂Უ

߿േ‛ߢߩታ㛎ߦࠃࠅੑᧄ㎮ಾᢿ߇⿠ߎࠄߥ޿ߣ⋧

หᨴ⦡૕ߩࡍࠕ࡝ࡦࠣ߇↢ߓߥ޿ߎߣ߇᣿ࠄ߆ߦߐ ࠇߡ޿ࠆߩߢޔࡍࠕ࡝ࡦࠣߦ⥋ࠆੑᧄ㎮ಾᢿએᄖߩ Ბ㓏ߢ௛޿ߡ޿ࠆน⢻ᕈ߽ቢోߦߪุቯߢ߈ߥ޿߇ޔ AtPO11-2 ߪੑᧄ㎮ಾᢿߦᔅⷐߥㆮવሶߣ⠨߃ࠄࠇ ࠆޕ

AtSPO11-1ߣAtSPO11-2ߩᄌ⇣૕ߪߘࠇߙࠇන

⁛ߩᄌ⇣ߢ⋧หᨴ⦡૕ߩࡍࠕ࡝ࡦࠣ㓚ኂࠍ⿠ߎߔߎ ߣ߆ࠄޔAtSPO11-1Ⱞ⊕⾰߽AtSPO11-2Ⱞ⊕⾰߽

౒ߦሽ࿷ߔࠆߎߣ߇ੑᧄ㎮ಾᢿߦᔅⷐߢޔ ᣇߩㆮ વሶߢ߽ᰳ៊ߔࠆߣੑᧄ㎮ಾᢿߪ⿠ߎࠄߥ޿ߎߣ߇ ࠊ߆ࠆޕฎ⚦⩶ߩDNA topoisomerase VIߪAABB ࠨࡉ࡙࠾࠶࠻߆ࠄߥࠆ྾㊂૕ߢ޽ࠆ߇ޔߎࠇ߆ࠄ㘃 ផߔࠆߣAtSPO11-1ߣAtSPO11-2ߩㆮવሶ↥‛߇

ੑ㊂૕޽ࠆ޿ߪ྾㊂૕ࠍᒻᚑߒߡ޿ࠆน⢻ᕈ߇⠨߃ ࠄࠇࠆޕߒ߆ߒޔ㉂Უ߿േ‛ߢߪ SPO11 ㆮવሶߪ

৻ߟߒ߆⷗ߟ߆ߞߡ޿ߥ޿ޕേ‛߿㉂Უߢߪ૗ࠄ߆ ߩ⵬ഥ࿃ሶ߇௛޿ߡ޿ࠆߎߣ߽⠨߃ࠄࠇࠆ߇ޔ৻ߟ ߩㆮવሶ↥‛߇ᄙ㊂૕ࠍᒻᚑߔࠆߎߣ߽޽ࠆߩߢ޽

ࠈ߁߆㧫޽ࠆ޿ߪੑᧄ㎮ಾᢿ↪ߩ SPO11 ߇ᄙ㊂૕

ࠍᒻᚑߔࠆߣ޿߁ߩߪޔᬀ‛․᦭ߩ⃻⽎ߥߩߢ޽ࠈ ߁߆㧫੹ᓟߩߎߩಽ㊁ߩ⎇ⓥߩ⊒ዷ߇ᦼᓙߐࠇࠆޕ

ᢥ₂

1) Keeney S, Giroux CN and Kleckner N (1997) Meiosis–specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell88: 375-384.

2) ဝౝ⧷↢ (2003) ಴⧘㉂Უߢ᣿ࠄ߆ߦߥߞߚᷫᢙಽ

ⵚᦼߩ⋧หᨴ⦡૕ಽ㈩ࡔࠞ࠾࠭ࡓ. ታ㛎කቇ 21 (No. 5 Ⴧೀ): 700-707.

3) Hartung F and Puchta H (2000) Molecular characterization of two paralogous SPO11 homologues in Arabdiospsis thaliana. Nucleic Acid Res. 28: 1548-1554.

4) Grelon M, Vezon D, Gendrot G and Pelletier G (2001)AtSPO11-1 is necessary for efficient meiotic recombination in plants. EMBO J.20: 589-600.

5) Sugimoto-Shirasu K, Stacey NJ, Corsar J and Rob- erts K (2002) DNA topoisomerase VI is essential for endoduplication in Arabidopsis. Curr. Biol.12: 1782-1786.

6) Azumi Y, Toyama T, Igarashi A and Suzuki H (2001) A sensitive fluorescence in situ hybridization proce- dure applicable to whole stages of male maiosis of Arabidopsis thaliana. Chromosome Sci.5: 1-6.

7) Ross KJ, Fransz P and Jones GH (1996) A light microscopic atlas of meiosis in Arabidopsis thaliana. Chromosome Res.4: 507-516.

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