Anti-Insect Activity of the Methanol Extracts of Fern and Gymnosperm

Available online at www.sciencedirect.comAgricultural Sciences in ChinaScienceDirect2010, 9(2): 249-256February 2010Anti-Insect Activity of the Methanol Extracts of Fern and GymnospermHUANG Su- qing', ZHANG Zhi- xiang, LI You-zhi-2, LI Yu-xial and XU Han-hong'' Key Laboratory of Natural Pesticide & Chemical Biology, Ministry of Education/South China Agricultural University, Guangzhou510642. P.R.China2 College of Bio-Safery Science and Technology, Hunan Agricultural University, Changsha 410128, P.R.ChinaAbstractWith environmental and food safety issues becoming one of the greatest concerns of the public, active plant compoundsplay more and more important roles in pest contolling. Fem are of great significance for application as new pesticides dueto their particular status in plant taxology and co-evolution with insects. In this study, methanol extracts were distilledfrom wild fern and gymnosperm of 54 species of 22 families collected from Shiwandashan Mountain in Guangxi ZhuangAutonomous Region of China. Insecticidal activities of the methanol extracts were assayed on both adult house fly(Musca domestica) and mosquito (Aedes albopicrus). Significant insecticidal activities were exhibited in the methanolextracts of five species, Cupressus funebris (leaves and stems), Cycas acuminatissima (roots), Keteleeria fortunei (leavesand stems), Onychium japonicum (whole plant), and Pinus taiwanensis var. Daming shanensis (leaves and stems).Applications of the extracts of these plants resulted in higher than 50% mortalties in 4th instar larvae of A. albopictus at24 h after treatment. The methanol extracts from 13 species possessed insecticidal activities against the adult of M.domestica at 48 h after treatment with higher than 90% mortalities. The extensive screening results showed that these femand gymnosperm were highly potential to be botanical insecticides. The findings provide a feasible and valuable basis forfurther investigation.Key words: plant extracts, anti-insect activity, Musca domestica, Aedes albopictusMelia toosendan and dried flowers of RhododendronINTRODUCTIONmolle to control rice pests in China. In recent years,agrochemical companies and researchers have beenPesticide is an important factor to maintain stable cropsearching for natural products for the development ofyield, but many pesticides are highly toxic and havenew insecticides. They expect to find botanical com-long-term persistence in environment. More thanpounds that are potential to be used as biological insec-8 x 107 t pesticides have been used around the worldticides (Secoy and Smith 1983; Xu 2001; Yang andsince the 1940s. They played an important role in cropChang 1988).protection; meanwhile they also polluted the environ-Botanicals are an important source of insecticidesment (PascualVillalobos and Robledo 1999).(Arroyo 1995). Plant secondary metabolites play an im-Plant materials have been used to control insect pestsportant role in protection of the plants from being dam-in China since ancient times. Even today, farmers stillaged by pests, germ and adverse climate. Some plantuse the dried stems of Nicotiana tabacum, leaves ofsecond中国煤化工idal, hormonal orTYHCNMHGReceived 13 April, 2009 Accped 25 November, 2009HUANG Su-qing. Postdoctor, E-mail: hsuqing@ 126.com; Correspondence XU Han-hong, Professor, Ph D. Tel: +86 20 -85285127, Fax: +86-20- 38604926, E mail:hhxu@scau. cdu.cn02010.CS. Mryts rseed Ptshebyesresereddot: 10. 151621290710090250HUANG Su-qing et al.anti-feeding activities against pests, which attract the greateach plant was extracted thrice with 250 mL methanolattention of researchers (Pascual-Villalobos and Robledoat room temperature 25- 30°C for 72 h and then filtered.1998). Many plant secondary metabolites, such as pyre-The mixed filrate was concentrated by a rotary evapo-thrin and rotenone, have been used to repel pests. Therator at 40°C to dryness. All extracts were stored atmost successful one is azadirachtin, which is isolated4°C. The yields of individual methanol extracts arefrom the seeds of the neem tree Azadirachta india (Xudescribed in Table 1.2001).Effects of botanical pesticides on pests areInsectscomprehensive, involving a wide variety of biologicalprocesses, such as feeding reduction, moltingMusca domestica Adults of Musca domestica weresuspension, death of larvae and pupae, and sterility offed with water and mixture of sugar and milk powderemerged adults (Xu 2001). In long run, plant materialsin a thermostatic chamber with a constant temperaturecan offer a better solution to reduce the damage byof 28°C. Larvae were fed with artificial diet contain-pests than synthetic pesticides. Compared with syn-ing yeast, milk powder and wheat bran and kept in athetic pesticides, plant materials usually are less haz-thermostatic chamber with a constant temperature ofardous to environment and non-target organisms, such28°C and a photoperiod of 12-h light and 12-h darkness.as mammals and the pests' natural enemies.Aedes albopictus Adults of Aedes albopictus wereTo discover efctive“green" pesticides, many plantsmaintained in 30 cmx 30 cmx 30 cm glass cages, con-of different species have been analyzed for their activi-tinuously provided with 10% sugar solution in a jar withties against pests. As compared to the synthetica cotton wick. In the 5th d after post- emergence, theinsecticides, researchers expect to find more selectiveadults were deprived of sugar for 12 h and providedand less persistent active compounds from plants. Iwith a mouse placed in resting cages overnight for fe-will be beneficial for both environment protection andmales to feed blood. Larvae were reared in plastic traysagricultural product consumers, although natural prod-containing tap water and fed with a diet of yeast untilucts are not always harmfulless. In this study, wethey pupated. The pupae were transferred from thescreened a large pool of ferm and gymnosperm plantstrays to a cup containing tap water. When the adults(54 species of 22 families) for anti-insect activities andemerged, they were placed in screen cages. All thediscovered that some species contained compounds thatrearing was done in a climate chamber at (28 +2)°Cwere highly toxic to the insects tested.with a 14:10 L/D photoperiod and relative humidity of75-85%.MATERIALS AND METHODSBioassaysPlant materialsEach of the extracts was assayed for anti-insect activ-ity following the below procedure:Plants were collected in Shiwandashan Mountain of(1) M. domestica bioassays: House fly (M. domestica)Guangxi Zhuang Autonomous Region of China duringadults (males and females) from a laboratory cultivateApril 2006. The plant materials were dried naturally,without any known insecticide resistance were used 3grounded with a mill to about 400 um powders andd after emergence. The standard assay was performedstored at room temperature in hermetically sealed plas-by placing cane sugar granules (1 g) covered with thetic bags before extraction.tested extract (0.01 g) dissolved in acetone in a 1-mLvial. Acetone alone was used as a control. When thePreparation of crude extractsacetc中国煤化工1 leaving the efece-tiveFurface of the sugarTo obtain the extracts from dried plant materials, threegrantJMYHCN MH CSduced and he vialstage percolation was performed. Powder (50 g) fromwas closed with a screw cap. Mortality in these inges-Anti-Insect Activity of the Methanol Extracts of Fern and Gymnosperm251Table 1 Insecticidal activity of the plant extracts at a dose of 1 000 mg L' against the 4th instar larvae of A. albopictusMortality')Emeragence rateYield .TreatmentFamilyPart别)(%)afier treatment (%)(剐SE北SEx+SECK0.00+0.00v98.331.67a98.31.67 aAbacopteris rubraThelypteridaceaeWhole plant0.13.33土1.67 mnopqrstu 70.00+2.89 defghj70.0012.89 ghjklAdiantum caudatumAdantaceac9.35.90土1.32 lmnopqr81.71土4.44 cdefAdiantum, flabelluatumAdantaceae.53.33土1.67 mnopqrstu 70.00+ 2.89 defghij70.00+2.89 fghjil648.33.33 hijkJmnm88.33+1.67 be8.33+1.67bAngiopteris muralisAngiopteridaceae Leaves2.212.67+6.23 ghijilm75.005.00 cdeghAngiopteridaceae Stems7.73.12出 1.57 mnopqrstu 69.642.58 efghjk61.74土3.37 ijklmmoAspenium griffihianumAspleniaceaeRoots9.123.94+6.99 efghi66.21土1.93 ghijklmno 66.21+ 1.93 ghijilmAspenium 8rifithianumLeaves8.822.78+5.47 efghi61.85+3.15 hijklmnopq 58.15+6.85 hijkimnAspenium normaleAspleniaceac0.310.92+3.72 ghjklimo 76.89+3.31 cdefgh76.89+3.31 cdeighBlenchnum orientaleBlechnaceat4.44.29+2.97 lmnopqrst 89.67 +2.36 b86.10土3.64 beBlenchrum orientaleBlechnaceaeLeaves and Stems6.90.00:0.00v .79.21土5.57 edefgh79.21 t5.57 edefghCephalotaxus monniCephalolaxaceatLeaves and stems1.23土 1.23 nopqrstuv79.73+4.88 edefg78.4014.46 cdefghuariandi8.413.48土4.36 ghijlm76.85+3.43 cdefgh76.85 +3.43 ceighiCibotium baromeizDicksoniaceaeLeaves and stems 13.288.10+1.82 be .88.10+1.82 bCibotium barometzDieksoniaceae7.928.45+8.80 cfgh36.85 士12.22 jklmnopqrst 36.85土 12.22 mnopqrCupressus funebrisCupresaceaeLeaves and stems 10.454.52+ 10.60 bed45.48士 10.60 ijimnopqrs 45.48 士10.60 klmnopqCyathea podophylaCyatheaceae0.716.73+2.42 ghijil74.76+ 1.53 cdeghi74.76+1.53 cdefghCyathea spinulosaCyatheaceat3.530.48+21.27 bcdef82.2916.65 bede82.29+6.65 bcdeStems16.20.00+0.00 v68.50土3.62 fghijklm68.50+3.62 ghjkImCycas acuminatissimaStemg1.67土 1.67 nopqrstuv 81.67 +7.26 cdef81.67+7.26 edefLeaves .,.781.13+3.06 cdef77.71土1.46 cdelgCyeas acuminarissima3.86553+1.72b12.96+2.04 rstu11.81+2.13uCyeas debaoensis3.439.26土12.38 cdef59.13+8.45 hijklmnopq 59.13+ 8.45 hjklmnoCycas revolute.78.33+4.41 hjklmno 68.33土 1.67 fghjklm 68.33+ 1.67 ghijkmoCyeas revoluteCyathceaceac20.00+2.89 fghijik66.67 士4.41 ghijkImnop 66.67 14.41 ghjklmn3.66.67+1.67 jkmnop 63.33 1.67 ghikmnop 61.67+1.67 hjkmnoCyclosorus heterocarpusThelypteridaceae Whole plant2.66.67 +3.33 ijklmnopq 83.334.41 bede80.00+2.89 cdefDicranopteris amplaGleicheniaceac30.26+2.90 efg60.13 +5.07 hijklmnopq 60.13 土5.07 hjklmpoDicranopleris dichomaGleicheniaceae11.25.09+2.89 Imnop75.95+ 1.65 cdefgb75.95 土1.65 cdegh8.51.33土 1.33 nopqrstuv 80.49x 5.00 edefg80.49+5.00 cdefgDicryocline giffthii8.614.28 +5.13 ghijkim75. 63+6.20 cdefgh72.67+3.81 defhijkDrynaria fortuneiDrynariaceae0.23.33土 1.67 mnopqrstu 70.00+2 89 defghij70.00+2.89 fhijkalEquiserum difusumEquisetaceae9.42.33土 1.26 noprsuv 74.14+6.52 cdefghi71.51 士4.00 fghijkFokienia hodginsiiCrpesaceacLeaves and strms 17.333.324.67 defg62.51 +7.52 ghijklmnop 57.18士3.62 jklmnopHicriopteris chinensis13.36.67土1.67 ijkimnop76.67+6.01 cdefgh75.00+5.00 cdefghHumata tyermanniDalliaceae9.57 +5.22 hjikimn74.37土1.89 cdefghiHymenophyllum barbarum Hymenophyllaceae15.15.00土2.89 lmnopqr76.67士3.33 cdefgh75.00土2.89 ecdefghiBarks .8.23.20.0.00v67.17+7.41 fghijklmn67.17 +7.41 ghjklmn4.760.98土13.32 bed55.93 +9.26 jklmnopgr 51.39 +8.45 jkmnopLyeopodium calavatumLycopodiaceae6.19+3.12 ijklmnopq 79.21 +6.00 cdefgh79.21 士6.00 cefghi .Lycopodium casuarinoides Lygodiaceae1.070.002.89 fghijkILygodium flexuosumLygodiacae9.79土 1.63 hjklmn80.652.33 ecdef80.65+2.33 cdefLygodium japonricumLygodiacene21.6726.01 fghij66.67 +6.67 fghijklmno 60.00+5.00 hijklmoMicrosorium buergerianum Polypodiaceae6.67土1.67 ikinnop 86.67土 1.67 bed81.67+6.01 cdefgMicrosorium forruniPolypodiaceae4.17土4.17 mnopqrstu 68.74 19.97 efghijklm 68.74 9.97 fghijklmMicros: puncaum3.95.00+ 2.89 Impopqr88.33土1.67 be85.00土5.00 bed0.0040.00v74.19+3.76 cdefghi74.19土3.76 cdefgbhiNephrolepis cordifolia1.64.77 +0.13 lmnopqrst 73.35 t624 cdefghij73.35 46.24 cdefghijOnychium japoricumSinopteridaceae0.574.19+11.35 a25.81+ 1.35 mnopqrst 25.81土 1.35 opqrsOsmunda japonicaOsmundaceae3.75.03 t 2.76 Imnopqr74.63+5.43 cdefghi74.63+5.43 cefghiOsmunda vachelliOsmundaceac3.4 19.05+0.95 fgijk69.054.97 efhial69.05 土4.97 fghijkimPhymaloptosis hastata83.39+5.88 becde83.39+5.88 bedPinus taiwanensisPinaceaeLeaves and sterms1.756.28+4.21 ecd21.00+3.76 mnopqrst21. 00土3.76 opqrstvar. Daming shanensisPinus yunnaensis.323.83+2.39 efgh69.29 +3.28 efghjl69.29土3.28 ghijklmPodocarpaceae4.67+2.91 Imnopqrs 89.56+1.56 b89.56+1.56 bPseudotaxus chienll80.00+2.89 edefPleridium aquilinumPtenidaceac6.67+中国煤化工Pteris muntifidaPteridaceae4.23.93土80.3 士6.08 cdefgPleris semipinnata11.67土MHCNMHG68.336.01 ghjklmn(Continued on next page)02010,CAAS. A ry red Pltlshedby EieverLud.252HUANG Su-qing et al.Table 1 (Continued from preceding page)YieldMortality")Pupation rate at 72 hEmeragence tateTreatmentFamilyPart(%afier reament(%)(%)无土SE+SEPreris vitataPteridaceaetoots5.00+0.00 Imnopqrst 68.33土 3.33 fghijklmn 68.33 土3.33 ghijklmPreris vittataLeaves8.3.33土1.67 mnopqrstu 68.33+3.33 fghijklmn 66.67+ 1.67 ghijkImSelaginella uncinataSelaginellaceaeWhole plant7.34+3.91 ikmoop69.20+4.60 elghijkl69.20+4.60 (ghjikimTaxus chinensisTaxaceneLeaves and stems4.52+0.29 Imnoprst 60.73 土s.50 hijkmnopq 60.73 $ 5.50 hijklmnoTsugu longibrasteataPinaceaeLeaves and slems 11.0.00+0.00v80.72+2.56 cdef77.94 +4.31 cdefghWoodwardia japonicaBlechnacese9.3 6.67 士4.41 jklmno 81.67+6.01 cder81.67 +6.01 cdefgBlcchnaceaeRoots.6 10.00+0.00 ghijklmn 73.33 1.67 cefgbhij71.67土1.67 fghijk”The crrected morality is the average of three replications士SE, each with 20 adults (n =60).Those wihin a coloumn fllowed by the same ltter aren't sgnificanoly dfferent at 5% level (DMRT). The same as below.tion assays was recorded at 24 and 48 h. Each treat-was the most effective (> 70% mortality). The otherment was replicated 3 times, with 10 flies per replicatefour extracts showed mortalities were higher than 50%(Xu et al.2003).(Table 1). The results also showed that none of these(2) A. albopictus bioassay: Extracts were dissolved withextracts had any impact on the pupation and emergence0.5% acetone. The control was 0.5% acetone alone. Therates of A. albopictus.total number of the tested larvae in each of the treatmentswas 60 with 20 for each of the 3 replicates. The animalsToxicity assay against M. domestica adultswere reared at (25士1)°C. Mortality was assessed 24 hafter treatment. The numbers of pupae and adults wereThe results of the toxicity assay against M. domesticarecorded 3 and 5 d post treatment, respectively.adults are summarized in Table 2. The extracts fromB. orientale (stems), C. funebris (leaves and stems)Statisticsand D. linearis (whole plant) showed high activitieswith mortality rates were higher than 90.00% withinAll the experiments were repeated at least three times24 h after treatment. Mortality rates ranged from 50.00and values were expressed as the means+SE. To evalu-to 89.99% at 24 h after treatment with the extracts ofate the significance of activity differences between theP. taiwanensis var. Daming shanensis (leaves anddata of two chosen groups, the statistical analysis wasstems), P. yunnaensis (leaves and stems), A. muralisperformed by one-way analysis of variance (Duncan's(stems), K. fortunei (barks), K. fortunei (stems),multiple range test) using SAS software. SignificanceO. japonicum (whole plant), A. normale (whole plant), .level was set at P <0.05.and C. spinulosa (leaves). Methanol extracts ofstems), B. orientale (stems), P. yunnaensis (leaves andRESULTSstems), A. muralis (stems), A. muralis (roots), ,K. fortunei (barks), K. fortunei (stems), 0. japonicumToxicity assay against 4th instar larvae of(whole plant), C. funebris (leaves and stems), D. linearisA. albopictus(whole plant), A. normale (whole plant), C. spinulosa(leaves), and O. japonica (whole plant). The mortali-The results of the toxicity assay against 4th instar lar-ties of M. domestica adults were higher than 90% whenvae of A. albopictus are given in Table 1. In the mos-they were treated for 48 h with the extracts ofquito larvicidal assay, the extracts of C. funebris (leavesM. fortune (whole plant), D. griffithii (whole plant),and stems), C. acuminatissima (roots), K. forluneiH. tyermanni (whole plant), C. acuminaissima(leaves and stems), 0. japonicum (whole plant),(ste中国煤化工-oots), C. revoluteand P. taiwanensis var. Daming shanensis (leaves and(root,lant), H. chinensisstems) displayed insecticidal activity at 24 h after (whcMYHCN M H G2 pan),P. hastatetreatment. The extracts of 0. japonicum (whole plant)(whole plant), M. punctatum (whole plant), .02010,CAAS. Al rohss rseved. PlilshedbyEsevherLdAnti-Insetof theMethanol Exracts of Fern and GGymnosperm_253TreatmentomgBL° ain M dmesio adulisParcKMoraliy (%, mean+SE)Abacopers rubra24h afher reamentAdiantum caudatumWhole plant3.33+3.33 nopr48hahetreeatmentAdiantum flbeluaum20.00+5.77 hijki3.33+3.33tAngioplers muralis3.33+3.33 nopqr76.67+6.67 cdeAngioperir muralisStcms10.000.00 Imnop73.33+6.67.de .Angiopters muraliLeaves70.01155540.00士 10.00 ghjkimAspenium 8r/jihinumnRoots16.67*3.33 阔10.00aAspenium rifithianum43.33.3313846.67+8.82 ghijkAspenium normale3.33+3.33 nop90.00+0.00beBlenchnum orientale0.00:0.00 n50.00x5.77 fghiStems3.3x14.53d .33.33+8.82 jkImnoCephalotaxus manni96.673.33 a96.673.33 abChieniopteris hariandi33.33+3.33 fgh0000.000Cibotium barometz60.00土 1.00efghCiborim baromelrSlems16.678.82 jkm46.67*3.33 ghijCpressus Junebris6.67+3.33 mmopq36.67+8.82 ijklmnCyathea podophyllaLeaves and stems12.22+2.22 jkimn46.67+8.82 ghjkCyathea spinulosaWhole piant90.0015.77 ab34.44+2.94 jkimn6.67+6.67 mnop100.00+0.00a73.338.82 cd'16.673.33 nopqCycas acuminatissima13.33.33 jklmo10.00+0.00 aCycas acminaisima50.00土 100fghjCycas acuminisima23.33+8.82 fghj3.0015.28 jImmpCyeas debaoensis16.3613.64私I5.3.33Cycas revolute51.2146.85 fghiPiant10.00+0.00 Imnop36.676.67 jkImnCycar rvolute16.6718.82 ijiklm50.005.77 fghiCelosorus hetercarpusStemns10.00+5.77 kImno63.33.33 rDicranoperis ampla"13.33+8.82 ijklmmo46.67+3.33 ghijDicranopteris dichotomg16.67土12.02 ijklmn26.6716.67 kmnopDicranoperis linearia0.00+0.00nu63.3314.53 deghDicryocline rifuthiiWhole plant .6.67+6.67 mo6.673.33sDrynaria foruneitWholeolant96.673.33343.333 13.33 ghijklmEquisetum difusurm6.67士. 3.33 mnop96.67+3.33 abLaves and stemsFokienia hodeinsi16.67+3.33 i”70.00x0.00cdeHicriopteris chintWhole plan，0.00+0.00m53.33+3.33fgHemauteuchinersi10.00土10.001m30.00土5.77 kmnoHymenophylumpterWhaleplant1.05.7Imp26.67+8.82 klmnopArinoKnepytou brbauom10.0015.77。53.33+6.67ghhole plant! KImnoKeleriafortunei26.67+6.67fe50.00*5.77 fghiLopoarorune76.6728h53.338.82 FghLycopo' calavarumWhole76.673.3310.00.000Lyeadinamn casuarinoidesWhole plat13.33.3m。100.00+0.00 :m. Jlexuosum10.00+53.33 jlmn43.3+6.67 hjkl200577 klmo26.6728.82 kImnoobuergerianum67+3.33 egh，3.33.10p33333 ghij36 678.821mmm puncatum003.33 jklmn266287 jkimn200.0 ji0.01000Meptrolepis cordijolia461333 defgh83.33+10.000 (ghijjaponicum1.00.0 lmop882 bed23.33+8.82 fghiji3333 j75.56+7.29 cd2333+8.82 (ghijensis var. Daming shonensie26.6726.67fghi3.36.6 abe20.00*5.77 hikl5005753.316.67de500*.777 ghi50.00+20.00 cder3.3.333 0ppr90..7.7Pleris munlijida16.73.3343.33土12.02 ghijklmPleris semipinnala中国煤化工3.3.33 nopq30.00*5.77 klmnoMYHCNMHG20.00+5.77 nopqr(Contioued on next page)dbyEererued254HUANG Su-qing etal.Table 2 (Continued from peceding page)Mortality (%，mean+SE)"TreatmentPart24 h after treatment48 h after reatmentPteris vltataRoots30.00士 10.00 fghi73.33+8.82 edePteris witataLeaves23.338.82 fghij66.67+333efSelaginella uncinataWhole plant23.3312.02 ghjkl50.00+5.77 fghiTarus chirensisLeaves and stems16.67+3.33 ijkl43.33 +6.67 hjklTsuga longibrasteata20.0015.77 hijkI40.00士5.77 hijklrdia.aponica23.33+6.67 ghjik46.67 +6.67 ghijikWoodwardia japonica13.33 +8.82 ijkImno40.00土10.00 ghijklm”The crrected mortality is the average of three replicatios土SE, cach with 10 adults (a=30).A. griffthianum (roots), A. caudatum (whole plant)，(whole plant), and O. japonica (whole plant).C. spinulosa (roots), D. fortunei (leaves and stems),A. rubra (whole plant), P. vittata(leaves), P. vittataDISCUSSION(roots), S. uncinata (whole plant), H. barbatum (wholeplant), C. debaoensis (whole plant), and O. vachelliiInsecticidal plants were used to control pests as early(whole plant).as 2000 years ago. Botanical insecticides had been stud-The mortalities of these plant materials were overied systeatically since the 1940s in China and extraor-90% when the applied dose was 10000 mg LI againstdinary progress has been made since the 1980s. Morethe adult of M. domestica (Table 2). However, whenthan 108 commercial botanical insecticides had beenonly a half dose (5000 mg LI) was applied in anotherused in farmlands of China from 1980 to 2009 (Xu andtest (Table 3), none of these extracts displayed activi-Zhang 2009). All commercial botanical insecticides usedties within 24 h after the treatment, while A. muralisin China originated from angiosperms, whereas insec-(roots) showed a mortality of more than 50% at 48 hticidal activities of ferm and gymnosperm were ignoredafter treatment. Mortalities of the tested insects werefor a long time because they are usually not fed by crophigher than 40% at 48 h after exposure to the extractspests (He et al. 2004).of P. taiwanensis var. Daming shanensis (leaves andIn this study, all of the 54 plant species were wildstems), C. acuminatissima (leaves), N. cordifoliafern or gymnosperm collcted from ShiwandashanTable 3 Insecticidal activity of the plant extracts at a dose of 5000 mg LI against M. domestica adultsMortality (%)1_24 h after teatment48 h afer reatment0.00+0.00e3.33.33Abacopteris rubra36.67+8.82 beAngiopteris muralis3.330.21 d56.67+20.28aAniopteris muralis0.0010.00e16.673.33 edeAspenium griffithianum0.00:0.00e16.67-3.33 cdeAspenium normale13.33+3.33 deCupressus funebris16.67+8.82 cdCycas acuminaissima3.33+0.21 d40.00+5.77 beCycas revoluteRools3.33+1.33 gDicranopteris ampla6.67+1.33 g .Dicranopteris dichoroma16.6718.82 edKeteeria fortuneiBarks0.00x0.00e13.33.33 deNephrolepis cordiflia3.33x1.33 d40.00土15.28 abeOnychium japoricum333+1.33d23.33+3.33 cdOsmunda, japonica3.33+1.33 d46.67士12.02 abPinrus taiwas var. Daming shanensis10.0015.7 abPinus yunnaensis0.00+0.00 e0.00+0.00 hPteridium aquilinum10.00+0.00 defPteris vitata中国煤化工6.67+4.67fg30.00+5.28 be .YHCNMHG30.00土1.55 be ."The coreted mortality is the average of three replications t SE, each with 10 aduts (n=30).02010.CAS. AI nohe ree lbloyoweredAnti-Insect Activity of the Methanol Extracts of Fern and Gymnosperm255Mountain, Guangxi Zhuang Autonomous Region into adult houseflies (aking 40.74%, 22/54), and extractsSouthwest China. Biological activity of the extracts offrom 5 plants are toxic to the 4th larvae of mosquitoesmost of the tested plants against bacteria, fungi and(taking 9.26%, 5/54). This indicates that ferns andviruses had been reported. The bioactivities of the 7gymnosperms possess obvious insecticidal actities toplants included in this study had been reported by othernon-addicted feeding insects and could be a preciousauthors (Zhou and Li 1999; Xu 2005; Xu et al. 2005).library of lead compounds with insecticidal activity.Water extracts of Selaginella inrolrens, S. uncinata,From ferns and gymnosperms, it is possible to findO. japonica, L. japonicum, Stenoloma chusamum,compounds which originated from the ancient time andP. muntifda, and Lepidogranmis drymoglssoides pssessed acivities to inets of Late Mesozoic, andshowed remarkable antibacterial activity against Shi-these compounds may have marvellous effects in pro-gella sp., Escherichia coli, Proleus rulgaris, Staphylo-tecting crops.coccus aureus, and Bacterium pyocyaneum (Zhou andIn this paper, the bioassay showed that many metha-Li 1999; Kubo et al. 1992). It was also documentednol extracts of the fem or gymnosperm possessed ob-that polysaccharide extracts of D. dichotoma, S. chusana,vious bioactivity against A. albopictus and M. domestica.P. aquilinum, P. vittata, P. multifida, O. japonicum,It provides a potential to find new insecticidal com-Adiantum, flabellulatu, and Cyclosorus parasiticus ex-pound from fern or gymnosperm. They are rich inhibited obvious inhibitory effects against bacteria andSouth China and the roots are shallower, which makesfungi (Xu et al. 2005). But study of insecticidal com-collection easy. Therefore they are ideal materials topounds from the pteridophyte and gymnosperm hasproduce botanical insecticides.not been undertaken.Angiosperms are currently dominant on the earth,Acknowledgementsand hurted by major insects which have important im-This research was funded by the National Nature Sci-pact on the economical profits of the human being.ence Foundation of China (30571235) and the IndustryThe study of relationship between insects and plantsSchemes of the Ministry of Agriculture, Chinahas been generally focused on the relationship between(200903052). The authors greatly thank Professor Zenginsects and angiosperms, especially between theDongqiang of Guangxi University, China, for his help .lepidopteron which originated from the Late Cretaceousin plant material collection and Professor Zhong Yecongand angiosperms. The relationship between ferns orfor his help in identification of the plant specimen.gymnosperms and the agricultural insects is not inti-mate and study of the relationship between them hasReferencesnot been emphasized, which is especially manifestedEditorial Committee of Flora of China, Chinese Academy ofby the study of botanical insecticides. So far, the ac-Sciences. 1978. Flora of China. vol.7. Science Press, Beijing.tive ingredients in botanical insecticides all come from(in Chinese)angiosperms. The co evolution history of fems or gym-He TP, Wen Y G, Wen X F, LiJ G.2004. The plant resources innosperms and insects is far longer than that of an-Shiwandadashan Mountain natural reserve of Guangxi.giosperms and insects (Editorial Commite of Flora ofChinese Wild Plant Resources, 1, 23-26. (in Chinese)China, Chinese Academy of Sciences 1978). In theKubo I, Murol H, Himejima M.1992. Antibacterial activity oftotarol and its potentiation. 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