Ehime University Institutional Repository

BOGDAN, David R.

English

Integrating the Moodle LMS with Classroom Instruction into a "Blended" Learning Environment : The Beginnings of a Case Study

岡村, 健一

山本, 夕奈

水本, 憲枝

田内, 広子

長尾, 秀夫

Japanese

A Study to the Way of Thinking of Multidisciplinary Habilitation Staffs (No.16) : The present and review for Developmental Support of Very Low Birth Weight Children

東, 賢司

愛媛大学教育学部紀要　第57巻　207 ～ 212　2010
207
はじめに
昨年，「馬衡・唐蘭編 未刊本『甲骨刻辞』調査記録」
を脱稿したあとに，韓国人の李鐘淑氏と葛英會氏が著書
であり，北京大学中国考古学研究中心・北京大学考古文
博学院が編集した『北京大学珍蔵甲骨文字』（上海古籍
出版社，2008年11月）を入手した（以下「北珍」と省略）。
この著録は，北京大学震旦古代文明研究中心学術叢書特
刊として発行されている。このセンターは元々北京大学
古代文明研究中心とされており，1999年５月に設置さ
れている。2003年１月に和震旦文教基金会と協議が行
われ，現在のセンターとなっている。
この北珍であるが，序文から『甲骨刻辞』の文言を見
ることはできない。序文などから，北京大学の甲骨文の
集録の歴史が分かってくる。本論では，昨年明らかにで
きなかった北京大学の甲骨片について補足を行いたい。
１．北京大学の甲骨文字
表１　北京大学の甲骨片の来歴
年代 数量 来歴 帰属単位
1922 463 霍保禄蔵 北京大学研究所国学門
1929 1200 容庚購買 燕京大学国学研究所
1936 G.A.H贈
1950 690
1951 29
1954 5
  
北京大学に現存する甲骨文字は，北京大学と燕京大学
の両校が旧蔵する資料を基礎としている。1952年に両
校が収蔵する甲骨が一つになり，北京大学歴史系考古教
研室の管理となり，1993年に北京大学考古與藝術博物
館が建設されると，その資料室の珍蔵とされた。しかし，
収蔵の記録は非常に簡単である。北京大学考古與藝術博
物館によると，大学への寄贈者等は，表１のようになる。
1963－1966年に閻文儒が採拓を行い『甲骨目録』を
編集したが，甲骨片の総数は，3022片となっている。
また，甲骨文字の専著には，北京大学蔵の甲骨が集録
されている。これらをまとめると以下のようになる。
Ａ．殷契卜辞
1933年，容庚が燕京大学に購買した1200片の甲骨か
ら874片を選定し，哈佛燕京学社印行として出版した。
Ｂ．殷契佚存
1933年，商承祚の著作。北平孫氏の甲骨墨本193紙中，
34紙に北京大学研究所国学門の甲骨拓本が含まれる。
Ｃ．殷虚書契続編
1933年，羅振玉の著作。北京大学国学門所蔵甲骨を
183片集録する。
Ｄ．戦後南北所見甲骨録
1951年に胡厚宣が編著。巻二の「南北師友所見甲骨録」
の模本は，大部分が北京大学の所蔵である。
Ｅ．北京大学国学門蔵殷虚文字考釋
1980年に厳一泙が編著。模本を266片集録。
Ｆ．甲骨文合集
1982年に完成した甲骨文の代表著作であるが，北京
大学考古與藝術博物館の甲骨を1149片集録している。
序文にも記されているように，北京大学には多くの甲
骨片が集録されているにもかかわらず，分類や読解等の
総合的な研究がされてこなかった。また，1966年の拓
本製作から4片の甲骨が行方不明である。更に偽刻や無
字のものも含まれるために，合計数は2929片となる。
　
馬衡・唐蘭編 未刊本『甲骨刻辞』調査記録（補）
（書写書道教室）　　東　　　賢　司
Search Records of Unpublished “Jia Gu Ke Ci” edited by Ma Heng and Tang Lan
（Supplementation）
HIGASHI Kenji
（平成22年６月５日受理）
東　　　賢　司
208
２．『甲骨刻辞』との比較
未刊本『甲骨刻辞』と『北珍』を比較した表を下に
記す（表２）。北京大学に現存する甲骨は154片である。
また，昨年の調査では，未刊本『甲骨刻辞』380片の原
拓中，発行済みの著録に集録されていることが確認でき
たのは241件であったが，新たに61件の所在を明らかに
できた。残りの甲骨拓本78片は，未確認である。
表２　『甲骨刻辞』と『北京大学珍蔵甲骨文字』の比較
【凡例】
１．「正反」「期」「内容分類」については，『北京大学珍蔵甲骨文字』から採録した。
２．著録等の略号は以下のとおり。刻辞…『甲骨刻辞』(私蔵本)，北登…北京大学考古與藝術博物館登録記号，続…羅振玉『殷
虚書契続編』1933年，佚…商承祚『殷契佚存』1933年，南…胡厚宣『戦後南北所見甲骨録』1951年，考釈…嚴一萍『北京大學
國學門藏殷虚文字考釋』1980年，合集…郭沫若『甲骨文合集』1977－1982年，考 …中国社会科学院考古研究所考 室， 初
下…黄濬『 中片羽初集(下)』1935年，文捃…甲骨文捃(曽毅公)
３．『北京大学珍蔵甲骨文字』に伏される番号は，「北京大学所蔵甲骨文字著録重見表」に基づいている。
４．「分類」覧の数字は，１農事，２田猟，３祭祀，４戦争，８王事，９天象・気象，10干支・暦数，11卜法，12其他である。
５．「期」は，第Ⅰ期から第Ⅴ期までとした。
刻辞 北珍 北登 続 佚 南 考釈 合集 其他 分類 期
10 2052 8.036 　 　 2.166 4.15.2 3531 　 12 Ⅰ
12 2822 8.0304 　 　 2.187 2.06.4 24253
41071
12
Ⅱ
13 1006 8.0249 　 　 　 　 285 　 8 Ⅰ
14 1249 8.0252 4.37.9 　 　 3.09.4 34982 　 8 Ⅳ
14 1250 8.0230 4.39.4 　 　 3.22.4 34844 　 8 Ⅳ
15 1247 8.0243 4.40.4 　 　 3.05.3 34868 　 8 Ⅳ
17 998 8.0466 　 　 　 　 　 　 8 Ⅰ
18 994 8.0125 　 　 　 2.10.3 　 　 8 Ⅰ
19 1012 8.0210 4.47.3 　 　 4.18.2 16641 　 8 Ⅰ
22
995
(正反)
8.0251
(正反)
4.48.4
(正)
1.11.3
(正)
16895
(正反)
考 465
(正) 8
Ⅰ
24 1226 8.0241 4.48.10 　 　 2.13.2 31320 　 8 Ⅲ
27
2044
(正反)
8.0411
(正反)
2.80(正)
2.81(反)
4.05.4(正)
4.05.5(反)
17305
(正反) 11
Ⅰ
29 2043 8.0121 　 　 2.132 4.27.4 16942 考 64 11 Ⅰ
30 1376 8.0232 　 　 2.264 1.23.4 41934 　 8 Ⅴ
31 1379 8.0246 　 　 　 3.03.4 39127 考 482 8 Ⅴ
32 1342 8.0228 　 　 　 2.14.3 　 考 403 8 Ⅴ
34 1339 8.0234 6.5.1 　 　 2.12.1 39318 　 8 Ⅴ
35 1330 8.0220 　 　 　 　 　 　 8 Ⅴ
36 1419 8.0218 　 　 　 3.06.1 　 　 8 Ⅴ
39 1366 8.0206 6.4.3 　 　 4.33.3 39051 　 8 Ⅴ
40 1388 8.0229 　 　 　 3.01.1 39058 　 8 Ⅴ
55 1328 8.0255 6.2.8 　 　 3.04.4 39110 　 8 Ⅴ
56 1371 8.0097 　 　 　 3.06.2 　 　 8 Ⅴ
57 1378 8.0245 6.3.10 　 　 2.29.1 39369 　 8 Ⅴ
58 1398 8.1003 　 　 　 　 　 　 8 Ⅴ
馬衡・唐蘭編 未刊本『甲骨刻辞』調査記録（補）
209
61 1362 8.0005 　 　 　 3.08.3 37902 　 8 Ⅴ
62 142 8.0295 　 　 2.99 2.24.3 4917 　 3 Ⅰ
63 1174 8.0276 　 　 　 　 　 　 8 Ⅱ
68 1788 8.0021 　 　 2.125 4.32.1 41595 　 10 Ⅰ
70
1890
(正反)
8.0309
(正反)
4.5.5
(正)
2.28(正)
2.29(反)
3.20.1(正)
3.20.2(反)
5579
(正反) 11
Ⅰ
72 1437 8.0051 　 　 2.174 3.19.4 3928 　 9 Ⅰ
74 1276 8.0226 　 　 2.258 3.08.4 41907 　 8 Ⅴ
76 1273 8.0235 　 　 2.262 3.24.2 38892 　 8 Ⅴ
77 1283 8.0225 　 　 　 3.07.3 　 　 8 Ⅴ
79 1279 8.0223 　 　 　 　 　 　 8 Ⅴ
80 1263 8.0211 　 　 　 　 39173 　 8 Ⅴ
82 1254 8.0222 　 　 2.263 1.13.2 38885 　 8 Ⅴ
83 1278 8.0237 　 　 　 4.22.2 　 　 8 Ⅴ
85 1294 8.0238 　 　 2.259 4.33.1 41906 　 8 Ⅴ
86 1253 8.0221 　 　 　 2.11.4 　 　 8 Ⅴ
88 1286 8.0446 　 　 　 　 　 　 8 Ⅴ
91 1293 8.0012 3.19.8 　 　 2.03.2 36881 　 8 Ⅴ
94
1475
(正反)
8.0417
(正反) 188
3.27.1(正)
3.17.2(反)
12807
(正反) 9
Ⅰ
96 1481 8.0334 　 　 2.193 3.22.1 11998 　 9 Ⅰ
100
54
(正反)
8.0258
(正臼)
4.7.2
(正)
2.16(正)
2.15(臼)
3.19.1(正)
3.19.2(臼)
10199
(正臼) 2
Ⅰ
106
1476
(正反)
8.0370
(正反)
4.33.6
(正)
2.36(正)
2.37(反)
3.28.1(正)
3.28.2(反)
17680
(正反) 考 316 9
Ⅰ
107 1607 8.0049 4.18.2 　 　 2.28.3 29933 　 9 Ⅲ
107-3 2161 8.0311 　 　 2.159 4.05.3 17870(反) 12
Ⅰ
108 1441 8.0418 4.23.6 　 　 3.25.2 12178 　 9 Ⅰ
109 1591 8.0327 4.17.8 　 　 3.19.5 24770 　 9 Ⅱ
110 1503 8.0331 　 　 2.172 3.26.1 12040 　 9 Ⅰ
112
1536
(正反)
8.0316
(正反)
4.24.5
(正)
3.27.3(正)
3.27.4(反)
12873
(正反)
考 298
(正) 9
Ⅰ
114 1519 8.0410 　 　 　 1.20.4 　 　 9 Ⅰ
117 1623 8.0399 4.14.8 　 　 2.29.4 38201 　 9 Ⅴ
120 1572 8.0126 　 　 　 1.12.3 1407 　 9 Ⅰ
121 1774 8.0148 　 　 　 2.02.2 24935 　 10 Ⅰ
122 2509 8.0053 　 　 　 　 　 　 12 Ⅰ
167 534 8.0162 2.4.2 　 　 2.14.2 38576 　 3 Ⅴ
168 543 8.0177 　 　 2.238 1.28.3 　 　 3 Ⅴ
170 375 8.0157 　 　 　 1.31.3 　 　 3 Ⅱ
171 587 8.0156 　 　 　 4.22.1 　 　 3 Ⅴ
172 569 8.0181 　 　 　 1.31.4 39474 　 3 Ⅴ
174 586 8.0163 2.10.8 　 　 1.20.3 38389 　 3 Ⅴ
175 571 8.0178 　 　 2.244 1.29.3 41889 　 3 Ⅴ
177 573 8.0165 　 　 2.243 1.30.2 41151 　 3 Ⅴ
東　　　賢　司
210
179 598 8.0180 　 　 2.242 1.31.1 　 　 3 Ⅴ
182 525 8.0174 2.5.5 192 　 2.13.4 35376 　 3 Ⅴ
183 492 8.0147 1.22.10 　 　 1.20.2 35737 　 3 Ⅴ
184 603 8.0159 　 　 　 1.27.4 　 　 3 Ⅴ
187 602 8.0183 　 　 　 1.28.2 　 　 3 Ⅴ
188 658 8.0045 1.26.4 　 　 1.14.4 35915 　 3 Ⅴ
189 675 8.0063 6.7.4 861 　 1.04.1 36156 　 3 Ⅴ
190 659 8.0443 1.26.2 　 　 1.05.3 35947 　 3 Ⅴ
193 680 8.0209 　 　 2.232 1.05.2
4.30.3 35934 3
Ⅴ
194 692 8.0047 1.24.6 　 　 1.06.4 35850 　 3 Ⅴ
195 693 8.0197 　 　 2.247 1.34.4 37350 　 3 Ⅴ
196 731 8.0187 2.26.9 　 　 1.36.1 37070 　 3 Ⅴ
197 715 8.0205 2.16.5 　 　 1.36.4 37038 　 3 Ⅴ
199 728 8.0199 　 　 　 1.19.1 　 　 3 Ⅴ
201 732 8.0198 2.26.10 　 　 1.35.1 37211 　 3 Ⅴ
202 677 8.0025 1.43.4 　 　 1.25.4 36322 　 3 Ⅴ
204 717 8.0195 　 　 　 1.38.1 37280 　 3 Ⅴ
207 2885 8.0274 　 　 　 2.16.1 　 　 12 Ⅴ
210
190
(正反) 8.0076 1.5.5 　 2.51 4.11.3 14313 　 3
Ⅰ
211 74 8.0280 3.40.1 　 　 2.21.4 10939 　 2 Ⅰ
212 187 8.0069 1.2.1 　 　 1.33.4 358 　 3 Ⅰ
213 183
8.0313
　
1.52.3 　 2.63,2.64 1.39.3
1.39.4 7919 3
Ⅰ
214
1453
(正反)
8.0067
(正反)
1.27.9
(正)
2.42(正)
2.44(反) 1.15.4
12495
(正反) 9
Ⅰ
215 189 8.0185 2.26.11 　 　 3.26.4 22434 　 3 Ⅰ
216 201 8.1298 　 　 　 　 16263 　 3 Ⅰ
218 456 8.0192 　 　 　 1.34.3 　 　 3 Ⅲ
221 168 8.0353 4.34.2 　 　 3.30.4 4641 　 3 Ⅰ
223 361 8.0277 2.3.10 　 　 2.04.3 25155 　 3 Ⅱ
225 346 8.0201 2.25.5 　 　 1.15.1 25974 　 3 Ⅱ
226 209 8.0068 2.23.9 180 　 1.32.2 6035 　 3 Ⅰ
229 269 8.0082 1.36.4 145 　 3.10.4 14548 　 3 Ⅰ
230
1149
(正反)
8.0132
(正反)
4.23.1(正)
4.23.2(反) 8
Ⅰ
231
24
(正反)
8.0079
(正反)
1.41.6
(正)
153
(正) 1.22.2(正)
10130
(正反) 1
Ⅰ
233 453 8.0143 　 　 　 2.01.1 15264 　 3 Ⅲ
235 28 8.0066 1.45.4 46 　 1.36.2 10117 考 59 1 Ⅰ
236 29 8.0081 1.50.4 　 　 3.17.4 10102 　 1 Ⅰ
237 267 8.0191 2.17.4 　 　 1.33.3 15898 　 3 Ⅰ
238 211 8.0202 　 　 　 1.37.3 2941 　 3 Ⅰ
239 206 8.0108 　 　 　 2.11.2 2963 考 467 3 Ⅰ
240 207 8.0028 1.39.3 181 　 1.08.4 331 　 3 Ⅰ
馬衡・唐蘭編 未刊本『甲骨刻辞』調査記録（補）
211
244a 2378 8.0450 　 　 　 　 805 　 12 Ⅰ
244b 2678 8.0441 　 　 　 　 　 　 12 Ⅰ
246b 2494 8.0394 　 　 　 　 　 　 12 Ⅰ
248 159 8.0013 1.45.7 　 　 4.30.4 1972 　 3 Ⅰ
250 150 8.0468 1.47.1 159 2.19 1.16.1 3467 　 3 Ⅰ
252 172 8.0428 　 　 　 1.39.1 　 　 3 Ⅰ
254 329 8.0083 　 　 2.55 3.29.2 17358 　 3 Ⅰ
255
158
(正反)
8.0322
(正反)
1.39.1
(正)
2.1(正)
2.2(反)
1.24.1(正)
1.24.2(反)
2410
(正反) 3
Ⅰ
256 167 8.0122 　 　 　 4.12.2 15092 　 3 Ⅰ
258 1659 8.0036 　 　 　 　 　 　 10 Ⅰ
260
2185
(正反)
8.0376
(正反) 11
Ⅰ
261 2195 8.0379 　 　 　 　 　 　 12 Ⅰ
263 1098 8.0052 　 　 　 3.29.1 　 　 8 Ⅰ
269 175 8.0054 2.19.3 413 　 1.26.2 22543 初下
30.2 3
Ⅰ
274 301 8.0061 1.46.4 10 　 4.08.3 1672 　 3 Ⅰ
275 462 8.0070 1.4.5 140 　 1.12.2 　 　 3 Ⅳ
275b 2236 8.0382 　 　 　 　 　 　 12 Ⅰ
277 467 8.0030 　 　 2.237 4.24.3 38301 　 3 Ⅴ
278
2388
(正反)
8.0377
(正反)
2.3(正)
2.4(反)
1.06.1(正)
1.06.2(反)
1628
(正反) 11
Ⅰ
279
786
(正反)
8.0057
(正反)
1.17.6
(正)
2.90(正)
2.91(反) 1.18.1(正)
1774
(正反) 4
Ⅰ
280
1891
(正反)
8.0459
(正臼) 2.23(正) 1.39.2(正)
17548
(正臼) 11
Ⅰ
281 1892 8.0461 5.11.5 160 　 4.09.4 17568 　 11 Ⅰ
282
1888
　
8.031
8.0190 2.30
3.29.4
　
17600 11
Ⅰ
283
2020
(正反)
8.0117
(正反)
2.82(正)
2.83(反)
3.11.1(正)
3.11.2(反)
4307
(正反) 11
Ⅰ
284 2 8.0039 3.47.1 　 　 4.18.4 4104 　 1 Ⅰ
288
2367
　
8.0090 　 　 2.43 1.25.1 19116
40770
12
Ⅰ
290 2201 8.0432 　 163 　 4.07.1 19558 　 12 Ⅰ
291c 107 8.0151 　 　 　 4.21.1 37420 　 2 Ⅴ
291d 2226
　
8.0096 　 　 2.254 4.25.2 36385
41897
12
Ⅰ
291e 2110 8.0072 　 142 2.97 4.08.2 4274 　 12 Ⅰ
293 817 8.0462 4.30.1 527 　 3.14.2 6459 　 4 Ⅰ
294 2074 8.0123 　 　 2.74 4.24.4 4577 　 12 Ⅰ
295 1106 8.0285 3.39.2 　 　 2.20.5 2642 　 8 Ⅰ
308 852 8.0458 　 　 2.148 4.17.1 3380 　 4 Ⅰ
309 1132 8.0355 　 　 2.149 4.15.1 5465 　 8 Ⅰ
311 792 8.0264 3.7.2 13 　 2.25.2 6196 　 4 Ⅰ
東　　　賢　司
212
312 763 8.0077 1.10.3 　 　 1.10.3 6168 　 4 Ⅰ
313a 566
　
8.0171 　 　 2.24 4.19.1 38596
41885
3
Ⅴ
313e 1158 8.0046 　 　 2.103 2.21.3 54 文捃935 8 Ⅰ
313f 1297 8.0236 　 　 2.25 4.29.3 41751 　 8 Ⅴ
318 806 8.0383 　 　 　 　 　 　 4 Ⅰ
319 208 8.0425 　 　 　 3.05.2 6612 　 3 Ⅰ
320 767 8.0261 5.25.9 　 　 2.26.2 6761 　 4 Ⅰ
321 858 8.0426 　 　 2.98 4.20.4 6562 　 4 Ⅰ
322 859 8.0460 　 　 2.96 4.12.4 7705 　 4 Ⅰ
323 853 8.0339 　 141 　 4.09.2 6774 　 4 Ⅰ
324 1186 1052 　 　 　 　 37857 　 8 Ⅱ
326 826 8.0136 　 　 　 2.16.3 16261 　 4 Ⅰ
327 868 8.0281 　 　 2.95 2.22.4 492 　 4 Ⅰ
※この研究は，基盤研究（B）海外学術調査「故宮博物院に収蔵される甲骨文の来源調査－未刊本『甲骨刻辞』の解
読を通して―」（課題番号：19401028）及び平成21年度財団法人髙梨学術奨励基金研究助成（研究題目：故宮博物院
に収蔵される甲骨片流転の遍歴調査研究）の成果の一部である。


Search Records of Unpublished“ Jia Gu Ke Ci" edited by Ma Heng and Tang Lan

RABBANY, A.B.M. Golam

MIZUTANI, Fusao

CHIKAIZUMI, Sojiro

Introduction
Essential oils are volatile substances that evolve from
plant tissues as secondary metabolic products. These
secondary plant products are of importance in biological
and ecological aspects in plants (Croteau, 1987; Tigney et
al., 1991). We revealed that some essential oils inhibited
ethylene biosynthesis in apple fruit and peach seed tissues
(Rabbany and Mizutani, 1996). The enzymes involved in
ethylene biosynthesis are ACC synthase and oxidase,
which catalyze the conversion of S-adenosylmethionine
(SAM) to ACC and ACC to ethylene, respectively. Here,
we report effects of essential oils on in vitro activities of
ACC synthase and ACC oxidase obtained from wounded
winter squash mesocarp tissues.
Materials and Methods
Plant materials
Winter squash fruit (Cucurbita maxima Duch. cv.
Ebisu) were obtained from a local market and stored at 12
℃ until use. Plugs (10mm in diameter) were prepared
from the mesocarp of fruit and cut into 2 mm thick slices.
The slices were incubated in a plastic chamber on a moist
paper towel for 16 hr at 24℃ under 90% relative
humidity.
Essential oils
Three groups of essential oils were used: limonene as a
hydrocarbon, linalool as an alcohol and citral, citronellal,
1-octanal, 1-nonanal and 1-decanal as aldehydes. All of
the essential oils were purchased from Wako Pure
Chemical Industries Ltd.
ACC synthase extraction and assay
The mesocarp tissues were homogenized with a chilled
pestle and mortar in 2ml/g tissue of extraction buffer (A)
containing 0.1M 3-[4-(2-hydroxyethyl)-1-piperazinyl] pro-
panesulfonic acid (EPPS) buffer (pH8.5), 5mM 2-
mercaptoethanol, 5μM pyridoxal phosphate (PLP) and 2%
(w/w) insoluble polyvinylpyrrolidone (Tokyo Chemical
Industry Co. Ltd.). The homogenate was centrifuged at
8000 x g for 20min at 0℃. The supernatant was passed
through a column of Sephadex G-25 (PD-10; Pharmacia,
Uppsala, Sweden) that had been equilibrated with the
extraction buffer (A). The macromolecular fraction was
collected and used as a crude enzyme extract. All
procedures were carried out at 2℃. The enzyme activity
was assayed in a sealed 70ml test tube, which contained
20mM EPPS, 200μM SAM, 1.6μM PLP and 0.6ml of the
enzyme extract. A given amount (0.25, 0.5, 1.0, 2.5μl) of
oils was applied with micro syringes (1.0 and 10μl full
scale) to the interior wall of the test tube and the outer
wall was heated with a hair dryer for vaporizing the
essential oils and then cooled. After adding the enzyme
Effect of Essential Oils on In Vitro Activities of
1-Aminocyclopropane-1-carboxylate (ACC) Synthase and ACC Oxidase
from Winter Squash Mesocarp
A. B. M. Golam RABBANY１，２）, Fusao MIZUTANI１）and Sojiro CHIKAIZUMI１）
Abstract
In vitro experiments were carried out to investigate the effect of essential oils on 1-aminocyclopropane-1-carboxylate
(ACC) synthase and ACC oxidase by using crude enzyme extracts from wounded mesocarp tissues of winter squash
(Cucurbita maxima Duch. cv. Ebisu). Enzyme activities were assayed in a closed vessel including essential oils in its
head space. All of the essential oils tested inhibited ACC synthase activity. A 50% inhibition was obtained with
limonene and linalool at 28 and 25μl/l (liquid volume in the head space), respectively. Greater inhibition was found
with aldehyde group essential oils. Citral, 1-octanal, 1-nonanal and 1-decanal inhibited ACC synthase activity by 50 to
78% and citronellal by 78 to 93% at their applied amounts ranging 3.6 to 36μl/l. On the other hand, the essential oils
did not inhibit ACC oxidase activity. However, aliphatic aldehydes such as 1-octanal, 1-nonanal and 1-decanal caused
a non-enzymatic breakdown of ACC into ethylene.
Key words: ACC synthase, ACC oxidase, aliphatic aldehydes, non-enzymatic breakdown of ACC
Received April 1, 2010
Accepted July 30, 2010
１）Laboratory of Citriculture, Faculty of Agriculture, Ehime
University, ３‐５‐７ Matsuyama, Ehime Prefecture, ７９０‐
８５６６, Japan
２）Present address: １６４６ W. Davenport St. Apt ＃８, Rhine-
lander, Wisconsin, WI５４５０１, USA
愛媛大学農学部紀要（Mem. Fac. Agr., Ehime Univ.）
５５：１－６（２０１０）
１
solution with a hypodermic syringe, the test tube was
incubated at 30℃ for 30min with reciprocal shaking (40
strokes/min). The reaction was stopped by addition of 0.1
ml of 20mM HgCl2. ACC formed from SAM by ACC
synthase was assayed by the method of Lizada and Yang
(1979).
ACC oxidase extraction and assay
The mesocarp tissues of winter squash fruit were
homogenized with a pestle and mortar in 2ml/g tissue of
of extraction buffer (B) containing 0.1M Tris-HCl (pH
7.4), 10% (v/v) glycerol and 30mM Na-ascorbate. The
homogenate was centrifuged at 10,000 x g for 15min at 0
℃. The supernatant was used as a crude extract. All
procedures were carried out at 2℃. The enzyme activity
was assayed in a sealed 55-ml plastic syringe, which
contained 0.2M 3-morpholinopropane-1-sulfonic acid
(MOPS) (pH6.7), 10% (v/v) glycerol, 30mM Na-
ascorbate, 50μM FeSO4, 1mM ACC, 0.15% Triton X-100,
14mM NaHCO3, and 0.2ml of the crude extract in a total
volume of 1.5ml and 20% CO2 in the head space.
Essential oils (0.2, 0.4, 0.8, 2.0μl) were injected into the
head space and vaporized as mentioned above. The
syringe was incubated at 30℃ with reciprocal shaking (60
strokes/min). After 30min incubation, ethylene content in
the syringe was determined by using a gas chromatograph
equipped with an activated alumina column.
Results
Effects of various essential oils on ACC synthase activities
ACC synthase activities as affected by the essential oils
are shouwn in Fig. 1. All of the essential oils inhibited
ACC synthase activity. This inhibition occurred at a very
low amount (3.6μl/l) and increased with their increasing
amounts. Limonene, a hydrocarbon essential oil, inhibited
ACC synthase activity by 50% at 28μl/l, whereas linalool,
an alcohol essential oil, inhibited the activity by 42 to
52% over the range from 3.6 to 36μl/l. Aldehyde essential
oils such as citral, citronellal, 1-octanal, 1-nonanal and 1-
decanal greatly inhibited ACC synthase activity. Citronel-
lal was most effective in the inhibition of ACC synthase
activity among the essential oils tested. Citral inhibited
ACC synthase activity by 50 to 78% over 3.6 to 36μ/l,
whereas citronellal inhibited 78 to 93% with the same
dosages. 1-Octanal, 1-nonanal and 1-decanal were similar
in the inhibition of ACC synthase activity. Among these
three aliphatic aldehydes, 1-octanal gave the most pro-
nounced inhibition.
Effects of various essential oils on ACC oxidase activities
Essential oils had no effect on ACC oxidase activity,
except for three aliphatic aldehyde essential oils (Fig. 2).
The incubation of the enzyme with different amounts of 1
-octanal, 1-nonanal and 1-decanal caused a significantly
higher rate of ethylene production with their increasing
amounts. At 36μl/l of C8, C9 and C10 aldehyde essential
oils, ethylene production rates were 2.8-, 18.2- and 12.9-
fold over the control, respectively (Fig. 2).
Non-enzymatic breakdown of ACC by aliphatic aldehydes
The stimulation of ethylene production might be
associated with non-enzymatic breakdown of ACC in the
presence of these aliphatic aldehydes or the breakdown of
themselves. This speculation was tested by monitoring
ethylene evolution in enzyme-free and ACC-free reaction
mixture with 1-nonanal. As shown in Fig. 3, a high
ethylene production occurred in the assay mixture even
without the enzyme extract, and there was no ethylene
evolution in ACC-free mixture.
Discussion
Rabbany and Mizutani (1996) previously reported that
essential oils, especially aldehyde types, suppressed the
increase of ACC content and ethylene production in the
assay by using excised young peach seeds. The young
peach seed exhibits a rapid increase in ACC content and
ethylene production just after taken from the pit (Jerie and
Chalmers, 1976). The present results confirmed that the
suppression is due to the inhibition of ACC synthase
activity. ACC synthase is a PLP-requiring enzyme (Boller
et al., 1979; Hyodo et al., 1983; Yoshii and Imaseki,
1981; Yu et al., 1979). Most of the PLP-linked enzymes
have a lysine residue in their active site, where the ε-
amino group of lysine makes a Schiff base with aldehyde
group of PLP coenzyme. In our study the aldehyde
essential oils were found to be most inhibitory to ACC
synthase. If they form a Schiff base with the ε-amino
group of lysine residue, the formation of the aldemine
linkage between PLP and the ε-amino group of lysine
residue of the enzyme might be hampered, which
consequently results in the inhibition of ACC synthase
activity. Therefore, further kinetic studies will reveal
whether such a competition between the aldehyde
essential oils and PLP operates during ACC synthase
action. On the other hand, concerning other types of
essential oils such as limonene and linalool, the
mechanism in which they inhibit ACC synthase activity
remains to be elucidated.
We also observed that the rate of non-enzymatic
conversion of ACC to ethylene increased with increasing
numbers of carbon in aliphatic aldehydes, whereas
aromatic aldehydes showed no effect (data not shown).
Earlier reports described that simple aldehydes such as
methional (Beauchamp and Fridovich, 1970) and propanal
(Baur and Yang, 1969) produce ethylene non-
enzymatically. However, these aldehydes themselves
convert to ethylene. Non-enzymatic conversion of ACC to
ethylene has been reported previously under various
conditions. Divalent metal ions, free radicals and
lipoperoxides (Yang and Hopffman, 1984; McRae et al.,
1983; Kaaceperska and Kubacka-Zehalska, 1993) facilitate
the non-enzymatic conversion of ACC to ethylene. There
is no information on the non-enzymatic conversion of
ACC to ethylene caused by aliphatic aldehydes.
２ A. B. M. Golam RABBANY, Fusao MIZUTANI and Sojiro CHIKAIZUMI
Fig. 1 Effect of essential oils on in vitro activity
of ACC synthase obtained from wounded
winter squash mesocarp tissues. Dosage of
essential oils indicates liquid phase applied
in the head space. The overall mean ± SE
of ACC synthase activity (nmol/g/h) of
control was 1.38±0.21.
３Effect of Essential Oils on In Vitro Activities of 1-Aminocyclopropane-1-carboxylate (ACC) Synthase and ACC Oxidase
Fig. 2 Effect of essential oils on in vitro activity
of ACC oxidase obtained from wounded
winter squash mesocarp tissues. Dosage of
essential oils indicates liquid phase applied
in the head space. The overall mean ± SE
of ACC oxidase activity (nmol/g/h) of
control was 5.11±0.69.
４ A. B. M. Golam RABBANY, Fusao MIZUTANI and Sojiro CHIKAIZUMI
It is well known that plants emit carbonyl compounds
including pentanal, hexanal, nonanal, and decanal
(Kesselmeier and Staudt, 1999). Matsui and Hatanaka
(2005) reported biological activities of green leaf volatiles
in which C6 aldehydes and alcohols are main constituents.
Among them 1-hexanal is also involved. If these aliphatic
aldehydes are produced in the plant tissues by some
factors such as wounding or senescence, non-enzymatic
breakdown of ACC occurs leading to ethylene production.
The ethylene thus produced seems to trigger the
subsequent physiological cascades like autocatalytic
ethylene production.
References
Baur, A. and S. F. Yang (1969) Ethylene production from
propanal. Plant Physiol. 44: 189－192.
Beauchamp, C. and I. Fridovich (1970) Mechanism for the
production of ethylene from methional. J. Biol. Chem. 245:
4641－4646.
Boller, T., R. C. Herner and H. Kende (1979) Assay for and
enzymatic formation of an ethylene precursor, 1-
aminocyclopropane-1-carboxylic acid. Planta 145: 293－
303.
Corteau, R. (1987) Biosynthesis and catabolism of monoterpe-
noids. Chem. Rev. 87: 929－954.
Hyodo, H., K. Tanaka and K. Watanabe (1983) Wound-
induced ethylene production and 1-aminocyclopropane-1-
carboxylic acid synthase in mesocarp tissue of winter
squash fruit. Plant Cell Physiol. 24: 963－969.
Jerie, P. H. and D. J. Chalmers (1976) Some characteristics of
ethylene production in peach (Prunus persica L.) seeds.
Planta 132: 13－17.
Kesselmeier, J. and M. Staudt (1999) Biogenic volatile organic
compound (VOC): An overview on emission, physiology
and ecology. J. Atmos. Chem. 33: 23－88.
Kacperska, A., and M. Kubacka-Zebalska (1993) Wound
ethylene synthesis in the stress-affected cells. In: Pech, J.
C., A. Latché and C. Balagué (eds) Cellular and Molecular
Aspects of the Plant Hormone Ethylene, Kluwer Academic
Press, Dordrecht, pp. 211－216.
Lizada, M. C. C. and S. F. Yang (1979) A simple and sensi-
tive assay for 1-aminocyclopropane-1-carboxylic acid. Anal.
Biochem. 100: 140－145.
Matsui, K. and A. Hatanaka (2005) Biological activities of
green leaf volatiles. Reg. Plant Growth Develop. 40: 52－
61.
McRae, D. G., J. A. Coker, R. L. Legge and J. E. Thompson
(1983) Bicarbonate/CO2-facilitated conversion of 1-
aminocyclopropane-1-carboxylic acid to ethylene in model
systems and intact tissues. Plant Physiol. 73: 784－790.
Rabbany, A. B. M. G. and F. Mizutani (1996) Effect of
essential oils on ethylene production and ACC content in
apple fruit and peach seed tissues. J. Japan. Soc. Hort. Sci.
65: 7－13.
Tigeny, D. T., D. P. Turner and J. A. Weber (1991) Factors
controlling the emission of monoterpenes and other volatile
organic compounds. In: Sharkey, T. D., E. A. Holland and
H. A. Mooney (eds) Trace Gas Emission by Plants.
Academic Press, Inc. San Diego, California, pp. 93－119.
Yang, S. F. and N. E. Hoffman (1984) Ethylene biosynthesis
and its regulation in higher plants. Ann. Rev. Plant Physiol.
35: 155－189.
Yosshii, H. and H. Imaseki (1981) Biosynthesis of auxin
induced ethylene. Effect of indole-3-acetic acid, benzylade-
nine and abscisic acid on endogenous levels of 1-amino-
cyclopropane-1-carboxylic acid (ACC) and ACC synthase.
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摘 要
A. B. M. ゴーラム ラバニー・水谷房雄・近泉惣次郎：
カボチャから得た ACC 合成酵素と酸化酵素の試験管内で
の活性に及ぼす精油の影響
カボチャの果肉から調製した粗酵素液を用いて，試験
管内で精油が ACC 合成酵素と酸化酵素活性に及ぼす効果
を調査した．閉じた試験管内のヘッドスペースに気化し
た精油を含む状態で酵素活性を測定した．テストした全
ての精油が ACC 合成酵素活性を抑制した．リモネンとリ
ナロールではそれぞれ２８，２５μl/l の処理濃度で５０％の活性
抑制が見られた．アルデヒド型の精油ではさらに強い抑
制活性が見られた．処理をした濃度範囲で（３．６～３６
μl/l）シトラール，１‐オクタナール，１‐ノナナール，１‐デ
Fig. 3 Stimulation of non-enzymatic conversion of ACC to
ethylene by aliphatic aldehyde 1-nonanal. Dosage of
essential oils indicates liquid phase applied in the
head space.
５Effect of Essential Oils on In Vitro Activities of 1-Aminocyclopropane-1-carboxylate (ACC) Synthase and ACC Oxidase
カナールでは５０～７８％の活性抑制が，シトロネラールで
は７８～９３％の活性抑制が見られた．一方，精油は ACC 酸
化酵素活性には抑制効果を示さなかった．しかしなが
ら，１‐オクタナール，１‐ノナナール，１‐デカナールのよ
うな脂肪族アルデヒド型の精油は非酵素的に ACC を分解
し，エチレンを生成した．
６ A. B. M. Golam RABBANY, Fusao MIZUTANI and Sojiro CHIKAIZUMI


Effect of essential oils on in vitro activities of 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase from winter squash mesocarp

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