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1SCiEntifiC RePoRTS | (2018)8:10565

Microemulsion formulation of a

new biopesticide to control the diamondback moth (Lepidoptera:

Plutellidae)

Hainan

Shao, Na Xi & Yalin Zhang

This study was designed to develop a microemulsion formulation of norcantharidin for the control of the diamondback moth (DBM),

Plutella xylostella

(Linnaeus), a notorious pest of brassica crops worldwide. The oil phase was screened and selected based on norcantharidin solubility while the for norcantharidin microemulsion to P . xylostella P. xylostella and possibly contribute to ameliorating any pesticide resistance in P . xylostella. drawnwideattentionduringrecentyears1 vehiclesbecauseoftheiruniqueadvantages 2-5 nm)andbeingcompletely water-dilutable 6,7 .?issmalldropletsizeandlowered traditionalcommercialpesticides3,7,9 .Meanwhile,ithasbeenreportedthat

λ-cyhalothrinandtriadimefonmicro-

10 ,11 12

14.Applicationofsyntheticinsecti-

15 .Insectileresistanceandcross-resistance

Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, College of Plant

)Received: 15 March 2018 A ccepted: 18 June 2018 P ublished: xx xx xxxx www.nature.com/scientificreports/

2SCiEntifiC RePoRTS | (2018)8:10565

selectingforanyspeci?ctypeofresistance. invivo 16 .Proteinserine/threoninephosphatasesplay larityineukaryoticcells 17 .Chen threoninephosphatasegenesof P .xylostellainvitro 18 controlandagainstpathogenicfungi 19 ,20 radationrateandnewmodeofaction 21
e?ectiveandsafepesticidetocontrol analyzedagainst3rdinstarsof P

Results and Discussion

thetoxicityofthesystem 22
g/Lwhen contentofnorcantharidinwasmaintainedat50 23
.Itwasreportedthatmixedsurfactants asinglephase 24
25
,26 inTable 1,Tx13andTw80werethemostappropriatesurfactantsduetoformingasinglecontinuousphasewitha

1:1providinggoodthermodynamicstability(Table 2).Non-ionicsurfactantsareo?enusedforthescreening

22
systembyinsertionintotheinterfaciallayer 27
23
,28 .?ee?ectsofdi?erent surfactants-to-cosurfactantratios(Km microemulsion 29
.AsshowninFig. 1,themicroemulsionregionschangewiththeadditionofdi?erentratiosofcosu- rfactant.Forthemono-phaseregion,thesmallest(32.75%)wasformedwhenKmwas4:1(Fig. 1A),whereasthe largest(39.47%)wasdetectedwhenKmreached4:4(Fig. 1D),suggestingthat4:4wasthemostappropriateKmto andethylbutyrate(1:1:2),15%(w/w)Tw80andTx13(1:1),15%(w/w)ethanol,and58%(w/w)water(Table 3).

Conductivityisakeytechniqueto

30

ture.Obviously,fourregionsexistasindicatedinFig. 2.?eelectricalconductivityσoftheselectedoilymixture

fraction( w )reachedacriticalpoint( p www.nature.com/scientificreports/

3SCiEntifiC RePoRTS | (2018)8:10565

30
,31 .Weascribethelinearincreasein 27
reachesthemaximumvalue(1.25 Sm 1 w 23
,32 33
.?estructureofthemicroemulsionfromw/otoa discontinuousphaseandtheno/wusingtitrationisindicatedbythederivativeoftheconductivity(Fig. 2).

Stability of microemulsions.

ductedusingfreeze-thawcyclesbetween lessthan48 aturegreaterthan72 34
catedanincreaseinthedropletsizealongwiththeaugmenteddilutionratio(Fig. 3).?eincreaseofdropletradii radiiofdilutedsamples 34
7,35

SurfactantsAppearanceAppearancea?er24h

Tw80TransparentsolutionTransparent

Tx13TransparentsolutionTransparent

Table1.

Assayofscreeningoptimalsurfactants.

3:1Turbid - - - -

3:2Turbid - - - -

2:1TransparentTransparentSomeprecipitation - -

2:3TransparentTurbid - - -

Table2.

www.nature.com/scientificreports/

4SCiEntifiC RePoRTS | (2018)8:10565

Insecticidalactivity.

ticidalandanti-fungalactivities 19 ,20 e?ciency.AccordingtoTable 4,all6commercialinsecticidesareshowntobehighlyactiveagainstP.xylostella 50
(0.07 mg/L;0.06-0.09,95%CL).Even 50
(6.501and12.477 mg/L)toearly-stagelarvaeofP. 21
procedures 36
,37 aswellasscarceorigin 38
h LC 50

P.xylostellawasfoundtobe

12.477(11.58-13.41)mg/L.

P .xylostellathandidtheirpure 50
,respectively)(Table 4).Previousstudies correspondingactiveingredients 39
,40 41
.Lei etal.(2012,2014)and

Oil/SAA/H

2

Table3.

Figure1

2

Osystematdi?erentratiosofemulsi?ers-to-

cosurfactant,( A )Km =4:1,(B)Km=4:2,(C)Km=4:3,(D)Km=4:4. www.nature.com/scientificreports/

5SCiEntifiC RePoRTS | (2018)8:10565

formulations 3,7,9 3 muchbetteractivitythanacorrespondinglypureactiveingredientalone(Table 4). P. xylostella(Table 5),especiallyspinosadandemamectinbenzoatewhichshowedthehighesttoxicities,withalar- vaedeathrateupto100%within24 instarswithin24 toxicitytoearly-stagelarvaeof P .xylostella 42
43
44
45
e.g.,spinosad,indoxacradandemamectinben zoate 42
46
47
areeco-friendly 48
49
52

Figure2

mixedoil/watersystem(standardwater,342

Figure3

dilutiontimeswithstandardwater(342 mg/L). www.nature.com/scientificreports/

6SCiEntifiC RePoRTS | (2018)8:10565

Materials and Methods

conditions(25

Norcantharidin(

TechnologiesCo.,Ltd),60

Solvent screening.

solventswasalsomeasured.0.5 waschoseforfurtherexperiments. Surfactant screening. Inordertoscreenandselectthemoste?ectivesurfactant(s),mixedsolutionsof 20 formulation.

InsecticidesConcentration

12 h24h48h

Control0.00.00.0

Cantharidin1534.5±1.48

b 82.3

±0.78

b 100
a

Norcantharidin3021.3±0.83

c 56.8

±0.45

c 95.6

±3.24

a

Azadirachtin10.535.9±1.02

b 56.4

±1.26

c 100
a

Emamectinbenzoate590.0±0.51

a 95.0

±3.14

a 100
a

Indoxacarb4512.5±0.12

d 52.5

±5.47

c 97.5

±3.15

a

Spinosad3092.5±0.05

a 100
a 100
a

Table5.

P <0.05).

InsecticidesFormulationActive

ingredients(%)n a

Slope(

±SE)Relationcoe?cientLC

50
mg/L(95%CL b 2

Table4.

P .xylostellainaleaf-dipbioassay(48h). a

Numberoflarvaetested,

b

Con?denceinterval;EC:emulsi?able

www.nature.com/scientificreports/

7SCiEntifiC RePoRTS | (2018)8:10565

China)at25

inversionofthemicroemulsionfromO/WtoW/O. water(contentofCa 2 ,Mg 2 were342

°C.

Centrifugation.

outtoassessthephysicalstabilityofME. ?ermodynamicstability. and54 measured.Meanwhile,sampleswerestoredat increaseat2.0 53
m×0.53mm×1.5 ml/min),hydrogenat2.0kg/cm 2 andairat2.55 kg/ cm 2 .?einitialoventemperaturewas150 followingformula: RM100 RM

Concentration(%)

21
12

Where:R

1 R 2 :ratioofthe M 1 :theweightofstandardnorcantharidin (g);M 2

Bioassays.

takenasacontrol.Leafdiscs(Ø1.0 about10quotesdbs_dbs41.pdfusesText_41
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