全 文 ：第 28卷 第 1期
2007年 1月 　　　　　　　　　　
华南农业大学学报
Journa l o f South China Agricultural University
　　 　　　　　　　　Vo.l 28, No. 1
Jan. 2007
　R eceived date:2005-11-12
Biograph ies:A ISHA H abita(1978— ), m a le, m aster;Correspond ing au tho r:ZENG L ing(1949— ), fem a le, p rofe ssor, E-m ail:
zeng ling@ scau. edu. cn
Foundation item:Natural Sc ience Foundation of Ch ina (38770516 and 39930120)
Deterrent Effect of EssentialO il Extracted from
Dolichandrone cauda-felina onOviposition of
L iriomyza sa tivae B lanchard (D iptera:Agromyzidae)
A ISHA H abita, LU Yong-yue, ZENG Ling
(Lab of Insect Eco logy, South China Ag ric. Un iv. , Guangzhou 510642, China)
Abstrac t:Ov iposition-repe lling activ ity o f e ssentia l oils ofDolichandrone cauda-felina (H ance) Ben th. ＆ H ook. F. , be-
havioral tests u sing four-arm ed o lfactom e te r and e lecroantennog ram s(EAG) response s using EAG reco rding technique in L ir-
iomyza sa tivae, w ere a ll studied. The bioassay resu lts show ed that the repe llent effec t o f the vo la tile wa s dose-dependent;the
best effe ct was at do sage of 5μL, IIPC and repe llen t ra tew ere 0. 289, 55.14% respective ly. O lfactory re sponse o f L. sati-
vae ind ica ted the strong repe llen t effec t of the essentia l o il, and no patte rns differences be tw een m ale and fem a le. The EAG
reco rding produced ana logous results:the essential oil e lic ited an EAG response in leafm ine rs adu lts, and showed a do se de-
penden.t GC M/ S ana lysis o f e ssentia l o il indica ted that there w ere 21 known com pounds;the d ibu ty l phtha late had the high-
e st re lative concentration (56. 55%).
K ey words:L iriom yza sativae;Dolichandrone cauda-felina;ov iposition;deterren t effec t
CLC num ber:S482. 3　　　　　Docum ent code:A　　　　A rtic le ID:1001-411X (2007)01-0058-05
猫尾木挥发油对美洲斑潜蝇产卵驱避作用研究
A ISHA H abita, 陆永跃 , 曾　玲
(华南农业大学 昆虫生态研究室 ,广东 广州 510642)
摘要:采用室内生物测定和电生理学的方法研究了猫尾木挥发油对美洲斑潜蝇产卵驱避作用以及美洲斑潜蝇对该
挥发油的行为反应和触角电位活性.生物测定结果表明 , 每棵豆苗施用 1 ～ 5μL挥发油 , 大部分处理的落卵量明显低
于对照 , 其中 5 μL剂量效果最好 ,干扰作用指数( IIPC)和驱避率分别为 0. 289、55.14%. 这表明挥发油对美洲斑潜蝇
具有明显的产卵驱避作用.四臂嗅觉仪测试中对挥发油的第一选择率明显较低 , 处理的停留时间大部分低于对照 ,
进入次数仅在 250 mL /m in流量时处理明显降低 .雌雄虫对挥发油的嗅觉反应无差异. 美洲斑潜蝇对挥发油表现明显
的触角电位活性 ,且随着挥发油浓度增大而增大. 触角电位活性与挥发油量之间的关系符合模型Y =43. 414e0. 369 1X .
应用 GC-M S分离出猫尾木挥发油主要含有 21种化合物 , 其中邻苯二甲酸二丁酯含量最高 ,占 56.55%,其次为邻苯
二甲酸二乙基己酯和 1 -壬烯 - 3 -醇 ,分别占 9. 8%和 6.64%.
关键词:美洲斑潜蝇;猫尾木;挥发油;产卵驱避
　　P lants con tain a large rese rvoir o f chem ica l struc-
tures w ith b io log ical ac tiv ity. Researches w ith alterna-
tive methods fo r contro lling insect pests and especially
food a ttractants, oviposition dete rrents, m ating disrup-
tion using sex pheromones and antifeedants have p ro-
gressed g rea tly in recent years
[ 1]
. A ccording to Akh tar
and Ism an
[ 2] , plan t secondary compounds have been
the sub ject of tho rough investiga tion for the past 30
years in an effort to d iscover new sources o f bo tan ica l
insecticides and antifeedants. W ell-documented reco rds
show that before 1850, 20 plant species be long ing to 16
diffe rent fam ilies w ere used to control the ag ricu ltura l
and horticu ltura l pests in Western Eu rope and Ch i-
na
[ 3-4]
. A ctua lly the use o f plan ts compounds is still un-
der deve lopment, and only time w ill te llwhe ther or no t
they w ill w idely adopted
[ 5]
. Only mo re than 250 000
plant spec ies on ou r p lane t have been p roperly evalua-
ted fo r use in pest con trol
[ 6]
. The rich know ledge of
plants w ith pesticide properties w as no t lo st in Ch ina,
as evidenced by a recent repo rt stating that in Ch ina,
diffe rent parts o r extrac ts of 276 p lant species are used
as pe sticides
[ 7]
. Some insectic ides of plan t o rig in w ere
used on a large sca le befo re they w ere out competed by
synthe tic insec ticides;nico tine, rotenone and py rethrins
have been extensive ly used and w ere effective insecti-
cides because they degrade rapid ly, do no t accumu late
in the food chain
[ 8]
. In the present study the repellen t
activ ity of essen tial oils fromDolichandrone cauda-felina
againstLiriomyza sa tivae (B lancha rd)was investiga ted.
The olfactory and e lectrophysiolog ical response w as also
dete rm ined, and the chem ical composition of the essen-
tial o ils w as de te rm ined by GC M/ S ana lysis.
1　Mater ials and m ethods
1. 1　Insect collection
Infested leave sw ith L. sa tivae larvae w ere co llect-
ed from the coun tryside in Guangzhou. A fter larvae
w ere changed into them , pupae w ere co llected in sma ll
tubes, and new emerged adu ltsw e re ready to te sts.
1. 2　Extracting method of essential oils
Fresh leave s o f D. cauda-felina were co llected
from the campus of South ChinaAg ricultu ra lUn iversity,
and cut to sma ll pieces then steam d istilled. The m ix-
ture o f o il and w ater obtained w as separa ted using e thy l
ethe r, then dried by anhyd rous N a2 SO4 and the so lven t
w as removed using evapo rator ro tator. The crude of es-
sentia l o ils w as sto red in a refrigerato r at 3-4 ℃ until
used.
1. 3　B ioassaymethods
K idney bean plan tsPhaseolusvagaris L. w ere used
as tested host plan ts, w ith tw o seedlings in a cup each
seedling has two leaves. A known quantity of essentia l
oils w as app lied a t a filte r pape r beside the seedlings
placed in a bioassay cage, adu lt leafm iners w e re intro-
duced in the cage, and 10% (φ) o f honey w as prov id-
ed as a supplemen t food to the adults. A fter 24 h num-
ber of eggsw as checked under the m icro scope. The ex-
perimen tw as conducted a t(27±1) ℃.
1. 4　Behavioral test
Four-armed olfactometer w as used to test the be-
hav ior o f the bo th sex o f the adult leafm ine r, one arm
w as se lected as the trea ted arm , and the othe r 3 a rms
w ere untreated. The hum idified air stream , d ra ined
th rough a ho le in the center o f device, was se t a t 60,
100, 200 and 250 mL /m in successfully for each arm ,
in each test flow , 20 adu ltsw ere ind iv idua lly introduced
in to the chamber through the exit hole and test timew as
10m in fo r every adult;the adu lt w as moved freely in
the chamber, the time spen t in each arm , the numbe r
of entries and the first cho ice of the test adultw e re re-
co rded th rough the softw are OLFA.
1. 5　E lec trophysiologica l record ing
The elecroantennograms (EAG) technique and the
odo r de livery system are as describ ing by Zhao and
K ang
[ 9]
. The tested insects head w as removed, and
the tip of the a rista cu t o ff, the reference electrode w as
positioned into the haemocoe l of the cranial cav ity,
while the recording e lec trode w as connected w ith the cu t
arista. A constant flow (0. 1 L /m in) of charcoa l-fil-
tered and hum idified compre ssed air w as passed ove r
the antenna through a disposable nozzle positioned 1 cm
from the antenna. Test substances we re in troduced into
the air stream by inserting the tubing o f the ca rtridge in-
to the hole of delive ry tube and in jecting the odo r stimu-
lation w as 0. 2 s, the signalw as amplified 10×by a u-
nive rsa l AC /DC Un-06 and v iew ed in the compu te r
screen. EAGsw ere recorded w ith a Syntech EAG car-
tridge reco rd ing sy stem for w indow s. Differen t concen-
trations w ere prepared by dilution o fD. cauda-felina es-
sential oils w ith paraffin o il;they w ere applied to filter
pape r, which placed inside Pasteur pipe ttes ( l =
15 cm), a standard stimu lus cis-3-hexen-1-ol w as test-
ed. Fo r each stimu lus EAG s w ere recorded from 10 an-
tennae of the leafm iner flies, the negative def lection val-
ues we re recorded in the computer. These values were
conve rted by measuring the pe rcentage of the response to
59　第 1期 　　　　　AISHA H abita等:猫尾木挥发油对美洲斑潜蝇产卵驱避作用研究 　　　
the accompany ing cis-3-hexen 1-ol standard.
2　Results
2. 1 　 Oviposition deterrent of D. cauda-felina
againstL. sativae adults
　　The da ta in the tab le 1 showed tha t all dosages
(except dosage 3 μL ) had a significant repe llency
effect on the leafm iners adults, and the best e ffec tsw ere
sim ilar in dosage of 1, 2 , 4 and 5 μL. The repe llency
ra tew as 57. 28%, 77. 38%, 65. 32% and 66. 83% re-
spective ly.
Tab. 1　The e ffect of ov iposition de terren t of p lan t vo latile
on adu lt leafm iners L. sativae1)
dosage /μL m ean num ber o f eggs repe llency ra te /%
1 8. 50±0. 80b 57. 28
2 4. 50±0. 50b 77. 38
3 13. 00±5. 45ab 34. 67
4 6. 91±2. 61b 65. 32
5 6. 66±3. 27b 66. 83
0 19. 92±3. 19a
　1)M eans in the sam e co lum n, fo llowed by the same letter are
not significan tly diffe rent a t level 0. 05 (DMRT)
2. 2　O lfactory response of L. sa tivae in various
flow rates of p lant volatiles
　　The resu lts o f b ioassays in the o lfactometer are
shown in the table 2 and table 3. In the control, the
leafm ine rs spentmo re time than in the trea ted a rm s. The
time spentwas sign ificantly different in different air flux,
itw as the maximum at 60mL /m in, and 250mL /m in of
air current in the contro l arms w as the lowest in treated
arms. The number of entriesw as significantly different a t
250mL /m in between contro l and trea ted arms. The first
cho ice ra te was significantly d ifferent in all diffe rent air
current flux. The resu lts in the tests show ed that essen-
tial o il had obvious deterrent e ffect on the L. sativae a-
dults. O lfactory response patterns of ma le and female
leafm ine rsw ere sim ila r in the b ioassay ( table 3).
2. 3　E lectrophysiolog ical response of L. sa tivae to
plant volatiles
　　The chem ical senses of insects are characterized by
an extremely high sensitivity and are key facto rs for the
insects behavior. A variety of e lectrophy siolog ical and
behav ioral bioassays have been deve loped to use these
fea tu res fo r scientific and practica lapplica tions. EAG is
Tab. 2　O lfactory response ofL. sativae adu lts in various flow rates of p lan t volatiles1)
air current flux /
(mL m in -1)
t /m in number of entrie s first choice
CK trea tm en t CK trea tm en t CK treatment
60 2. 80±0. 23a 1. 00±0. 54b 2. 40±0. 30a 2. 15±0. 50a 0. 95±0. 10a 0. 05±0. 10b
100 2. 67±0. 25a 1. 64±0. 73a 2. 10±0. 60a 1. 70±0. 95a 0. 80±0. 15a 0. 20±0. 15b
200 2. 60±0. 27a 0. 65±0. 49b 3. 06±0. 70a 2. 15±0. 82a 0. 90±0. 13a 0. 10±0. 13b
250 2. 72±0. 15a 0. 07±0. 05b 2. 68±0. 32a 0. 40±0. 20b 0. 95±0. 10a 0. 05±0. 10b
　1)M eans in the sam e co lum n, follow ed by the sam e le tte r a re no t sign ificantly differen t at leve l 0. 05(DMRT)
Tab. 3　O lfactory responses of fem ale and m ale of leafm iner adults in var ious flow rate s of p lant volatiles1)
air current flux
/(m L m in- 1)
t /m in number of entrie s first choice
fem ale ma le fema le m a le fem a le m a le
60 1. 61±0. 36a 2. 18±0. 54a 2. 35±0. 39a 2. 20±0. 44a 0. 50±0. 00 a 0. 50±0. 00a
100 2. 03±0. 55a 2. 29±0. 57a 2. 48±1. 06a 1. 32±0. 32a 0. 50±0. 10a 0. 50±0. 10a
200 1. 81±0. 54a 1. 44±0. 35a 2. 28±0. 57a 2. 93±0. 92a 0. 50±0. 00a 0. 50±0. 00a
250 1. 42±0. 33a 1. 37±0. 32a 1. 52±0. 40a 1. 56±0. 35a 0. 50±0. 00a 0. 05±0. 00a
　1) A pa ir ofm eans in the sam e row in the sam e index, fo llow ed by the sam e lette r are no t sign ificantly different at level 0. 05(DMRT)
a kind o f techn ique for reco rding the sensory informa-
tion. It involves measu ring changes in electrical poten-
tial ac ross the w ho le antenna. It has the advan tage of
exposing vo latile components w ithin am ix tu re o f chem i-
ca ls to all the receptors on the antenna
[ 10]
. The data o f
EAG s recorded in the experimen t ind ica ted tha t the es-
sentia l o il ofD. cauda-felina excite the sensory ce lls of
olfactory sensilla, wh ich can be asmessenge r to insects
brain to regu late its behav io r and avo id oviposition o fL.
sativae. The results show ed a lso that EAG we re dose-
dependent, and the modelY =43.414e0. 369 1X construc-
ted can describe the relationsh ip between theD. cauda-
felina essentia l oil and reduction ratio of EAG response
to the standard(Fig. 1).
60 　　 华　南　农　业　大　学　学　报　　　 第 28卷　
F ig. 1　Dose-response of L. sativae toD. cauda-felina e ssen tial o il
2. 4　Chem ica l analysis ofD. cauda-fe lina essential
oil
　　The analysis o f essen tia l o il o fD. cauda-felina
using gas ch rom a tog raphy /m ass spectrome tric show ed
the m a jo rity o f com pounds a re som e o ily e ste rs; the
dibu ty l ph tha la te had the highe st re lative concentra-
tion (56.55%), fo llow ed by di-(2-e thy lhexy l)
ph tha late (9. 8%) and 1-oc ten-3-o l (6.64%)
( tab le 4).
Tab. 4　Chem ical composition ofD. cauda-felina essen tia l o il
retain tim e
/m in
com pound nam e
m olecu lar
form u la
p robab ility
relat ive
con tent /%
3. 96 3-h exen-1-ol, (Z)-(CAS) C6H 12O 30. 23 2. 24
4. 50 hexanal (CAS) C6H 12O 29. 21 2. 39
6. 75 (+ -)-5-cyyclohexyidene-3-hyd roxy-1-pen tene C11H18O 62. 05 0. 87
7. 67 1-octen-3-ol C8H 16O 48. 56 6. 64
10. 72 linalool oxide(2) C10H18O 2 48. 25 0. 49
11. 65 linalool C10H18O 63. 21 1. 67
14. 34 ben zoic acid, 2-hyd roxy m ethy l ester(CAS) C8H 8O3 44. 86 0. 43
18. 64 10, 12-otadecad iyno ic acid C18H28O 2 6. 73 0. 68
19. 69 á-dam ascenone C13H18O 78. 09 1. 41
20. 86 à-ionon e C13H20O 38. 63 0. 73
23. 31 nonadecane(CAS) C19H40 12. 44 0. 34
27. 88 hexadecane, 2, 6, 10, 14-tetram ethy l-(CAS) C20H42 7. 39 1. 18
33. 50 d ibu tyl ph thalate C16H22O 4 55. 18 56. 55
39. 15 2-h exadecen-1-ol, 3, 7, 11, 15-tetram ethyl-, [ R-[ R* , R*-(E)] ] -(CAS) C20H40O 43. 25 0. 99
47. 15 d i-(2-ethylhexy l) ph tha late C24H38O 4 46. 72 9. 80
3. 96 3-h exen-1-ol, (Z)-(CAS) C6H 12O 30. 23 2. 24
4. 26 (Z)-3-hp tene C7H 14 10. 35 2. 81
5. 39 ben zen e, m eth oxy-(CAS) C7H 8O 94. 3 2. 07
19. 96 10-(acety lm ethyl)-(+)-3-carene C13H20O 19. 6 1. 31
30. 73 1, 2-benzened icarboxylic acid b is (2-m ethy lp ropyl) ester(CAS) C16H22O 4 39. 25 1. 02
3　D iscussion
　　D. cauda-felina (Hance) be longs to the fam ily of
B ignoniaceae, which posses aw ide of inte resting chem i-
ca ls structu res and pharmaco log ica l activ itie s
[ 11]
. D.
cauda-felina is a tree w ith 5-15 m tall, young bran-
ches, leaves and pedunc les ye llow-b row n, flow ers
ye llow or ye llow red, leaves 30-55 cm , leafle ts 4-8
pairs. This species is found in sparse forests, hum id
places 900-1 200 m , d istribu ted in South and South
EastAsia. It has been used formedicine and pharma-
co logy.
In Thais trad itional med icine, the leaves and
barks a re ex terna lly used fo r treating skin d iseases as
w e ll as be ing interna lly used for ana lgesic e ffec.t In in-
vestigation done by Josh i e t al
[ 12]
for th is spec ies, they
iso lated some compounds like dehydro-alfa-lapachone,
lapachol, dehydro-iso-a lfa-lapachone, B-sitoste ro l, B-
lapachone, tecto l, paulow in and pau lw in in and pa lm i-
tone from the alcoho lic ex tract of stem-heart w ood.
Early, T ripe tch e t a.l [ 13] isola ted 19 compounds, 5 w ere
new verbacoside derivatives (markham iosides A-E);
one was a new hydroqu inone deriva tive (m arkham ioside
F) a long w ith 13 known compounds from the leaves and
branches of this p lan.t S ix classes o f compounds have
been identified:qu in ines, flavonoids, te rpenoids, iri-
doids, a ph thalate and a pheno lic ester, these com-
pounds seem to bew idespread in this fam ily
[ 11]
. U rrea-
61　第 1期 　　　　　AISHA H abita等:猫尾木挥发油对美洲斑潜蝇产卵驱避作用研究 　　　
bulla e t al
[ 14]
. ind ica ted the b io log ical activity of phe-
nolic g lycosides, which w e re iso lated from A lchornea
glandu losa and show ed an antifeedant and larval g row th
effect againstSpodoptera frugiperda.
Therew as no in terest inve stiga tion for its e ffect on
insect pests. The results o f this study confirm tha t some
occurring botanicalm aterials have de fin ite ovipositiona l
dete rrence aga inst the vege table leafm iner L. sa tivae.
Them aterial tested produced a significant reduction in
the number o f eggs deposited by the insect on the trea-
ted kidney bean leaves.
The behav ioral and electrophysiolog ical response of
the L. sativae tow ard the essential o ils of this p lant con-
firm s the prev ious results ob tained. The experimen ts
show ed tha t the essen tia l oil o fD. cauda-felina , when
app lied a t high f low ra te 250mL /m in significan tly re-
duced the time spent and the numbe r of entries in the
treatment area, and applied at concentration of
0. 1mL /mL e licit a d istinct EAG response.
The chem ica l constituen t o f the D. cauda-felina
essential oils analyzed by theGC M/ S, show ed a tota l of
tw en ty essen tia l compounds tha t the dibuty l phtha late as
a major compound w ith concentration o f 56.55%.
This study is only the starting point in the long
proce ss o f deve lopment of these chem icals as a prom is-
ing componen t o f lea fm iners managemen t prog ram and
served as the foundation formo re detailed investigations
of the e ffec ts of se lec ted ex tracts on the contro l e ffec t of
Liriomyza.
R eferences:
[ 1] 　MENSAH R K, VEMEAU S, FREROT B. De te rrence of
ov iposition o f adu ltsO strinia nubila lis H ubne r (Lep idop-
tera:Py ra lidae) by a natura l enemy food supplem ent En-
v irofeast on m a ize in F rance[ J] . Interna tiona l Journa l of
PestM anagem en t, 2000, 46(1):49-53.
[ 2] 　AKHTAR Y, ISMAN M B. Comparative g row th inhibito ry
and antifeedan t effec ts o f plan t extracts and pure a llelo-
chem ica ls on four phytophagous insec t species[ J] . Jou r-
na l o f Applied Entom o logy, 2004, 128:32-38.
[ 3] 　NEEDHAM J. S cience and civilization in China[ M ] .
Cam bridge:Cam bridge University press, 1986.
[ 4] 　 SM ITH A E, SECOY D M. P lan ts u sed fo r ag ricu ltura l
pest contro l inW este rn Europe be fo re 1850[ J] . Chem is-
try and Industry, 1981, 1:12-17.
[ 5] 　PEDIGO L P. Entom o logy and pe st m anagem ent[M ] . Io-
w a:Iow a S tate Un ive rsity, 1999.
[ 6] 　CANCELADO R E. Bo tanica l insectic ides[ M ] . M inneso-
ta:Unive rsity o fM inneso ta, 2004.
[ 7] 　YANG R Z, TANG C S. P lants used for pest contro l in
China, a lite ra ture review[ J] . Econ Bo t, 1988, 42:376-
406.
[ 8] 　 SCHOONHOVEN L M , JERMY T, VAN LOON J J A.
Insec t-plan t bio logy:F rom physio logy to evo lution[ M ] .
London:Chapman and H a ll, 1998.
[ 9] 　ZHAO Y X, KANG L. Ro le o f p lant vola tiles in host p lant
loca tion of the leafm ine r, Liriomyza sa tivae (Diptera:
Agrom izydae) [ J] . Phy sio log ical Entom o logy, 2002, 27:
103-111.
[ 10] LUSZNIAK M , P ICKETT J. Self-defense fo r p lants[M ] .
Cambridge:Royal Soc iety of Chem istry, 1998.
[ 11] RASADAH M A. S tudies in the chem istry and b io log ica l
activ ity of someM a laysian B ignon iceae[M ] . London:Uni-
versity of London, 1997.
[ 12] JOSH I K C, SINGH P, PARADASANI R T. Chem ica l
constituents o f the stem heart wood o fM arkham ia stipula ta
[ J] . P lantaM edica, 1978, 34:219-221.
[ 13] TRIPETCH K, RIJOJIK, KAZUO Y. Pheno lic g lycosides
from M arkham ia stipula ta [ J] . Phy tochem istry Jou rna l,
2002, 59:557-563.
[ 14] URREA-BULLA A, SUAREZ M M , MORENO-MURIL-
LO B. B io log ica l ac tiv ity o f pheno lic com pounds from A l-
chornea glandulosa[ J] . F ito te rapia, 2004, 75(3-4):392
-400.
【责任编辑　周志红】
62 　　 华　南　农　业　大　学　学　报　　　 第 28卷