Ehime University Institutional Repository

梶原, 克彦

Japanese

国民共同体の境界 : 第一次世界大戦の経験・総力戦のなかの捕虜

岡本, 隆

熊谷, 太郎

曽我, 亘由

就職に係る大学生の能力についての企業と大学生の認識差

和田, 寿博

中小企業振興基本条例制定と中小企業振興の課題

富田, 英司

白松, 賢

池野, 修

隅田, 学

向, 平和

鴛原, 進

ルイジアナ大学モンロー校と愛媛大学の相互国際交流を通した教員養成の実践

高橋, 弘臣

南宋臨安の食糧問題

田口, 瑞季

胡, 柏

食品残渣を主とした堆肥化の取組と環境保全型農業との連携

Ehime University (愛媛大学)博士(農学)thesi

伊藤, 慎

ITO, Makoto

（第３号様式）(Form No. 3) 
学  位  論  文  要  旨  
Dissertation Abstract 
 
 
氏名：  
Name 
伊藤 慎  
 
学位論文題目：  
Title of Dissertation 
Tabtoxinine-β-lactamによる細胞死誘導機構の解明  
 
 
学位論文要旨：  
Dissertation Abstract 
 
タバコ野火病菌 Pseudomonas syringae pv. tabaci（Pta）は非選択的毒素である
tabtoxinine-β-lactam（TβL）を分泌し、宿主植物である Nicotiana benthamiana の葉
に壊死病斑を形成する。TβL は植物のグルタミン合成酵素に作用し、細胞内に過剰
なアンモニアを蓄積させることにより細胞死を誘導するが、その細胞死誘導機構
は不明であった。しかしながら、近年、Pta が誘導する壊死病斑形成は遺伝的にプ
ログラムされた自発的な細胞死（Programmed cell death; PCD）であることを示唆す
る報告が成された。本研究では TβL による細胞死誘導機構の解明を目的とし、TβL
処理が PCD を誘導することを明らかにするとともに、ウイルス誘導型遺伝子サイ
レンシング法を用いた逆遺伝学的な解析により、N. benthamiana において TβL 誘導
細胞死に関与する植物因子の探索を行った。  
 TβL 処理が PCD を誘導することを明らかにするため、N. benthamiana に TβL を
処理し、植物における典型的な PCD である過敏感細胞死（Hypersensitive response; 
HR）のマーカー遺伝子である Hin1 の発現解析を行った。その結果、TβL 処理によ
り顕著な Hin1 の発現上昇が認められた。したがって、TβL は PCD を誘導すること
が示唆された。  
分子シャペロン Heat Shock Protein 90（HSP90）とそのコシャペロンとして機能
する suppressor of G2 allele of skp1（SGT1）と Required Mla-12 Resistance（RAR1）
は、植物‐病原体相互作用における PCD の典型的な例である HR の誘導に広く関
与することが報告されている。また、分子シャペロンである Heat Shock Protein  70
（HSP70）も病原体に対する HR に関与することが明らかとなっている。そこで、
TβL 誘導細胞死への分子シャペロン HSP70 と HSP90 とそのコシャペロン SGT1 と
RAR1 の関与を明らかにするため、ウイルス誘導型遺伝子サイレンシング法により
HSP70、HSP90、SGT1 および RAR1 をそれぞれサイレンシングした N. benthamiana
を作出し、TβL 処理を行った。その結果、HSP70 サイレンシング植物においての
み壊死斑形成の顕著な抑制が認められた。また、細胞死の指標となるイオン漏出
量の増大は、HSP70 サイレンシング植物の TβL 処理部位においては顕著に抑制さ
れた。以上の結果から、TβL 誘導細胞死に HSP70 が関連する植物細胞内情報伝達
経路が関与することが示唆された。  
PCD 誘導に関与することが報告されている植物ホルモンであるサリチル酸、ジ
ャスモン酸およびエチレンとシグナル伝達に関わる活性酸素種、一酸化窒素およ
（第３号様式）(Form No. 3) 
び mitogen-activated protein kinase（MAPK）カスケードの TβL 誘導細胞死への関与
を解析したところ、いずれの因子においても TβL 誘導細胞死への関与は認められ
なかった。  
上記の結果より、近年の植物における PCD 研究の成果は TβL 誘導細胞死に適用
できないものと考えられた。そこで、TβL による細胞死がアンモニア毒性によるも
のであるという点に着目した。動物細胞では、アンモニア処理により cAMP を介
した PCD が誘導されることが報告されている。また、様々な動物細胞において
cAMP は PCD 誘導に関与することが報告されている。そこで、TβL 誘導細胞死へ
の cAMP の関与を検討するために、cAMP 合成酵素であるアデニル酸シクラーゼを
コードする遺伝子 N. benthamiana Adenylyl cyclase（NbAC）をサイレンシングした
植物を作製し、TβL を処理した。その結果、対照植物と比較して NbAC サイレンシ
ング植物では壊死斑の形成、イオン漏出量の増大および Hin1 の発現上昇が顕著に
抑制された。以上の結果より、TβL 誘導細胞死に NbAC が関与することが明らか
となった。このことから、NbAC により合成される cAMP の TβL 誘導細胞死への
関与が示唆された。  
本研究により、TβL による細胞死誘導・壊死病斑形成は植物自身の因子である
HSP70 と NbAC が関与する PCD であることを示した。一方、TβL による細胞死誘
導は植物ホルモン等の特徴づけが進んでいる細胞死誘導機構とは異なる特性を有
していると推察された。今後、TβL による細胞死誘導機構の解析により、新奇の細
胞死誘導経路関連因子の同定を可能にすると考えられた。  


Tabtoxinine-s-lactamによる細胞死誘導機構の解明

Seriba, KONARE

(Style5)
Extended Summary in Lieu of  Dissertat ion
S e r i b a  K O N A R E
  
T i t l e  o f  D i s se r t a t ion :   
Fuelwood sys tem in  Mal i  -  communi ty percept ion  
and impact  on  fores t  s t ructure
( )  
            Fue lwood  i s  ma in  source  o f  ene rg y fo r  man y deve lop in g  
coun t r i e s  in  the  w or ld .  I t  i s  u sed  fo r  d ive r se  pu rposes  such  a s  he a t ing
house ,  bo i l i ng wa t e r,  cook in g ,  and  s o  on .  Fo r ex ample  in  Mal i ,  one  
peop le  use 291kg/ye a r  o f  fue lwood  to  sa t i s f y the  need .  Unfo r tuna te l y  
fue lwood  ex t r ac t io n  f rom fo res t  has  l ead  to  de grad a t ion  o f  fo res t  i n  
seve ra l  pa r t s  o f  t he  wor ld .  In  o rd e r  t o  mi t iga t e  the  ne ga t ive  impac t s  o f
fue lwood  ex t r ac t ion  on  fo res t , fo re s t  manager s  have  in t roduced  seve ra l  
po l i c i e s  and  innova t ion .  The y h ave  s t a r t ed  a ffo res t a t ion  and  
r e fo res t a t ion  p ro gr ams  to  ca t ch  up  t he  amoun t  ex t r ac t ed  b y fu e lwood  
ex p lo i t a t ion .  The y  have  in t roduc ed  improved  s toves  w h ich  consumed  
l e s s  fue lwood  compare  to  t r ad i t iona l  one .  The y cons ide r  some  pa r t s  o f  
fo res t  a s  p ro tec te d  fo res t ,  where  i t i s i l l e ga l  t o  co l l ec t  fue lwood .  
Though  the re  a re  those  d i ffe ren t  a t t empts ,  fue lwood  ex t r ac t ion s t i l l  
deg raded  fo res t  i n  deve lop ing  wor ld .
            In  t h i s  r e sea rch  w e  ana l yz ed percep t ion  o f  ru ra l  communi t i e s  
and assessed the  e ffec t  o f  fue lwood  ex t r ac t ion  on fo res t  s t ruc tu re ,  i n  
o rde r  to  so lve the  i s sues  happened  in  f ue lwood  s ys t em in  Mal i .  
             Our  r esea rch  s i t e s  we re  Was soro la  and  Farako ,  two  v i l l a ges  in
Mal i .  In  J anua r y 2010 ,  a  su rve y o f  10 0  peop le  was  ca r r i e d  ou t  t o  a s ses s  
v i l l age r  s op in ions  on  the  fue lwood  i s sue  and  the  l i f e s t yl e s  o f  t he  
v i l l age r s  o f  W assoro la .  In t e rv iews  were  conduc ted b y f ac e - to - fa ce .
P r inc ipa l  componen t  ana l ys es  (PCA)  o f  the  da ta  wer e app l i ed  to  an a l yz e
the  va r i a t ion  and  s t ruc tu re  o f  the  da ta .  
              In  o rde r  to  ana l yses  the  e ff e c t  o f  fue lwoo d  ex t rac t ion  on  
fo res t  s t ruc tu res ,  we  se t  r e s ea r ch  p lo t  bo th  in  p ro tec t ed  and  non  
p ro tec ted  fo res t in  Oc tobe r  2011 .  And  we  r andoml y se t  u p  5  p lo t s  o f  20
m X 20  m in  bo t h t ypes  o f  fo res t s .  To  quan t i f y s t and  s t ruc tu re ,  w e
measured  D BH (s t e m diamete r  a t  b re a s t he igh t )  o f  a l l  i nd iv idua l s  l a rge r  
than  4cm of  DBH.  W e a l so  coun ted  the  number  o f  i nd iv idua l s wi th l e s s  
1 
 
学位論文要約
Dissertation Summary
(Style5)
than  4  cm of  D BH and  number  o f  s t e ms  fo r  a l l  t he  sh ru bs  wi th in  p lo t s .
W e iden t i f i ed  a l l  t r ee s  and  sh rubs  in  the  p lo t .
               Fami l y un i t s  i n  Wasso ro l a cons i s t  o f  l a rge f ami l i e s  w i th  an  
ave ra ge  o f  6 .7  a du l t s  and  6 .5  ch i ld ren .  The  educa t ion  l eve l  o f  
i n t e rv i ew ees  i s  l ow and  84% of  the m never  a t t ended  schoo l .  Of  the  
r e sponden t s ,  83% was aware  o f  the  th rea t  t o  the i r  fo re s t  r e sourc e  and  
12% though t the re  i s  no  so lu t ion  to  the  p rob lem.  
Resu l t  o f PCA ana lys i s showed educa t ion  and  capa c i t y  t o  wr i t e  and  read  
have  h ighes t  r e l a t ed  fac to r . La rge  fami l i e s  t end  to  t h ink  the i r  
sou r round ing  fo re s t  dep le t ion  i s  d ue  to  na tu ra l  r ea sons ,  o r  sma l l  
f ami l i e s  t h ink  i t  i s  due  to  human  ac t iv i t i e s . Thi s  phenomen can  be  
ex p la ined  b y the  t r ad i t iona l  wa ys  o f  b e l i eve .
Non  p ro tec t ed  fo r es t  had  2 ,018  ind iv idua l s ( t r ees  an d  sh rubs )  pe r  
hec t a re ,  w i th  35  spec i es  and  17  fami l i e s .  There  was  n o  t r ee  wi th  a  
d i amete r  a t  b re as t  he igh t  (D BH)  l a rge r  t han  60  cm.  The  b asa l  a re a  i n  t he  
non  p ro tec t ed  fo res t i s  6 .79  m 2 .  P ro te c t ed  fo res t had 3 ,3 69  indiv idua l s
pe r  hec t a re ,  w i th  5 1  spec i es  and  19  f ami l i e s .  I t s  l a rges t  t r ee  had  a  D BH 
of  165  cm.  The  bas a l  a rea  in  the  p ro te c ted  fo res t was  16 . 15  m 2 /ha .
These  r esu l t  r eve a l tha t  p ro tec t ed  fo res t  had  mor e t r e e s ,  b igge r  t r e es  
and  h ighe r spec ie s  d ive r s i t y  compar e d to  non  p ro tec t ed  fo res t .  We ma y 
conc luded  tha t  p ro tec t ed  fo res t  pe r fo r m we l l  t hese func t i ons .
Th is  r e sea rch  i s  p roved  tha t loca l  peop le  a re  aw are  abou t  h igh
fue lwood consumpt ion and  tha t  p ro tec t ed  fo res t i s  e f f ec t i ve  in  fue lwood  
co l l ec t ion  a rea . Th i s  document i s  impor t an t  fo r  po l i c ymak er s .
2 
 


English

Fuelwood system in Mali : community perception and impact on forest structure

Khaoian, Peerapon

（様式５）(Style5) 
 
学位論文全文に代わる要約 
Extended Summary in Lieu of Dissertation 
 
氏名： 
Name 
 
Khaoian Peerapon 
 
学位論文題目：    
Title of Dissertation 
DEVELOPMENT OF LOW-FISH MEAL PRACTICAL DIET FOR 
YELLOWTAIL 
(ブリ用低魚粉実用飼料の開発) 
 
 
 
 
Yellowtail Seriola quinqueradiata is one of the economically important aquaculture fish species in 
Japan. Its carnivorous nature demands a high protein diet, which typically contains high concentration of fish 
meal (FM). Recently, high demand and limited supply for FM resulted in inflating FM cost. Therefore it is 
necessary to develop low-FM diet with inexpensive ingredients such as conventional plant proteins. In 
previous studies, application of low-FM with plant origin feed ingredients has never been succeeded. Later, it 
has been reported that a non- or low-FM soy protein concentrate (alcohol purified defatted soybean meal) 
based diet produced good growth performance and physiological conditions in yellowtail when taurine was 
supplemented in the diet. Thus, it is inferred that the reasons of unsuccessful results of low-FM with plant 
origin feed ingredients diet might be a taurine deficiency, whereas this hypothesis has never been tested. 
In the present study, a series of experiments were conducted with the aims: to examine if low-FM diet 
with practical plant origin ingredient namely soybean meal (SBM) and corn gluten meal (CGM) supplemented 
with taurine maintains normal growth and physiological conditions of juvenile and one-year-old yellowtail, 
respectively.  
 
Feeding soybean meal diet supplemented with taurine to yellowtail affects growth performance 
and lipid digestion  
 Feeding experiment was conducted to determine the effects of soybean meal with supplementation 
of taurine on growth performance and lipid digestion. Three diets (FM, SBM, and SBM + taurine;T) were fed 
to yellowtail (42 g). The mean body weight and specific growth ratio were significantly lower in SBM+T-fed 
fish compared with FM-fed fish at the end of experiment. Taurine supplementation of SBM improved lipid 
digestion and absorption, but did not improve growth performance.  
These results indicate that apart from lack of taurine, SBM contains other factors responsible for 
inferior growth, and impaired lipid metabolism was not a major factor in the reduced growth of yellowtail fed 
SBM. Replacement of FM with SBM-based diet must be limited in appropriate proportion, even though taurine 
was supplemented in the diets and meet to require for yellowtail. 
 
 
 
 
（様式５）(Style5) 
 
Effects of low-fish meal practical diet on growth and physiological condition of young yellowtail 
The effects of taurine supplementation and palm oil replacement in low-FM diet for yellowtail were 
examined in a feeding experiment using four different diets. The control diet had an FM content of 600 g/kg 
(FM60). The low-FM diets contained 350 g/kg FM, 170 g/kg corn gluten meal, and 170 g/kg defatted soybean 
meal (FM35). Taurine was supplemented in FM35 at 7.5 g/kg (FM35T). Fish oil in the FM35T diet was 
replaced with palm oil (FM35TPO). Juvenile fish (initial body weight 236 g) were fed these diets for 36 weeks. 
The mean body weight and specific growth rate were not significantly different among the treatments until 24 
weeks. However, they were significantly higher in FM35T-fed fish compared with FM35-fed fish at week 36. 
The feed intake levels of FM35- and FM35T-fed fish were comparable or higher than that of FM60-fed fish 
whereas the intake levels of FM35TPO-fed fish were always significantly higher than those of FM60- and 
FM35T-fed fish (P < 0.05). The tissue taurine concentrations of FM35-fed fish were significantly lower than 
those of FM60-, FM35T-, or FM35TPO-fed fish in all tissues examined at all sampling times (P < 0.05). 
FM35TPO-fed fish were leaner than FM35T-fed fish, as evident from the lower lipid concentration in the 
muscles and whole body. The lower feed conversion ratio of FM35TPO-fed fish compared with FM35T-fed 
fish suggested that the lower lipid absorption from the FM35TPO diet was due to the inclusion of palm oil.  
These results suggested that taurine supplementation is necessary during long-term feeding of 
yellowtail with low-FM diet. Despite the economic advantage of palm oil compared with fish oil, use of palm 
oil should be limited because it has lower digestibility in cold-water conditions.  
   
Effects of low-fish meal diet supplemented with taurine on growth and taste of one-year-old 
yellowtail  
 Three diets (FM60, FM35, and FM35T) were fed on one-year-old yellowtail (2.9kg) for 18 weeks 
during winter low water temperature season. Fish fed FM35 could maintain comparable growth and 
physiological conditions to fish fed FM60. There are no significantly differences in hepatosomatic index (HSI), 
gallbladder somatic index (GBSI), viscera somatic index (VSI) and biliary bile salt concentrations among the 
treatment at weeks 7 and 18. The taurine concentration in the liver of FM35-fed fish was significantly lower 
than that of FM60- and FM35T-fed fish at week 18, suggesting that the dietary taurine concentration of FM35 
diet was not enough to maintain the tissue taurine concentration. The taste test was performed with or without 
soy sauce in order to clarify the taste differences between fish. The taste of dorsal part was no significantly 
different among the treatments, whereas the taste of ventral part of FM35-fed fish had significantly different 
compared with FM60- and FM35T-fed fish.  
The present study could be summarized that dietary taurine supplementation may not be necessary to 
maintain growth and physiological conditions of the one-year yellowtail at least for 18 weeks in winter season. 
The taste of FM35T-fed fish was better than FM-35 fed fish and comparable to FM60-fed fish. 


DEVELOPMENT OF LOW-FISH MEAL PRACTICAL DIET FOR YELLOWTAIL

Ehime University (愛媛大学)博士(学術)thesi

ピヤーパット, トライソンティ

PIYAPAT, Trisonthi

（第３号様式）(Form No. 3) 
学  位  論  文  要  旨  
Dissertation Abstract 
 
 
氏名：  
Name 
Trisonthi Piyapat 
学位論文題目：  
Title of Dissertation 
Studies on Bioactive Compounds Found in Litsea cubeba Fruits
(Litsea cubeba 果実に含まれる生理機能性物質に関する研究 ) 
 
 
学位論文要旨：  
Dissertation Abstract 
 
Litsea cubeba (Lauraceae) is a plant distributed in Himalaya, southern China, 
Taiwan, Japan and South East Asia. All part of this plant has been used as traditional 
medicines. L. cubeba is also well known as an essential oil source. The fruit essential 
oil, May Chang oil, is an important aromatherapy component and a raw material for 
citral, ionones and vitamin A production. In this study, we determined antioxidant 
activities of L. cubeba fruit essential oil components, anticancer activity of methanol 
extract of L. cubeba fruits residue from essential oil extraction. Finally, anticancer 
bioactive compounds were isolated and analyzed by spectroscopic methods. 
For the antioxidant activity, the essential oil components which exhibited radical 
scavenging activity (DPPH assay) are geranial, neral, geraniol, (-)-pinene, 
-citronellol, -pinene, citronellal, linalool, ocimene and -terpinene. The highest 
activities were exhibited by ocimene and -terpinene. For the activity to prevent lipid 
oxidation (-carotene bleaching assay), most of the compounds exhibited the activity 
but geranial neral geraniol nerol (-)-pinene -caryophyllene and -terpinene were 
significantly more effective than other components. 
The essential oil extracted from L. cubeba fruits was found to be less effective 
than the fruit residue methanol extract in cytotoxicity against HeLa cells. Therefore, 
the methanol extract was then fractionated by AmberliteTM XAD-7 column 
chromatography (mobile phase: 20-100% methanol). Only four fractions eluted by 
80-100% methanol (4B, 5A, 5B and 5C) were effective against viability of HeLa cells 
and they also exhibited activity to promote G1-S phase cell cycle arrest (BrdU assay). 
Late apoptosis was observed according to leakage of LDH in cell population treated 
with these samples. Finally, these fractions were proven to be effective in induction of 
effector caspase-3/-7 activation in HeLa cells. 
However, the effective fractions isolated by using XAD-7 resin as stationary 
phase were highly complex. So, fractionation of the methanol extract was repeated by 
using silica gel (45~75μm particle size) as stationary phase instead. Elution was done 
（第３号様式）(Form No. 3) 
by using mobile phase gradient of hexane and diethylether to give 20 fractions. 
Fraction 11 and 12 eluted by 50% diethylether were cytotoxic against HeLa cells. The 
major cytotoxic compound in these fractions [(+)-6-(4-hydroxy-4-methyl-2-pentenoyl)- 
4,6-dimethyl-5-(3-methyl-2-butenyl)-1,3-cyclohexadienecarbaldehyde, (Compound 1)] 
was successfully isolated by preparative HPLC and identified for chemical structure by 
spectroscopic methods (1-D, 2-D NMR, IR, UV, HR-ESI-MS). This compound structure 
has not been reported elsewhere, and it was named as cubelin (C20H28O3). This 
compound exhibited activity against HeLa cell viability and proliferation (IC50: 34.43, 
30.49 M). The changes in nuclear morphology which indicates apoptosis were clearly 
observed. The presence of cleaved caspase-3/-7, caspase-8 and caspase-9 in the cubelin 
treated population indicated the potential of the compound to induce apoptosis in HeLa 
cells via both intrinsic and extrinsic pathways. 
 
 Other than cubelin, there are several other cytotoxic compounds isolated from 
methanol extract of L. cubeba fruits that are not yet successfully identified. Firstly, 
compound 2, 3 and 4 from the same fractions as cubelin exhibited cytotoxicity with 
IC50 of 41.91, 52.50 and 60.18 g/mL. Another two compounds (5 and 6) were obtained 
by washing the column, after elution of hexane/diethylether gradient, with 10% acetic 
acid in ethanol. The compounds were isolated by using preparative HPLC. Compound 5 
and 6 exhibited activity against HeLa cell viability (IC50: 1.58, 2.25 g/mL) and cell 
proliferation (IC50: 0.77, 0.89 g/mL). They also exhibited caspase-8, -9 and -3/-7 
inducing activity at the similar level as camptothecin. However, because of very low 
yield percentage of 5 and 6, the obtained amount is not possible for structure analysis 
by NMR spectroscopy. 


Litsea cubeba果実に含まれる生理機能性物質に関する研究

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