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Diversity of enzymes in isoflavonoid pathway
Key words: isoflavonoid; P450; phytoalexin; prenyltransferase; soybean.
Tomoyoshi Akashi
Department of Applied Biological Sciences, Nihon UniversityFujisawa, Kanagawa 252–0880, JapanFE   ēĩĴ%ÞÛ7`pe[>X'ƨÜ%ÇĖ$ĕƐ;ö Ʋ85'82`?Zrbt][>X (C6-C3) ĳČ&CoA @LUr%ftZrCoAĳČ&C2 « ţ´vƠįBrHu46İ´ñ87(Ayabe et al. 2010)`pe[>X%'ŲŖñ 3ŘÆ 5&ƢèĩƋ"  2& ľ58 7.Ç &fiņēĩ!'Ʋ@qJQw%ìŐ Īêĺ$>P`pe[>X¿`<>W=sCJu&İ´ñŔ Ĥí©7fiņ!'bUtBr]u3>
P`p\u¿&ƭĒ;ö>P`pe[>X&Çĸ¬éİĩ 5&Ƣè%ƛž$ä¦;ďxÿƲR>M$#!'xƗ&>P`pe[>Xư>P`peuƱ đőřų"İ ōŖ½Ð;ŷƤ&đőřų&ƖÍĹĮ;ƇÓ7JEYrĩƋ"$7"2ľ58 7 (Dewick 1993)
 >P`pe[>X'`pe[>X&Bį=qwrÁ 2 53*ƍĪĆ&ĕƐ;öư¼1Ʊfiņ&>P`pe[>X'İĩ"&H
gmZFwJou%ƛž$ä¦;ď" 5Ʋİ´ñŀŌ'8.!
fiņ;Ċý% ŷ:820Ý,#&ŀŌ%46Ʋxűĺ$`pe[>X5Ÿĺ$`<>W=sCJu&ƭĒ;

7İ´ñƙŖ

ƖÍ ,-µÐ8(Ayabe et al.
2010)Ī%jVrēĩglHEI(Lotus japonicus)!'Ʋ>P`p\u¿ ¼ 1 ēĩ% 7>P`pe[>X& Û

T. Akashi - 1

`<>W=sCJu(vestitol)&İ´ñƙŖƖÍ% ƲƛŽƖÍĀ,-+ %ů7
Šťĺ$ƁĎ $8 (Shimada et al. 2007)85İ´ñƖÍ&Ç 'ƲƙŖŒż"=g[ƚƘ¡îÂ%Á DtwZuE3Ʋ5 1Qu]DƋ&ŉƪ :  7Ã´'HuNuILƘ¡%Á PCR"ĵŉřŪŔ!&ęūƁĎ%46µÐ8 7ưƧĈ 2009ƱŅ'ÄÁv=g[ƚƘ¡îÂ;ëž"ƲƙŖĤí;÷ė% ĻĺƙŖ;öDtwu;«Ʀ7ęūĹĮLDqwZuEĢ;ąƏĲ 7 (Akashi et al. 2003 and 2005a)&òĢ'ēĩcDNA;Èŭų3ƙĝ$#&ĵŉřŪŔ!ĹĮƲĻĺ&ƙŖĤí;öDtwu;İ
©Ïĺ=TN>!±çƲ¬»ƖÍ;µÐ72&!þƀ&ƖÍ;±ç7Ƥ%Ī%Ć¨!
7 ĉŊ!'Ʋƨfiņēĩ!&>P`pe[>Xİ´ñ;±6zİ´ñŔ& ÍƑ©;ŨÒ
7""2%Ʋfiņ&>P`pe[>X&ƫ%ơ:7ÇĖ$bsZrƍŇƙŖ% ŚGE+/?#"059$!;,386
 >P`pe[>X&ŕ90%'®ÍŴ&fiņēĩ%Î¾7 ƲfiņÆ!2HFēĩƲŻÍēĩƲŹÍēĩ!Î¾ ŁƆ8 7(Dewick 1993, Mackova et al. 2006, Reynaud et al. 2005)
ư¼1Ʊ
fiņÆ!'ĪÐ& ƪŦ%7"'Ô$ Ʋû¾ĺ% Û 7 «ÍŴ&=liņ=liÖ(Iris)ēĩ'fiņ%Ě!>P`pe[>X;ŵŋ7" ľ58Ʋ
8.!%20ŉz&ēĩ!Î¾ ŁƆ8 7 >P`pe[>X&ƭĒ'Ʋ>P`pe[>X´ñƙŖ(IFS)%47=qwrÁƍ;^XtCJr©¯ì%46`p\[u46 587 Ʋ&¯ì'fiņ!'CYP93CIa`<gqw&JWDthP450 (P450) ô(šƗvăĿ 2006, Ayabe and Akashi 2006)ư¼2Ʊ
>P`peu'á ş BreCJr@LUpwOĖ&ŬğƙŖ%47¯ì!2-^XtCJ>P`p\[u46 587 (Akashi et al. 2005a)$>P`pe[>X'ƣ58ēĩ%Î¾7&!
9 ƴ Ņ'ƲŔŞĺ%Ʀ8ēĩ İı7>P`pe[>X&İ´ñęĕ;ƁĎƲēĩ&ŔŞƑ©"Īĵ$ĚƈŔ&ñŎ%

&ľſ;ç7Ļĺ!=liÖēĩ&IFS

cDNA&µÐ;ƅ/>P`peuƘœ;Ʈŵŋ7=liÖKkwfu=>qL(Iris germanica)&ÀƬ|Ðđ;ƇÓ(Akashi et al. 2005b)ƲcDNAp>apqw;ñƙĝĹĮŔ;ĲęūĹĮLDq     ¼ 2 fiņēĩ!&>P`peuİ´ñIFS, >P`pe[>X´ñƙŖ;     HID, 2-^XtCJ>P`p\[uŬğƙŖT. Akashi - 2

wZuE%46>P`pe[>XİñĤí;łJuErDtwu;µÐç58Dtwu
HwX7Qu]DƋ(Iris-IFS)'P450! Ʋ
āľ&fiņ&IFS (CYP93CIa`<gqw)"'
ÍŔŞĺ%Ʀ8Ť%Î¾Ʋ&=g[ƚƘ¡&µxí'35Ưňß $ ư¼3Ʊ«ÍŴƪ"Ľě®ÍŴƪ& ØÝ'ŕ15000yÝ¤"øÐ8 7=liÖ&IFS;¶0`?Z
rbt][>Xv`pe[>XŔP450& ÍŔŞĘ;ñƲ³P450& ØÝ;øÐ7"Ʋ=liÖ"fiņ&IFS'3Ýz¤%!%Ø

¼ 3 `?Zrbt][>Xv`pe[>Xİ" øĥ8 5 =liÖ"fiņ ´ñŔ P450 & ÍŔŞĘŔŞĘ' ClastalW&IFS'Ʋ°ü%46Ń¿P450 5ĵ$7Ƒ©; ;ĲƏƥŝ´Ģ!ñƒµ>P`pe[>Xİ´ñęū;ö4%$"øĥ87

IFS Qu]DƋ ƂÌ7ĭƍ¯ì'#ŷ:8 7&9 ƴ fiņ IFS%47¯ì!'Ʋ2-^XtCJ>P`p\[u""2%¥İñĩ& 3-^XtCJ`p\[u;İñ7&¯ì'ƲÁƋ`p\[u& 3 &ğŖ '1%á ó 8 pKBr İƲ&æ=qwrÁ&ƍ" 2 &^XtCJr©

7"Ũ58 7 (Hashim et al. 1990)

¥İñĩ& 3-^XtCJ`p\[uư3 &ğŖ á ó 8æ%µŤ%^XtCJrÁÓ8 İñ7Ʊ&Î¾ Ʋzƃ&¯ìęĕ&Ƅõ&x"$ 7Iris-IFS ;ĹĮśùƙĝgDtPwh;Ĳ

=TN>;ŷ"ƲNADPH Î¾{!`p\[uưnaringeninƱ 5 2-^XtCJ>P`p\[u""2%ÔƝ& 3-^XtCJ`p\[u 4)`peu;İñư¼ 4Ʊ
3-^XtCJ`p\[u&İñ'Ʋfiņ IFS "µęĕ!¯ìŷ:8 7";ł¸ 7fiņ& IFS !'djtKwjVquE"ƗĪĵĺÅĵÓ
%46Ʋ¯ì%ëƩ$ƞ=g[ƚĜÁ µÐ8 7(Sawada et al. 2002, Sawada andAyabe 2005) 85&=g[ƚĜÁ' Iris-IFS !'Î8$Iris-IFS !'ĵ$7ĜÁ¯ì%ơ: 7"ï8ƲæÅĵÓ$#%46ă5% R>M&>P`peu'ķĸ&ƢƲúº%$ 7`

¼ 4 Iris-IFS ĹĮśùƙĝgDtPwh;Ĳ=TN>

<>W@LWtK?u"
T. Akashi - 3ģĻ8 7IFS ;Ĳ8(ĉČ>P`peu;ŵŋ$ēĩ!2İı! 7" ćå87ÑƤ%&4$ƅ/ $8 7 Ʋ8.!&"9śùēĩ!&>P`peuİıƝ'ë2Ʈ $Iris-IFS 'fiņ IFS "ĞƎ7" in vitro !ŕ 20 ƮĤí;łƮ>P`pe[>Xİıū;ł Iris-IFS 'Ʋēĩ&iQeqTD@uKZ=quE%
ĆĲ!7"Ũ58ƲĮ¾śùēĩ!&>P`peuİı;ƅ/ 7 Kkwfu=>qLÀƬ|Ðđ'ƲƮß%ƫ8>P`peu;ŵŋ7&İ´ñ%'ƚ©ƙŖƲO-iSrÁƍŇƙŖƲœƍŇƙŖ&ơ} ïÐ87=liÖēĩ&>P`pe[>XƈŔ&ƙŖ'fiņ&2&"'ĬŮ%Ƒ©"øĥ8ƲIFS Æ&`pe[>XŔƙŖQu]DƋ2āľ&ƙŖ"'ĵ$7nZwD$ĕƐ;ö 7²ūí 7Į¾K
kwfu=>qL|Ðđ& EST ƁĎ;Ƒ1

6Ʋ85&Ƙ¡îÂ&£Ĳ3ęūĹĮLDq

wZuEĢ%46Ʋİ´ñƙŖƖÍ&µÐ;ŷÐ!7

HE'(;,3D'&-A=C>

 Glyceollin 'ƲbUtBr]uƭĒ% C5 >Pbsu« §Ʋįī@wUr;ãñR>M&ž$`<>W=sCJu!7ư¼ 5Ʊ
Glyceollin 'bsZrÁ&§Ť"&æ&į©Ėà&Ɣ 5Ʋ3 ŉƪ&ĵí(glyceollin I, II, III) Î¾7&İ´ñŔ&ƙŖƖÍ&Ç '!%µÐ8 ƲŜƌæª&R>MĪĆ&¯ì;ôƙŖ% 'ƖÍscr&ľſ'$ Ī%bUtBr]u(glycinol)& 2 " 4 %bsZrÁ;ƍŇ7ƙŖ'Ʋ
1980 Ý%Ĥí Ĕ8Ʋglyceollin İ´ñƙŖ&~!¹xŲŖ%Õ¾7"ł¸8 
7 ƲÑ'|ă!
8HOO

654'OOHODaidzein

OHHO

4HO
2

6aO

OOHO

3OOH

G2DT

11aO

GlucoseOOH
2-Dimethylallylglycinol

9 OH(-)-Glycinol[(-)-3,9,6a-trihydroxypterocarpan]

OOOH

Dimethylallyltransferase (DT)

OCyclases[P450s]OPPMEP pathway

OHGlyceollin III2OHGlyceollin IIG4DT
HO4O
OHO
OH4-Dimethylallylglycinol

OOOHOOHGlyceollin I

Soybean phytoalexin¼ 5 R>M&EqNAqu&İ´ñŜƌ

T. Akashi - 4 ŏũ5'bUtBr]u*&bsZrÁƍŇ¯ìWH`?twr´ñŔ& homogentisate

phytyltransferase (HPT)"¯ì ƪ 7" 5ƲHPT =g[ƚƘ¡;2"%R>M EST V
wQcwL;ĔŗƲ50Ưz&µxí;ł 3 &źƘ¡ưPT1, PT2, PT3Ʊ;ƕó& PT2 'R>MĶĸų ðĐřŪ& EST p>apqw!â ĹĮ6ƲƙĝĹĮŔ;ĲęūƁĎ%46 glyceollin I &İ´ñ%ơ:7(-)-glycinol 4-dimethylallyltransferase
ưG4DTƱ!7" : (Akashi et al. 2009)&æR>M&G[hƘ¡ ă5 %8
" 5VwQcwL;ßĔŗƲþ% 3 &djtE(PT4, PT5, PT6);ſ
88;ƙĝ!ĹĮ ęūƁĎ;ŷ"Ʋ
PT6 ' glyceollin II, III &İ´ñ%ơ:7(-)-glycinol
2-dimethylallyltransferaseưG2DTƱ!7" : . PT4, PT5 '88>P`pe[>X&DiLUtwr­)>P`peu;bsZr&=DNbQw%7" :  fiņ&Dpp(Sophora flavescens) 5'`p\[u& 8 %bsZrÁ;ƍŇ7ƙŖƖÍ(SfN8DT-1, SfN8DT-2) µÐ8 7(Sasaki et al. 2008)ŔŞĘz!' PT2, PT3, PT4, PT5,PT6 ' SfN8DT "'¢&DswX%Ö 7R>M!'Ƒ©&Ɠň!>P`pe[>X;Á
Ƌ%7bsZrÁƍŇƙŖ İƲÇĖ$ęū;ö4%$"ïÐ87=DNbQw&ĕƐ;Ġ17Qu]DƋĕƐ%Ű· ö8Ʋæ³Ƙ¡&ĞƎ3ÅĵÓ%46ă5 %¼ 6 ēĩ&bsZrÁƍŇƙŖ& ÍŔŞĘŔŞĘ' ClastalW ;ĲƏƥŝ´Ģ!ñ

1<4:ƧĈÊ 2009ƳfiņjVrēĩ&G[hîÂ"\>AqPwL;£ĲÉħĩİ´ñƖÍ&µÐƳ`<rfJ=Ʋ45: 881-886.
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