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Ernesto Gabriel Fuentes, Estrella Hernández-Suárez, Oihane Simón, Trevor Williams, Primitivo Caballero
Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV): Natural occurrence and efficacy as a biological insecticide on young banana plants in greenhouse and open-field conditions on the Canary Islands
www.plosone.org
www.plosone.org
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(As)Tj
1.2643 0 Td
(the)Tj
1.4683 0 Td
(density)Tj
-34.7128 -1.2983 Td
(of)Tj
1.0147 0 Td
(infestation)Tj
4.4618 0 Td
(varied,)Tj
2.9196 0 Td
(the)Tj
1.474 0 Td
(number)Tj
3.4186 0 Td
(of)Tj
1.0148 0 Td
(larvae)Tj
2.5795 0 Td
(collected)Tj
3.7417 0 Td
(at)Tj
0.9524 0 Td
(each)Tj
2.0296 0 Td
(site)Tj
1.5704 0 Td
(varied)Tj
2.6986 0 Td
(accordingly)Tj
4.8869 0 Td
(\()Tj
0.83 0.64 0.02 0 k
(Table)Tj
2.7836 0 Td
(1)Tj
0 g
(\).)Tj
-34.35 -1.3039 Td
(Larvae)Tj
2.8799 0 Td
(were)Tj
2.1203 0 Td
(collected)Tj
3.7418 0 Td
(using)Tj
2.3867 0 Td
(a)Tj
0.6576 0 Td
(soft)Tj
1.6725 0 Td
(paintbrush,)Tj
4.8018 0 Td
(and)Tj
1.7348 0 Td
(individualized)Tj
5.9187 0 Td
(in)Tj
1.0318 0 Td
(25)Tj
1.1736 0 Td
(ml)Tj
1.2869 0 Td
(plastic)Tj
2.7666 0 Td
(pots)Tj
1.9049 0 Td
(con-)Tj
-35.2741 -1.2983 Td
(taining)Tj
3.05 0 Td
(diet.)Tj
1.9616 0 Td
(Larvae)Tj
2.8743 0 Td
(were)Tj
2.1203 0 Td
(maintained)Tj
4.7905 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.6519 0 Td
(laboratory)Tj
4.3484 0 Td
(rearing)Tj
3.101 0 Td
(chamber)Tj
3.7304 0 Td
(at)Tj
0.9525 0 Td
(25)Tj
/F13 1 Tf
1.1678 0 Td
[()]TJ
/F7 1 Tf
0.7937 0 Td
(2C,)Tj
1.9729 0 Td
(70)Tj
/F13 1 Tf
1.1736 0 Td
[()]TJ
/F7 1 Tf
0.788 0 Td
(15%)Tj
-34.5144 -1.2983 Td
(RH)Tj
1.61 0 Td
(and)Tj
1.7291 0 Td
(16:8)Tj
1.8709 0 Td
(hours)Tj
2.5171 0 Td
(\(L:D\))Tj
2.3981 0 Td
(photoperiod,)Tj
5.4142 0 Td
(until)Tj
2.1089 0 Td
(death)Tj
2.4322 0 Td
(or)Tj
1.0998 0 Td
(pupation.)Tj
4.0592 0 Td
(Larvae)Tj
2.88 0 Td
(were)Tj
2.1203 0 Td
(inspected)Tj
4.0081 0 Td
(daily)Tj
-34.2479 -1.3039 Td
(and)Tj
1.7291 0 Td
(those)Tj
2.3357 0 Td
(died)Tj
1.9672 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.474 0 Td
(typical)Tj
2.8573 0 Td
(signs)Tj
2.2224 0 Td
(of)Tj
1.0148 0 Td
(polyhedrosis)Tj
5.2951 0 Td
(disease)Tj
3.016 0 Td
(were)Tj
2.1203 0 Td
(individually)Tj
4.9889 0 Td
(frozen)Tj
2.7723 0 Td
(at)Tj
0.9524 0 Td
(-20C)Tj
-34.7411 -1.2982 Td
(for)Tj
1.3832 0 Td
(subsequent)Tj
4.7055 0 Td
(analysis.)Tj
3.5547 0 Td
(All)Tj
1.3946 0 Td
(larvae)Tj
2.5852 0 Td
(that)Tj
1.7801 0 Td
(died)Tj
1.9616 0 Td
(were)Tj
2.126 0 Td
(observed)Tj
3.7927 0 Td
(using)Tj
2.3924 0 Td
(an)Tj
1.2019 0 Td
(optical)Tj
2.914 0 Td
(microscope)Tj
-29.7919 -1.2983 Td
(\(x1000\))Tj
3.2654 0 Td
(to)Tj
1.0262 0 Td
(determine)Tj
4.3256 0 Td
(the)Tj
1.4684 0 Td
(presence)Tj
3.7133 0 Td
(of)Tj
1.0148 0 Td
(OBs.)Tj
-13.6175 -1.3039 Td
(To)Tj
1.3379 0 Td
(determine)Tj
4.3256 0 Td
(the)Tj
1.4684 0 Td
(identity)Tj
3.3052 0 Td
(of)Tj
1.0148 0 Td
(isolates)Tj
3.118 0 Td
(from)Tj
2.2054 0 Td
(virus-killed)Tj
4.7622 0 Td
(field-collected)Tj
5.845 0 Td
(insects,)Tj
3.1237 0 Td
(restriction)Tj
-31.7024 -1.2983 Td
(endonuclease)Tj
5.6125 0 Td
(analysis)Tj
3.3279 0 Td
(\(REN\))Tj
2.8289 0 Td
(was)Tj
1.6951 0 Td
(performed.)Tj
4.6942 0 Td
(For)Tj
1.6271 0 Td
(this,)Tj
1.8992 0 Td
(viral)Tj
1.9955 0 Td
(occlusion)Tj
4.0309 0 Td
(bodies)Tj
2.8119 0 Td
(\(OBs\))Tj
2.5909 0 Td
(were)Tj
2.1203 0 Td
(puri-)Tj
-35.2344 -1.2982 Td
(fied)Tj
1.7234 0 Td
(from)Tj
2.2053 0 Td
(cadavers)Tj
3.651 0 Td
(by)Tj
1.1849 0 Td
(homogenization)Tj
6.7521 0 Td
(in)Tj
1.0318 0 Td
(0.1%)Tj
2.1543 0 Td
(\(wt/vol\))Tj
3.4243 0 Td
(sodium)Tj
3.2258 0 Td
(dodecyl)Tj
3.3335 0 Td
(sulfate)Tj
2.8006 0 Td
(\(SDS\))Tj
2.5852 0 Td
(and)Tj
-34.0722 -1.2983 Td
(were)Tj
2.1203 0 Td
(filtered)Tj
3.067 0 Td
(through)Tj
3.4469 0 Td
(muslin)Tj
2.9877 0 Td
(to)Tj
1.0262 0 Td
(remove)Tj
3.2201 0 Td
(debris.)Tj
2.9027 0 Td
(The)Tj
1.7801 0 Td
(resulting)Tj
3.7304 0 Td
(suspension)Tj
4.6431 0 Td
(was)Tj
1.7008 0 Td
(centrifuged)Tj
4.7848 0 Td
(at)Tj
-35.4101 -1.3039 Td
(3,800)Tj
2.347 0 Td
(x)Tj
/F12 1 Tf
0.686 0 Td
[()]TJ
/F7 1 Tf
0.6293 0 Td
(for)Tj
1.389 0 Td
(5)Tj
0.6973 0 Td
(minutes,)Tj
3.6907 0 Td
(OBs)Tj
1.9048 0 Td
(were)Tj
2.126 0 Td
(resuspended)Tj
5.2327 0 Td
(in)Tj
1.0318 0 Td
(500)Tj
/F13 1 Tf
1.6498 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9694 0 Td
(of)Tj
1.0148 0 Td
(0.1%)Tj
2.1543 0 Td
(SDS)Tj
1.9106 0 Td
(and)Tj
1.7291 0 Td
(centrifuged)Tj
4.7849 0 Td
(again)Tj
2.3697 0 Td
(at)Tj
-36.3172 -1.2983 Td
(3,800)Tj
2.347 0 Td
(x)Tj
/F12 1 Tf
0.686 0 Td
[()]TJ
/F7 1 Tf
0.6293 0 Td
(for)Tj
1.389 0 Td
(5)Tj
0.6973 0 Td
(minutes.)Tj
3.6907 0 Td
(OBs)Tj
1.9048 0 Td
(were)Tj
2.126 0 Td
(finally)Tj
2.7042 0 Td
(resuspended)Tj
5.2328 0 Td
(in)Tj
1.0318 0 Td
(sterile)Tj
2.6135 0 Td
(distilled)Tj
3.3845 0 Td
(water.)Tj
2.6589 0 Td
(For)Tj
1.6214 0 Td
(viral)Tj
1.9956 0 Td
(DNA)Tj
-34.7128 -1.2983 Td
(extraction,)Tj
4.4787 0 Td
(100)Tj
/F13 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9751 0 Td
(of)Tj
1.0148 0 Td
(purified)Tj
3.4072 0 Td
(OB)Tj
1.5478 0 Td
(suspension)Tj
4.6431 0 Td
(was)Tj
1.6951 0 Td
(mixed)Tj
2.7269 0 Td
(with)Tj
1.9956 0 Td
(100)Tj
/F13 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9695 0 Td
(of)Tj
1.0148 0 Td
(0.5M)Tj
2.2903 0 Td
(Na)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 512.2771 445.7196 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 515.8488 447.4204 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 529.9653 445.7196 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 533.537 447.4204 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4422 0 Td
(50)Tj
/F13 1 Tf
1.1735 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9694 0 Td
(of)Tj
-35.9373 -1.3039 Td
(10%)Tj
1.9276 0 Td
(SDS)Tj
1.9048 0 Td
(and)Tj
1.7348 0 Td
(distilled)Tj
3.3846 0 Td
(water)Tj
2.4321 0 Td
(to)Tj
1.0205 0 Td
(a)Tj
0.6576 0 Td
(final)Tj
2.0012 0 Td
(volume)Tj
3.1975 0 Td
(of)Tj
1.0148 0 Td
(500)Tj
/F13 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(l,)Tj
1.1963 0 Td
(incubated)Tj
4.1782 0 Td
(at)Tj
0.9524 0 Td
(60C)Tj
2.228 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1736 0 Td
(minutes)Tj
3.4638 0 Td
(and)Tj
-35.5008 -1.2982 Td
(centrifuged)Tj
4.7849 0 Td
(at)Tj
0.9524 0 Td
(3,800)Tj
2.3471 0 Td
(x)Tj
/F12 1 Tf
0.686 0 Td
[()]TJ
/F7 1 Tf
0.6292 0 Td
(for)Tj
1.389 0 Td
(5)Tj
0.6973 0 Td
(minutes.)Tj
3.6907 0 Td
(The)Tj
1.7802 0 Td
(resulting)Tj
3.7303 0 Td
(supernatant)Tj
4.9833 0 Td
(was)Tj
1.6951 0 Td
(treated)Tj
2.9991 0 Td
(with)Tj
1.9955 0 Td
(25)Tj
/F13 1 Tf
1.1735 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9751 0 Td
(of)Tj
1.0148 0 Td
(pro-)Tj
-35.5235 -1.2983 Td
(teinase)Tj
2.965 0 Td
(K)Tj
0.8901 0 Td
(\(20)Tj
1.5024 0 Td
(mg/ml\))Tj
3.2314 0 Td
(at)Tj
0.9468 0 Td
(50C)Tj
2.2167 0 Td
(for)Tj
1.3776 0 Td
(30)Tj
1.1679 0 Td
(minutes)Tj
3.4582 0 Td
(and)Tj
1.7235 0 Td
(then)Tj
2.0069 0 Td
(treated)Tj
2.9934 0 Td
(twice)Tj
2.3074 0 Td
(with)Tj
1.9899 0 Td
(500)Tj
/F13 1 Tf
1.6384 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9637 0 Td
(of)Tj
1.0092 0 Td
(TE-saturated)Tj
-32.3885 -1.3039 Td
(phenol)Tj
2.9877 0 Td
(and)Tj
1.7291 0 Td
(once)Tj
2.126 0 Td
(with)Tj
1.9956 0 Td
(500)Tj
/F13 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9695 0 Td
(chloroform.)Tj
5.0173 0 Td
(The)Tj
1.7801 0 Td
(aqueous)Tj
3.5093 0 Td
(phase)Tj
2.4831 0 Td
(was)Tj
1.7008 0 Td
(recovered)Tj
4.1442 0 Td
(and)Tj
1.7292 0 Td
(DNA)Tj
2.3753 0 Td
(was)Tj
1.7007 0 Td
(pre-)Tj
-35.8976 -1.2983 Td
(cipitated)Tj
3.6737 0 Td
(by)Tj
1.1849 0 Td
(adding)Tj
2.9877 0 Td
(1/10)Tj
1.9785 0 Td
(volume)Tj
3.1975 0 Td
(3M)Tj
1.5874 0 Td
(sodium)Tj
3.2315 0 Td
(acetate)Tj
2.9536 0 Td
(at)Tj
0.9525 0 Td
(pH)Tj
1.508 0 Td
(5.2)Tj
1.3946 0 Td
(and)Tj
1.7292 0 Td
(2.5)Tj
1.4003 0 Td
(volumes)Tj
3.5546 0 Td
(of)Tj
1.0205 0 Td
(100%)Tj
2.4036 0 Td
(cold)Tj
-34.7581 -1.2982 Td
(ethanol.)Tj
3.4299 0 Td
(The)Tj
1.7802 0 Td
(DNA)Tj
2.3811 0 Td
(pellet)Tj
2.3697 0 Td
(was)Tj
1.7008 0 Td
(washed)Tj
3.1804 0 Td
(with)Tj
1.9956 0 Td
(70%)Tj
1.9276 0 Td
(cold)Tj
1.9275 0 Td
(ethanol)Tj
3.2031 0 Td
(and)Tj
1.7292 0 Td
(resuspended)Tj
5.2327 0 Td
(in)Tj
1.0375 0 Td
(50100)Tj
/F13 1 Tf
3.1123 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
-35.0076 -1.2983 Td
(0.1x)Tj
1.8595 0 Td
(TE)Tj
1.4003 0 Td
(\(Tris)Tj
2.1714 0 Td
(HCl-EDTA\))Tj
5.1873 0 Td
(buffer.)Tj
2.8517 0 Td
(For)Tj
1.627 0 Td
(REN)Tj
2.1487 0 Td
(analysis,)Tj
3.5489 0 Td
(2)Tj
/F13 1 Tf
0.6974 0 Td
[()]TJ
/F7 1 Tf
(g)Tj
1.1905 0 Td
(of)Tj
1.0148 0 Td
(viral)Tj
1.9956 0 Td
(DNA)Tj
2.3754 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1781 0 Td
(with)Tj
-34.3669 -1.3039 Td
(5)Tj
0.6917 0 Td
(units)Tj
2.228 0 Td
(of)Tj
/F12 1 Tf
1.0148 0 Td
[()-2()]TJ
/F7 1 Tf
(II)Tj
2.1146 0 Td
(\(Takara\))Tj
3.685 0 Td
(at)Tj
0.9468 0 Td
(37C)Tj
2.228 0 Td
(for)Tj
1.389 0 Td
(4)Tj
0.6916 0 Td
(hours)Tj
2.5172 0 Td
(following)Tj
3.9798 0 Td
(the)Tj
1.4683 0 Td
(manufacturer's)Tj
6.2646 0 Td
(instructions.)Tj
5.2269 0 Td
(The)Tj
-34.4463 -1.2983 Td
(reaction)Tj
3.4923 0 Td
(was)Tj
1.7008 0 Td
(stopped)Tj
3.3675 0 Td
(by)Tj
1.1849 0 Td
(adding)Tj
2.9877 0 Td
(4)Tj
/F13 1 Tf
0.6973 0 Td
[()]TJ
/F7 1 Tf
(l)Tj
0.9751 0 Td
(of)Tj
1.0148 0 Td
(loading)Tj
3.2145 0 Td
(buffer)Tj
2.6248 0 Td
(\(0.25%)Tj
2.9707 0 Td
(w/v)Tj
1.6895 0 Td
(bromophenol)Tj
5.6976 0 Td
(blue,)Tj
2.1542 0 Td
(40%)Tj
1.9275 0 Td
(w/v)Tj
-35.6992 -1.2983 Td
(sucrose\).)Tj
3.7587 0 Td
(Samples)Tj
3.498 0 Td
(were)Tj
2.1203 0 Td
(loaded)Tj
2.8856 0 Td
(on)Tj
1.2756 0 Td
(1%)Tj
1.457 0 Td
(agarose)Tj
3.2145 0 Td
(gel)Tj
1.3549 0 Td
(and)Tj
1.7291 0 Td
(subjected)Tj
3.9685 0 Td
(to)Tj
1.0262 0 Td
(electrophoresis)Tj
6.2475 0 Td
(in)Tj
1.0374 0 Td
(TAE)Tj
-33.5733 -1.3039 Td
(buffer)Tj
2.6249 0 Td
(\(40)Tj
1.508 0 Td
(mM)Tj
1.9332 0 Td
(Tris-acetate,)Tj
5.142 0 Td
(pH)Tj
1.5081 0 Td
(8.0;)Tj
1.6214 0 Td
(1)Tj
0.6916 0 Td
(mM)Tj
1.9332 0 Td
(EDTA\).)Tj
3.3902 0 Td
(The)Tj
1.7802 0 Td
(gel)Tj
1.3549 0 Td
(was)Tj
1.7008 0 Td
(stained)Tj
3.0784 0 Td
(using)Tj
2.3925 0 Td
(ethidium)Tj
3.8776 0 Td
(bro-)Tj
-34.537 -1.2982 Td
(mide,)Tj
2.4832 0 Td
(observed)Tj
3.7927 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
0.6576 0 Td
(UV)Tj
1.6498 0 Td
(transilluminator)Tj
6.7634 0 Td
(and)Tj
1.7348 0 Td
(photographed)Tj
5.8393 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
1.4684 0 Td
(GeneSnap)Tj
4.3086 0 Td
(Chemi-Doc)Tj
-32.3658 -1.2983 Td
(package)Tj
3.4129 0 Td
(\(BioRad,)Tj
3.7247 0 Td
(CA\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 242.9858 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Production)Tj
4.9417 0 Td
(of)Tj
1.0441 0 Td
(ChchNPV-TF1)Tj
6.6709 0 Td
(OBs)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 225.9779 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(For)Tj
1.627 0 Td
(field)Tj
1.9729 0 Td
(trials,)Tj
2.4208 0 Td
(ChchNPV-TF1)Tj
6.3156 0 Td
(isolate)Tj
2.7609 0 Td
(OBs)Tj
1.9048 0 Td
(were)Tj
2.1203 0 Td
(produced)Tj
4.0479 0 Td
(by)Tj
1.1792 0 Td
(inoculating)Tj
4.7565 0 Td
(fifth)Tj
1.8879 0 Td
(and)Tj
1.7291 0 Td
(sixth)Tj
2.143 0 Td
(instar)Tj
-34.8659 -1.2982 Td
(laboratory-reared)Tj
/F12 1 Tf
7.2509 0 Td
[()]TJ
/F7 1 Tf
(.)Tj
/F12 1 Tf
1.0658 0 Td
[(\n)13()6()11()4(\n)1()14(\r)11()8()]TJ
/F7 1 Tf
3.481 0 Td
(larvae)Tj
2.5852 0 Td
(with)Tj
1.9955 0 Td
(5.00x10)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 394.4692 217.4173 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(7)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 400.2519 212.9952 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(OBs/ml)Tj
3.2995 0 Td
(or)Tj
1.0998 0 Td
(9.02x10)Tj
ET
Q
q
1 j
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1 j
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dup 115 /s put
dup 116 /t put
dup 117 /u put
dup 118 /v put
dup 119 /w put
dup 120 /x put
dup 121 /y put
dup 122 /z put
dup 123 /braceleft put
dup 124 /bar put
dup 125 /braceright put
dup 126 /asciitilde put
readonly def
currentdict end
currentfile eexec
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