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ArticleTitle Effect of trichomes on the predation of Tetranychus urticae (Acari: Tetranychidae) by Phytoseiulusmacropilis (Acari: Phytoseiidae) on tomato, and the interference of webbing

Article Sub-Title

Article CopyRight Springer Science+Business Media B.V.(This will be the copyright line in the final PDF)

Journal Name Experimental and Applied Acarology

Corresponding Author Family Name SatoParticle

Given Name Monica M.Suffix

Division Departamento de Entomologia e Acarologia

Organization ESALQ-USP

Address Piracicaba, SP, 13418-900, Brazil

Email mmsato@esalq.usp.br

Author Family Name MoraesParticle deGiven Name Gilberto J.Suffix

Division Departamento de Entomologia e Acarologia

Organization ESALQ-USP

Address Piracicaba, SP, 13418-900, Brazil

Email gjmoraes@carpa.ciagri.usp.br

Author Family Name HaddadParticle

Given Name Marineia L.Suffix

Division Departamento de Entomologia e Acarologia

Organization ESALQ-USP

Address Piracicaba, SP, 13418-900, Brazil

Email mlhaddad@esalq.usp.br

Author Family Name WekesaParticle

Given Name Vitalis W.Suffix

Division Departamento de Entomologia e Acarologia

Organization ESALQ-USP

Address Piracicaba, SP, 13418-900, Brazil

Email ewekesah@yahoo.com

Schedule

Received 30 July 2009

Revised

Accepted 17 January 2011

Abstract The two-spotted spider mite, Tetranychus urticae Koch, is an important pest of tomato in different parts ofthe world. Biological control of this pest on this crop has not been very successful on this crop.Phytoseiulus macropilis (Banks) has been used commercially for the control of T. urticae on different crops,but no information has been published on its potential to control T. urticae on tomato. The objective of thiswork was to compare the performance of a Brazilian population of P. macropilis on tomato with itsperformance on other plant species, relating the observed variation to the respective types and densities oftrichomes. It has been hypothesized that the presence of the webbing produced by T. urticae could help thepredator to avoid contact with trichomes and consequently to improve its performance on tomato plants. Thishypothesis was also evaluated. Phytoseiulus longipes Evans was included in the work as a control, given thatit has been reported to be a promising predator of Tetranychus Boisduval species on tomato. The study wasconducted under laboratory conditions. It was found that the performance of P. macropilis was similar to thatof P. longipes and that trichomes hampered the locomotion as well as prey consumption and oviposition rateof both predators; that the presence of webbing resulted in higher levels of prey consumption and of predatoroviposition; and that the presence of webbing eliminated the negative effect of trichomes of eggplants andpartially eliminated the negative effect of trichomes of the ‘cerasiforme’ tomato variety. The observed densityof trichomes in the ‘Carmem’ tomato variety, one of the most common tomato varieties grown in Brazil,seems not to have interfered significantly with the prey consumption and the oviposition rate of P.macropilis. The results suggest that the latter is a promising predator of T. urticae on tomato. Complementarystudies are warranted, to further evaluate the potential of P. macropilis for use as a biological control agentof this pest.

Keywords (separated by '-') Biological control - Predatory mites - Trichomes - SolanaceaeFootnote Information

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12

3 Effect of trichomes on the predation of Tetranychus

4 urticae (Acari: Tetranychidae) by Phytoseiulus macropilis

5 (Acari: Phytoseiidae) on tomato, and the interference

6 of webbing

7 Monica M. Sato • Gilberto J. de Moraes • Marineia L. Haddad •

8 Vitalis W. Wekesa

9 Received: 30 July 2009 / Accepted: 17 January 201110 � Springer Science+Business Media B.V. 2011

11 Abstract The two-spotted spider mite, Tetranychus urticae Koch, is an important pest of

12 tomato in different parts of the world. Biological control of this pest on this crop has not

13 been very successful on this crop. Phytoseiulus macropilis (Banks) has been used com-

14 mercially for the control of T. urticae on different crops, but no information has been

15 published on its potential to control T. urticae on tomato. The objective of this work was to

16 compare the performance of a Brazilian population of P. macropilis on tomato with its

17 performance on other plant species, relating the observed variation to the respective types

18 and densities of trichomes. It has been hypothesized that the presence of the webbing

19 produced by T. urticae could help the predator to avoid contact with trichomes and con-

20 sequently to improve its performance on tomato plants. This hypothesis was also evalu-

21 ated. Phytoseiulus longipes Evans was included in the work as a control, given that it has

22 been reported to be a promising predator of Tetranychus Boisduval species on tomato. The

23 study was conducted under laboratory conditions. It was found that the performance of

24 P. macropilis was similar to that of P. longipes and that trichomes hampered the loco-

25 motion as well as prey consumption and oviposition rate of both predators; that the

26 presence of webbing resulted in higher levels of prey consumption and of predator ovi-

27 position; and that the presence of webbing eliminated the negative effect of trichomes of

28 eggplants and partially eliminated the negative effect of trichomes of the ‘cerasiforme’

29 tomato variety. The observed density of trichomes in the ‘Carmem’ tomato variety, one of

30 the most common tomato varieties grown in Brazil, seems not to have interfered signifi-

31 cantly with the prey consumption and the oviposition rate of P. macropilis. The results

32 suggest that the latter is a promising predator of T. urticae on tomato. Complementary

A1 M. M. Sato (&) � G. J. de Moraes � M. L. Haddad � V. W. Wekesa

A2 Departamento de Entomologia e Acarologia, ESALQ-USP, Piracicaba, SP 13418-900, Brazil

A3 e-mail: mmsato@esalq.usp.br

A4 G. J. de Moraes

A5 e-mail: gjmoraes@carpa.ciagri.usp.br

A6 M. L. Haddad

A7 e-mail: mlhaddad@esalq.usp.br

A8 V. W. Wekesa

A9 e-mail: ewekesah@yahoo.com

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33 studies are warranted, to further evaluate the potential of P. macropilis for use as a

34 biological control agent of this pest.

35 Keywords Biological control � Predatory mites � Trichomes � Solanaceae

36 Introduction

37 The two-spotted spider mite (Tetranychus urticae Koch) is one of the main tomato

38 (Solanum esculentum Dunal, Solanaceae) pests in different parts of the world (Jeppson

39 et al. 1975). Although the control of this mite is done today mainly with synthetic acari-

40 cides, the evaluation of alternative means to control it is extremely desirable, especially

41 considering that tomato is one of the main horticultural crops in the world. Biological

42 control with predatory mites is a possibility to be explored.

43 The Phytoseiidae constitute the main group of predatory mites on plants. The control of

44 pest mites with phytoseiids has been successfully implemented on several crops, especially

45 the control of T. urticae with Phytoseiulus persimilis Athias-Henriot (Gerson et al. 2003).

46 Although Phytoseiulus Evans species are specific predators of Tetranychus Dufour

47 species (McMurtry and Croft 1997), the efficiency of P. persimilis in the control of

48 T. urticae on tomato is often lower than desirable (Gillespie and Quiring 1994; Drukker

49 et al. 1997). Efforts have been dedicated to obtain strains of predatory mites with better

50 performance on tomato (Drukker et al. 1997; Gerson et al. 2003), but the discovery of more

51 efficient predators is highly desirable. Promising results have been reported for a popu-

52 lation of Phytoseiulus longipes Evans found in Uruguaiana, in southern Brazil (Furtado

53 et al. 2006). This population can develop and reproduce well on Tetranychus evansi Baker

54 and Pritchard and also on T. urticae on tomato (Furtado et al. 2007). Further information

55 about the possible significant impact of P. longipes on T. evansi on tomato plants grown in

56 screen-houses was provided by Silva et al. (2010).

57 Phytoseiulus macropilis (Banks) has been reported from several tropical and subtropical

58 regions of the world (Takahashi and Chant 1993; Moraes et al. 2004), including extensive

59 regions in Brazil. Aspects of its biology have been studied by Smith and Summers (1949);

60 Prasad (1967); Takafuji and Chant (1976); Shih et al. (1979); Ali (1998). Aspects of

61 its potential as biological control agent have been studied by Ali (1998); Garcia and

62 Chiavegato (1997); Oliveira et al. (2007) and other references mentioned by Kostiainen

63 and Hoy (1996). The efficiency of this predator led several companies to mass produce and

64 commercialize it for use on different crops (Gerson et al. 2003). However, nothing has

65 been reported in relation to its possible use on tomato plants.

66 Tomato is known to contain a diversity of trichomes of variable densities, depending on

67 the variety. Trichomes are outgrows of several types found at various densities in most

68 plants; some trichomes are known to produce secretions that concentrate at their tips; those

69 are grouped as glandular trichomes, to distinguish from other that do not produce those

70 secretions, called non-glandular trichomes.

71 Trichomes have generally been reported to provide protection to plants against biotic

72 and abiotic environmental factors. Among other things, they have been reported to hamper

73 the action of phytophagous organisms, including mites. While there are references

74 attributing to those structures a deleterious effect on some predatory mites (Van Haren

75 et al. 1987; Gillespie and Quiring 1994; Nihoul 1994; Drukker et al. 1997), there are also

76 references indicating them as beneficial to other predatory mites (Karban et al. 1995; Roda

77 et al. 2000; Kreiter et al. 2003; Loughner et al. 2008).

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78 The objective of this work was to compare the performance of a Brazilian population of

79 P. macropilis on tomato with its performance on other plant species, relating the observed

80 variation with the respective types and densities of trichomes. It has been mentioned in the

81 literature that the content of glandular trichomes is released onto the leaf surface when the

82 leaf is touched or chewed by herbivores (Van Haren et al. 1987; Fridman et al. 2005). It has

83 been hypothesized (Van Haren et al. 1987) that the presence of the webbing produced by

84 T. urticae can help the predator to avoid contact with trichomes and consequently to

85 improve its performance on tomato plants; this hypothesis was also tested in the present

86 study. Functioning as a control, P. longipes was included in the study, because of its

87 reported ability to prey on T. urticae on tomato and, mostly, because we have never seen it

88 stuck to trichomes of tomato plants in our laboratory and field observations. Phytoseiulus

89 persimilis would have been a better species to be used as control. However, this species is

90 not found in Brazil, where the study was conducted.

91 Materials and methods

92 The study was conducted in a laboratory in mid-2008. The plants considered in the study

93 were ‘Carmem’ and ‘cerasiforme’ tomato varieties, nightshade (Solanum americanum

94 Mill.) and eggplant (Solanum melongena L., variety ‘Natu Nobilis’), all Solanaceae, as

95 well as Jack-bean [Canavalia ensiformis (L.) DC.], Fabaceae. ‘Carmen’ is one of the main

96 tomato varieties grown in Brazil; ‘cerasiforme’ is also widely cultivated, but it was

97 selected mainly because of the very high density of trichomes. Tomato plants were

98 obtained from plantlets acquired commercially; other plants were obtained from seeds

99 either acquired commercially (eggplant and Jack-bean) or obtained from plants occurring

100 naturally on the campus of Escola Superior de Agricultura Luiz de Queiroz, Universidade

101 de Sao Paulo (ESALQ-USP), in Piracicaba, State of Sao Paulo, Brazil (nightshade).

102 Before starting the tests, the density of trichomes on the lower surface of leaves of

103 nightshade and eggplant and leaflets of each tomato variety and Jack-bean was evaluated.

104 Trichomes were grouped as glandular and nonglandular; the terminology used to distin-

105 guish the different types of trichomes is based on Metcalfe and Chalk (1979) and Simmons

106 and Gurr (2005), the latter specifically referring to tomato. Leaves (leaflets) considered in

107 these evaluations were taken from plants of the following ages: tomato and eggplant, about

108 45 and 90 days after transplanting, respectively; nightshade and Jack-bean, about 60 and

109 30 days of germination, respectively. They were taken from the same region of the plants

110 from which similar leaves (leaflets) would be shortly afterward taken for the biological

111 tests: tomato, median leaflet of a leaf from the median region of the plant; eggplant and

112 nightshade, leaves from the median region of the plant; Jack-bean, central leaflet of the

113 most recently (totally) open leaf. Evaluations were conducted on 10 leaves (leaflets) of

114 each plant species (variety), in an area of 1.0 cm2 near the center of one of the distal

115 quarters of each leaf (leaflet). Trichomes were counted under a dissecting microscope

116 (Leica MZ8), at 50 x magnification.

117 Specimens of T. urticae were taken from a stock colony regularly maintained on Jack-

118 bean plants in a screen-house at ESALQ-USP. Specimens of P. macropilis were obtained

119 from a stock colony initiated with mites collected in 2004 from ornamentals in Holambra,

120 State of Sao Paulo, while specimens of P. longipes were obtained from a stock colony

121 initiated with mites collected in 2008 from solanaceous plants in Uruguaiana. Each

122 treatment of each test was conducted with 25 adult female predators, considering each

123 predator as a replicate. Predators were 2 days-old at the beginning of each test, and were

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124 kept in 0.5 ml plastic vials without water and food for about 15 h that preceded the

125 beginning of each test. Tests were conducted at 25 ± 2�C, 70 ± 10% RH and 14 h of

126 photophase.

127 Dispersal of predators on leaflets

128 An adaptation of the method described by Weston and Snyder (1990) was used in this

129 test. Each experimental unit consisted of a leaflet of tomato (variety ‘cerasiforme’) or of

130 Jack-bean, the latter included as a control because of its low density of trichomes. Each

131 leaflet was placed, ventral surface up, onto a 10 9 7 cm piece of styrofoam; the margins

132 of the leaflet were covered with a band of wet cotton wool and a metallic thumbtack (flat

133 head 9 mm in diameter) was inserted in the center of the leaflet. A predator was then

134 placed with a brush in the center of the thumbtack head, measuring the total distance

135 traversed by it after a period of 20 s. For this, the predator was continuously observed,

136 under a dissecting microscope, inserting a pin at each point of inflection of the path of the

137 predator’s movement, caring to reduce as much as possible interference with its normal

138 behavior. The distance was considered null when the predator did not leave the head of

139 the thumbtack. The procedure was repeated until all predators were, one by one,

140 observed.

141 Dispersal of predators on stems

142 This test was conducted with stems of the same plants mentioned for the previous test, to

143 estimate the ability of predators to move between different leaves on a tomato plant. It was

144 supposed they would have more difficulty in dispersing on stems than on leaflets, due to the

145 larger number of trichomes on the former, as observed in this study, although the

146 respective densities were not registered, and as observed by Van Haren et al. (1987) for

147 other tomato varieties. Stems of Jack-bean were used for comparison, because of their low

148 number of trichomes.

149 Each experimental unit consisted of a section of 15 cm of the median region of the main

150 stem of each of the plant species considered in the study; tomato plants were about

151 45 days-old after transplanting and Jack-bean plants, about 30 days-old after planting. The

152 proximal end of each stem was covered with a piece of masking tape and the stem was

153 placed onto a layer of cotton wool soaked in distilled water to prevent mites from escaping,

154 in a plastic tray. A Jack-bean leaflet infested 24 h before with 20 T. urticae females was

155 adjacent to the distal end of each stem, to serve as a possible attractant to the predator.

156 Soon after setting up an experimental unit, the evaluation was initiated putting a

157 predator on the exposed surface of the masking tape, determining subsequently the time

158 necessary for the predator to reach the opposite end. Also as reported for the previous test,

159 the procedure was repeated until all predators were, one by one, observed.

160 Prey consumption and oviposition

161 In the absence of webbing, each experimental unit consisted of a disk of 3.0 cm in diameter

162 of a leaf (leaflet) of a plant substrate placed onto a layer of cotton wool soaked in distilled

163 water in a Petri dish of 3.5 cm in diameter. Each unit contained 30 eggs of T. urticae (up to

164 24 h of age), transferred randomly with a brush from an infested leaf onto the rearing unit,

165 and a single predator. Each unit was kept closed with a transparent plastic film.

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166 The study was initiated transferring a predator to each experimental unit. After 24 h,

167 each predator was transferred to a new, similar unit; after another period of 24 h, the

168 process was repeated. At the end of each 24 h period, the number of eggs of T. urticae

169 consumed by each predator and the number of eggs laid by each predator were registered.

170 In the presence of webbing, the methodology adopted was similar to what was men-

171 tioned for the determination in the absence of webbing, except that prey eggs were not

172 transferred, but laid by 20 females that had been placed in each experimental unit 24 h

173 before introducing the predators. After this period, all adult females and part of the eggs

174 they laid were removed from the unit, disrupting as little as possible the webbing and

175 leaving 30 eggs in each unit. No effort was made to separate webbing from the associated

176 faeces of the prey; thus, what is subsequently mentioned as webbing in this paper actually

177 refers to both webbing and faeces.

178 Statistical analyses

179 Before comparing data of each study, tests of normality of means were performed using

180 PROC UNIVARIATE (SAS 9.1) and Shapiro–Wilk test. Data always had normal distri-

181 bution and were therefore subjected to analyses without transformations. Turkey’s

182 Studentized Range (HSD) tests were used for comparisons of more than 2 means and

183 Student’s t tests were used for comparisons of 2 means, at 5% level of significance. The

184 study of prey consumption and oviposition consisted of a factorial design in which the

185 main factors were predator species (P. macropilis, P. longipes), presence of prey webbing

186 in the experimental unit (absent, present) and substrate (experimental units prepared with

187 leaves or leaflets of ‘Carmem’ and ‘cerasiforme’ tomato varieties, eggplant, nightshade and

188 Jack-bean). Statistical comparisons of means were done only for the main factors and

189 interactions of main factors for which significant differences were detected in the factorial

190 analyses.

191 Results

192 The number of glandular trichomes was significantly higher in the ‘cerasiforme’ tomato

193 variety, followed by the ‘Carmem’ tomato variety, Jack-bean and nightshade (Table 1).

194 Trichomes of eggplant were not evaluated [Leite et al. (2003) did not find glandular

195 trichomes on leaves of the ‘Natu Nobilis’ eggplant variety]. About 94 and 70% of the

196 glandular trichomes of ‘cerasiforme’ and ‘Carmem’ varieties, respectively, were classified

197 as type VI; the remaining glandular trichomes of those varieties were classified as type I.

198 All glandular trichomes of nightshade and Jack-bean were simple, similar to type I of

199 tomato, but shorter.

200 Nonglandular trichomes were much more numerous in the ‘Carmem’ tomato variety,

201 followed by the ‘cerasiforme’ tomato variety, Jack-bean and nightshade. Considering the

202 data of Leite et al. (2003), nonglandular trichomes seem still much more numerous in the

203 ‘Natu Nobilis’ eggplant variety. All nonglandular trichomes of both tomato varieties were

204 simple (unbranched). About 77 and 92% of these trichomes were classified as type V in

205 ‘Carmem’ and ‘cerasiforme’ varieties, respectively; the remaining nonglandular trichomes

206 of those varieties were classified as type III. All nonglandular trichomes of nightshade and

207 Jack-bean were simple. In the ‘Natu Nobilis’ eggplant variety, both simple and ramified

208 (stellate) nonglandular trichomes are found, the latter corresponding to about 89% of all

209 trichomes (Leite et al. 2003).

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210 Dispersal of predators

211 No significant differences were found between P. macropilis and P. longipes in relation to

212 the distances traversed on the leaflets of either the ‘cerasiforme’ tomato variety or Jack-

213 bean (Table 2). Both predator species traversed distances significantly shorter on leaflets of

214 the former.

215 Similarly to what was observed for leaflets, no significant differences were observed

216 between P. macropilis and P. longipes in relation to the period of time required to reach the

217 distal end of the stem of those plants (Table 3). Also, both predator species required

218 significantly much longer to reach the distal end of the stem of the ‘cerasiforme’ tomato

219 variety.

220 Prey consumption and oviposition

221 The factorial analyses showed no significant differences between predator species for

222 either prey consumption or predator oviposition (Table 4). However, significant differ-

223 ences were observed for the other main factors: presence of webbing and substrates. In

224 relation to interactions, significant differences were only observed for the interaction

225 between presence of webbing in the experimental unit and substrate.

226 Given the absence of significant differences between predator species, subsequent

227 analyses were performed with the pooled data obtained for both P. macropilis and

228 P. longipes. Both prey consumption and predator oviposition rates were significantly lower

Table 1 Number of trichomes per cm2 (±SE) on leaves (leaflets) of the plant species considered in this

study

Substrate Types of trichomesa

Glandulars Nonglandulars

S. esculentum var. ‘Carmem’ 133.6 ± 3.2 b 302.0 ± 2.5 a

S. esculentum var. ‘cerasiforme’ 204.4 ± 3.3 a 207.7 ± 3.4 b

C. ensiformis 16.8 ± 0.8 c 137.5 ± 4.2 c

S. americanum 11.6 ± 0.9 c 12.8 ± 1.3 d

S. melongena var. ‘Natu Nobilis’b 0 ca. 570

a Means followed by the same letters in the column are not statistically different (Tukey, P\ 0.05)b Approximation of the average for the lower surface of median leaves, calculated from data given by Leite

et al. (2003)

Table 2 Distances traversed in 20 s (cm ± SE) by Phytoseiulus macropilis and Phytoseiulus longipes on a

leaflet of the ‘cerasiforme’ tomato variety (Solanum esculentum) or of Cannavalia ensiformis

Predator Planta

Tomato var. ‘cerasiforme’ C. ensiformis

P. macropilis 2.4 ± 0.2 a B 4.3 ± 0.2 a A

P. longipes 2.6 ± 0.1 a B 4.9 ± 0.2 a A

a Means followed by the same small case letters in the column or by the same capital letters on the row are

not statistically different (t tests, P\ 0.05)

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229 in the ‘cerasiforme’ tomato variety than in other substrates, while in most cases the latter

230 did not significantly differ between themselves (Table 5). The only exception was eggplant

231 in the absence of prey webbing (hence the significance of the interaction between prey

232 webbing and substrates); in this case, prey consumption and oviposition did not differ

233 significantly from what was observed for the ‘cerasiforme’ tomato variety.

234 In all substrates, prey consumption and predator oviposition was significantly higher on

235 leaves (leaflets) where prey webbing was present.

236 Discussion

237 The performance of P. macropilis was similar to that of P. longipes in relation to the ability

238 to disperse, to prey on T. urticae and to oviposit, independently of the type and density of

239 trichomes and the presence or absence of prey webbing.

240 The less adequate performance of both predators in the ‘cerasiforme’ variety of tomato,

241 as indicated by the lower ability to walk on each leaflet, to disperse from leaf to leaf by

Table 3 Time (seconds ± SE) required by Phytoseiulus macropilis and Phytoseiulus longipes to traverse a

distance of 15 cm of a stem of ‘cerasiforme’ tomato variety (Solanum esculentum) or of Cannavalia

ensiformis

Predador Planta

Tomato var. ‘cerasiforme’ C. ensiformis

P. macropilis 123.6 ± 8.0 b A 12.1 ± 0.2 b B

P. longipes 117.5 ± 6.2 b A 11.8 ± 0.2 b B

a Means followed by the same small case letters in the column or by the same capital letters on the row are

not statistically different (t tests, P\ 0.05)

Table 4 Factorial analyses on

the daily consumption of eggs

of Tetranychus urticae and daily

oviposition rate of predators

Main factors: Predators

(Phytoseiulus macropilis,

Phytoseiulus longipes); prey

webbing (present, absent);

substrate [tomato (Solanum

esculentum) var. ‘Carmem’

and ‘cerasiforme’, Canavalia

ensiformes, Solanum melongena,

Solanum americanum]

Factor df F P[F

Prey consumption

Substrate 4 46.76 \.0001

Predator 1 0.40 0.5275

Webbing 1 192.17 \.0001

Substrate 9 predator 4 0.73 0.5739

Substrate 9 webbing 4 8.65 \.0001

Predator 9 webbing 1 0.01 0.9394

Substrate 9 predator 9 webbing 4 0.39 0.8150

Oviposition

Substrate 4 8.98 \.0001

Predator 1 0.03 0.8699

Webbing 1 257.38 \.0001

Substrate 9 predator 4 1.08 0.3681

Substrate 9 webbing 4 2.41 0.0485

Predator 9 webbing 1 0.05 0.8315

Substrate 9 predator 9 webbing 4 0.46 0.7645

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242 walking on the stem as well as by the lower prey consumption and oviposition rates, was

243 related to the much higher density of glandular trichomes on this plant. However, even

244 under that extreme condition, the predators were not observed to become stuck to the

245 trichomes and, despite more slowly, it did disperse along the stem. Thus, it is assumed that

246 the predators could also move from leaf to leaf of the ‘Carmem’ variety, without getting

247 stuck In addition, the glandular trichomes in the ‘Carmem’ tomato variety did not sig-

248 nificantly interfere with prey consumption and oviposition rates of either predator species,

249 as the determined levels of those parameters were comparable to those obtained on

250 nightshade and Jack-bean, which have very few glandular trichomes. Trichomes of egg-

251 plant, though nonglandular, interfered significantly with the performance of both predator

252 species, as indicated by the much lower prey consumption and oviposition rates than

253 verified for nightshade and Jack-bean.

254 The lower prey consumption and oviposition rate of P. macropilis and P. longipes on

255 leaflets of the ‘cerasiforme’ tomato variety indicates the presence of one or more unfa-

256 vorable factor(s) to the predator on the surface of those leaflets, which was (were) only

257 partially suppressed by the presence of prey webbing, as indicated by the fact that both

258 prey consumption and predator oviposition rates were significantly higher when webbing

259 was present than when it was absent on the same substrate, but still lower than for

260 nightshade and Jack-bean substrates with webbing. Conversely, the presence of webbing

261 was sufficient to eliminate the disturbing effect of the trichomes of eggplant, as indicated

262 by the fact that when webbing was present, prey consumption and oviposition rates of the

263 predators were not statistically different from the rates observed on nightshade and Jack-

264 beansubstrates with webbing.

265 The results of the present study indicated that trichomes may interfere with the pre-

266 dators by means other than entrapment in exudates of glandular trichomes. None of the

267 predators used in this study was ever seen stuck to trichomes. This observation contrasts

268 with the results obtained by Van Haren et al. (1987) for P. persimilis; these authors

269 observed the entrapment of 35 to 61% of the specimens of that species on stems of plants

270 of the ‘Turbo’ tomato variety. Nihoul (1994) also reported mortality of P. persimilis on

271 leaves and stems of tomato plants, due to the entrapment of mites of this species in

Table 5 Daily consumption

of eggs of Tetranychus urticae

and daily oviposition rate (±SE)

of Phytoseiulus macropilis and

Phytoseiulus longipes, in the

absence and in the presence

of prey webbing on different

substrates

a Number of eggs consumedb Number of eggs laid. Within

each parameter, averages

followed by the same small case

letters in the column or by the

same capital letter in the row are

not statistically different (Tukey,

P\ 0.05; t tests, P\ 0.05)

Substrate Webbing

Absent Present

Prey consumptiona

Tomato var. ‘Carmem’ 17.2 ± 0.4 a B 20.6 ± 0.3 a A

Tomato var. ‘cerasiforme’ 10.5 ± 0.5 b B 15.8 ± 0.4 b A

C. ensiformis 16.7 ± 0.5 a B 19.5 ± 0.4 a A

S. melongena 11.3 ± 0.3 b B 19.3 ± 0.4 a A

S. americanum 16.9 ± 0.3 a B 20.2 ± 0.4 a A

Ovipositionb

Tomato var. ‘Carmem’ 1.6 ± 0.1 a B 2.3 ± 0.1 a A

Tomato var. ‘cerasiforme’ 1.0 ± 0.1 b B 2.0 ± 0.1 b A

C. ensiformis 1.6 ± 0.1 a B 2.4 ± 0.1 a A

S. melongena 1.2 ± 0.1 b B 2.3 ± 0.1 a A

S. americanum 1.6 ± 0.1 a B 2.4 ± 0.1 a A

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272 exudates of glandular trichomes. The reasons for the differences between the results of the

273 present study and of the studies reported by those authors are not known. One of the

274 reasons could be the difference in size and the related ‘‘vigor’’ of the predators involved.

275 However, measurements taken of samples of the species considered in the present study

276 and of specimens of P. persimilis available in reference collection of ESALQ-USP did not

277 show any major differences between them. Another possible reason could be the differ-

278 ences between the plants used in this and in the previous studies: ‘Turbo’ variety in the

279 study of Van Haren et al. (1987), and ‘2209 De Ruiter’ variety in the study of Nihoul

280 (1994); in the latter, the number of glandular trichomes was most often higher than

281 determined in the present study. It has been determined that even within the same variety

282 the number of trichomes can be influenced by the growing conditions of the plants

283 (Gianfagna et al. 1992; Nihoul 1993).

284 It seems possible that in the ‘cerasiforme’ variety one of the factors hampering the

285 performance of the predators was chemical, given the fact that in the presence of webbing

286 the performance of both predators remained less adequate than observed on nightshade and

287 Jack-bean. There are at least two possible explanations as to how the putative chemical

288 effect could have occurred. First, it is possible that when removing females and surplus

289 eggs from the rearing units, before initiating the test, some of the glandular trichomes were

290 touched with the brush, releasing their secretion. Alternatively, it is possible that the

291 webbing produced by T. urticae was not enough to cover the total surface of the experi-

292 mental unit, allowing predators to touch some of those trichomes, causing the release of

293 their secretion. The main component of the exudates of tomato glandular trichomes

294 (2-tridecanone) is only released to the environment when the top cuticle of the glands is

295 broken (Van Haren et al. 1987). That component is known to be toxic to insects, but its

296 toxicity or repellence to Phytoseiulus species seems not to have been reported yet (Nihoul

297 1994). Its repellence to T. urticae is already known (Cantelo et al. 1974; Snyder and Carter

298 1984).

299 The hampering effect of nonglandular trichomes of eggplant might be only physical, as

300 suggested by the fact that prey consumption and consequent oviposition rates were similar

301 to what was observed on nightshade and Jack-bean, when webbing was available. This

302 phenomenon would be expected, given that nonglandular trichomes are not expected to

303 release chemicals, interfering with tiny arthropods, when too numerous, mostly by phys-

304 ically disturbing their movements. Krips et al. (1999) also reported lower speed of dis-

305 persal of P. persimilis on cultivars ‘Sirtaki’ and ‘Bourgone’ than on cultivar ‘Bianca’ of

306 Gerbera jamesonii Adlam, the latter of which contains the lowest density of (nonglandular)

307 trichomes.

308 However, while webbing could have a physically favorable effect on P. macropilis and

309 P. longipes by possibly reducing their contact with trichomes, its effect in favoring prey

310 consumption and the consequent predator oviposition rates was clearly shown by the fact

311 that these were much higher on all substrates, including those with few trichomes, when

312 webbing was present. Webbing in itself may not provide chemical cues to stimulate

313 predation and oviposition. Sabelis et al. (1984) could not find evidences for the presence of

314 a kairomone to attract P. persimilis in the webbing produced by T. urticae. However, they

315 did find evidences of the presence of a kairomone in the faeces of T. urticae. Because in the

316 present study faeces of T. urticae were not separated from the webbing, it seems con-

317 ceivable that predators could have been stimulated to oviposit, at least partially because of

318 the faeces. In trying to elucidate the actual effect of spider mite webbing on P. persimilis,

319 Roda et al. (2001) showed that it might favor survival and fitness of P. persimilis in ways

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320 not related to the indication of the presence of prey, but rather the adequate physical

321 structure it provides, suitable as oviposition site.

322 Interestingly, oviposition of the predators occurred mostly near the midrib of leaves of

323 nightshade and leaflets of Jack-bean, regions where glandular trichomes were concentrated.

324 This could suggest that, up to a certain density, the net effect of those structures on

325 predators may be positive. Examples of beneficial effects of nonglandular trichomes on

326 phytoseiids have been summarized by several authors (Kreiter et al. 2003; Loughner et al.

327 2008). However, it is probable that the preference of P. macropilis and P. longipes for

328 ovipositing next to the midrib of nightshade and Jack-bean refers at least in part to the

329 protection the midrib itself provides to eggs; even on glabrous leaves, it is quite common

330 for phytoseiids to oviposit next to the midrib. Alternatively, the preference for oviposition

331 at this site could be due to the prevailing conditions of temperature and humidity; Cook

332 and Dixon (1964) observed lower temperature near the midrib of tomato leaflets as

333 compared to other sites, what was assumed to be associated with a higher transpiration rate

334 in the former.

335 The results of this study suggest that P. macropilis could represent an option for the

336 biological control of T. urticae on tomato, especially in places where the former occurs and

337 the species commercially used for that purpose, P. persimilis, is not available. It is not

338 possible at this stage to compare the performance of those predators; although P. mac-

339 ropilis seems to be less prone to become stuck to trichome exsudates, laboratory works

340 indicate that the oviposition rate of P. persimilis is considerably higher (Gillespie et al.,

341 1994; Castagnoli et al. 1998) than what was determined in this study for P. macropilis.

342 In addition, the ability of P. macropilis to feed and reproduce on T. urticae was evaluated

343 only for a short period. Complementary studies should be conducted with this predator,

344 along the phenological cycle of tomato.

345 Acknowledgments To M. Bellini, PROMIP (Brazil), for helping in obtaining the mites used in this study.346 To CNPq (Brazil), for the financial support to the first and the second authors. To B. Apezzato-da-Gloria, for347 the help in the quantification of trichomes.

348

349 References

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