Insecticidal effect of Dittrichia viscosa lyophilized epicuticular material against four major stored-product beetle species on wheat

Journal Pre-proof Insecticidal effect of Dittrichia viscosa lyophilized epicuticular material against four major stored-product beetle species on wheat E. Lampiri, P. Agrafioti, E. Levizou, C.G. Athanassiou PII:

S0261-2194(20)30028-4

DOI:

https://doi.org/10.1016/j.cropro.2020.105095

Reference:

JCRP 105095

To appear in:

Crop Protection

Received Date: 1 December 2019 Revised Date:

27 January 2020

Accepted Date: 29 January 2020

Please cite this article as: Lampiri, E., Agrafioti, P., Levizou, E., Athanassiou, C.G., Insecticidal effect of Dittrichia viscosa lyophilized epicuticular material against four major stored-product beetle species on wheat, Crop Protection (2020), doi: https://doi.org/10.1016/j.cropro.2020.105095. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2020 Published by Elsevier Ltd.

1

Crop Protection

Prof. Christos G. Athanassiou

2

Laboratory of Entomology and Agricultural Zoology

3

University of Thessaly

4

Phytokou str., 38446, Nea Ionia Magnessia, Greece

5

Phone: +30-2421093195

6

FAX: +30-2421093178

7

e-mail: [email protected]

8

9

Insecticidal Effect of Dittrichia viscosa lyophilized epicuticular material against four

10

major stored-product beetle species on wheat

11

E. Lampiri a,b, P. Agrafioti a, E. Levizou b, C.G. Athanassiou a*

12

13

a

14

Production and Rural Environment, University of Thessaly, Phytokou str., Nea Ionia,

15

Magnesia, 38446, Greece

16

b

17

Environment, University of Thessaly, Phytokou str., Nea Ionia, Magnesia, 38446, Greece

18

(* corresponding author, e-mail address: [email protected])

Laboratory of Entomology and Agricultural Zoology, Department of Agriculture, Crop

Laboratory of Weed Science, Department of Agriculture, Crop Production and Rural

19

20

1

21

ABSTRACT

22

We examined the insecticidal effect of lyophilized epicuticular material of the ruderal species

23

Dittrichia viscosa in four major stored-product beetle species. Furthermore, the potential of

24

this material in progeny production suppression was also evaluated. The water-soluble extract

25

was derived from plants that had been harvested in September 2016, through freeze-drying, in

26

order to create a fine powder formulation. In our bioassays, the powder was applied in four

27

doses on wheat: 0 (control), 1000, 3000 and 5000 ppm and mortality of the exposed

28

individuals was measured after 1, 3, 7, 14 and 21 days of exposure, while progeny production

29

capacity was recorded 65 days later. Among the species tested, Oryzaephilus surinamensis

30

was found to be the most susceptible, followed by Tribolium confusum and Sitophilus oryzae,

31

while Rhyzopertha dominica was not practically affected. Progeny production was

32

particularly reduced for all species relative to the controls. Indicatively, for O. surinamensis,

33

at the highest dose rate, there were only 0.2 adults per vial, while the respective figures for the

34

control exceeded 40 adults per vial. To our knowledge, this study is the first that examined the

35

insecticidal effect of epicuticular material of D. viscosa for the control of stored-grain insect

36

species. Additional experimentation is required to indicate the rationale of using this natural

37

resource-based material under a non-chemical control strategy at the post-harvest stages of

38

agricultural commodities.

39

40

Keywords: Dittrichia viscosa, lyophilized epicuticular material, botanicals, post-harvest

41

control, stored-product insects

42

43

2

44

1. Introduction

45

The most common method that is currently in use for the control of stored-product pests is

46

chemical insecticides, which are mainly separated into two major categories: contact

47

insecticides and fumigants, such as grain protectants and phosphine (Athanassiou and Arthur,

48

2018; Daglish et al., 2018). Despite the positive effects that conventional insecticides can

49

provide against stored-product pests, over the past few decades, there are increasing concerns

50

regarding their use for health and environmental reasons, which has led to the withdrawal of

51

many active ingredients (Dubey et al., 2010; Nayak and Daglish, 2018; Stejskal et al., 2018).

52

Furthermore, a major issue resulting from the continuous use of chemical control is the

53

development of resistance of key stored-product insect species to many of the currently used

54

conventional insecticides. For instance, the red flour beetle, Tribolium castaneum (Herbst)

55

(Coleoptera: Tenebrionidae) (Jagadeesan et al., 2012; 2013) and the lesser grain borer,

56

Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) (Schlipalius et al., 2002; Kaur et al.,

57

2013), were found to be resistant to phosphine. Hence, it is essential to evaluate alternatives

58

that are viable and effective, and at the same time have reduced environmental impact and

59

human health risks.

60

Numerous papers have suggested insecticides of botanical origin for use at the post-harvest

61

stages of agricultural commodities (Prakash and Rao, 1997; Weaver and Subramanyam, 2000;

62

Athanassiou et al., 2014). It is generally considered that plant derivatives fulfill the above

63

requirements, since, in majority, are non-toxic to mammals and have natural origin. The most

64

well-studied plant for this purpose is by far the neem tree, Azadirachta indica A. Juss.

65

(Sapindales: Meliaceae), and its main active ingredient azadirachtin which has been evaluated

66

with success against a wide range of stored-product insect species (Weaver and

67

Subramanyam, 2000; Athanassiou et al., 2005). Nevertheless, azadirachtin requires elevated

68

dose rates to act, which are far higher than those of the currently used conventional 3

69

insecticides (Athanassiou et al., 2005). Other well-studied plants for stored-product protection

70

are the species of the genera Brassica, Citrus, Cymbopogon, Mentha and Rosmarinus, which

71

have been effective for both stored-product beetles and moths (Prakash and Rao, 1997;

72

Weaver and Subramanyam, 2000). Indicatively, derivatives from Rosmarinus officinalis L.

73

(Lamiales: Lamiaceae), Mentha microphylla C. Kock (Lamiales: Lamiaceae) and Mentha

74

viridis L. (Lamiales: Lamiaceae) were able to control various species, such as Sitophilus spp.

75

(Coleoptera: Curculionidae), Callosobruchus spp. (Coleoptera: Bruchidae), the cigarette

76

beetle, Lasioderma serricorne (F.) (Coleoptera: Anobiidae), the bean weevil, Acanthoscelides

77

obtectus (Say) (Coleoptera: Bruchidae) (Huang et al., 2000), T. castaneum (Lee et al., 2002),

78

the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae)

79

and the Mediterranean flour moth, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) (Tunc

80

et al., 2000). In this context, one of the most important families that have been evaluated

81

towards this direction is Asteraceae, with several plants and plant substances to pose a

82

noticeable insecticidal activity (Pascual-Villalobos and Robledo, 1998; Torres et al., 2003;

83

Pavela, 2004; Boussaada et al., 2008). For instance, Anacyclus clavatus Pers. (Asterales:

84

Asteraceae) and Asteriscus maritimus (L.) Less. (Asterales: Asteraceae) were toxic to T.

85

castaneum (Pascual-Villalobos and Robledo, 1998). However, there is still inadequate

86

information for one of the Asteraceae species, the false yellowhead, Dittrichia viscosa (L.)

87

Greuter, despite the fact that there are some initial works that show a potential insecticidal

88

value (Allal-Benfekih et al., 2011; Mamoci et al., 2012; Grauso et al., 2019; Rotundo et al.,

89

2019). This kind of plant is an evergreen, perennial shrub with rich sticky foliage and intense

90

odor, widely known for its medicinal use (Parolin et al., 2014). The application of this plant,

91

either as an extract from plant tissues or as an essential oil, has shown antifungal activity

92

against Fusarium moniliforme J. Sheld. (Hypocreales: Nectriaceae), Sclerotinia sclerotiorum

93

(Lib.) de Bary (Helotiales: Sclerotiniaceae), Rhizoctonia solani J.G. Kuhn (Cantharellales:

4

94

Ceratobasidiaceae) and Phytophthora capsici Leonian (Peronosporales: Peronosporaceae)

95

(Yegen et al., 1992). Moreover, D. viscosa has a repulsive or even an acaricidal action against

96

the carmine spider mite, Tetranychus cinnabarinus (Boisduval) (Prostigmata: Tetranychidae)

97

(Topakci et al., 2005), and is effective for the control of certain plant parasitic nematodes,

98

such as Meloidogyne javanica (Treub) Chitwood (Tylenchida: Meloidogyne) (Oka et al.,

99

2001). In addition, its application as a plant extract to larvae of Tuta absoluta (Meyrick)

100

(Lepidoptera: Gelenchidae) (Allal-Benfekih et al., 2011) and to Myzus persicae (Sulzer)

101

(Hemiptera: Aphididae) (Mamoci et al., 2012) have shown positive results.

102

Most of the published works are focused on D. viscosa shoot (Qasem et al., 1995) and extracts

103

from different aerial parts of the plant or applications of its essential oil (Rotundo et al., 2019;

104

Grauso et al., 2019). Specifically, hexane and ethanolic extracts of D. viscosa have shown

105

antifeedant activity against M. persicae, Rhopalosiphum padi (L.) (Hemiptera: Aphididae)

106

and Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) (Mamoci et al., 2012; Grauso

107

et al., 2019), while high constant toxicity was found when tested against the granary weevil,

108

Sitophilus granarius (L.) (Coleoptera: Curculionidae) (Rotundo et al., 2019). However, there

109

is another plant part that has never been evaluated towards this direction: its epicuticular

110

material. The epicuticular material of D. viscosa is a water-soluble mixture of flavonoids and

111

sesquiterpenes, which is attached to the waxy surface of the cuticle (Stavrianakou et al.,

112

2004). Many eco-physiological roles have been ascribed to this exudate, such as the

113

prevention of excessive water loss via transpiration and the increased protection against

114

ultraviolet radiation. Moreover, the allelopathic potential of the epicuticular material is well-

115

documented, which successfully inhibits the germination of the neighboring plants seeds

116

and/or interferes with their radicle growth and seedling development, whilst is not toxic for its

117

own seeds and seedlings (Stephanou and Manetas, 1997; Levizou et al., 2002).

5

118

Considering the lack of information regarding the epicuticular material of D. viscosa, we have

119

carried out laboratory bioassays to examine its insecticidal effect for the control of four major

120

stored-product beetle species, two primary colonizers, R. dominica and the rice weevil,

121

Sitophilus oryzae (L.) (Coleoptera: Curculionidae) and two secondary colonizers , T.

122

confusum and the saw-toothed grain beetle , Oryzaephilus surinamensis (L.) (Coleoptera:

123

Silvanidae). Apart from parental mortality, suppression on progeny production capacity was

124

also examined.

125 126

2. Materials and Methods

127

2.1 Test Insects

128

All species were reared at the Laboratory of Entomology and Agricultural Zoology,

129

Department of Agriculture, Crop Production and Rural Environment, University of Thessaly,

130

at 25 oC, 65% relative humidity (r.h.) and continuous darkness. Rhyzopertha dominica and S.

131

oryzae were reared on whole wheat kernels, T. confusum on wheat flour and O. surinamensis

132

on oat flakes. Adult beetles, <1 month-old, were used in the tests. All rearings have been kept

133

in laboratory conditions for more than 20 years.

134

2.2 Plant epicuticular material

135

Aerial parts of D. viscosa were collected during September 2016 from rural areas of Volos

136

(Thessaly, Central Greece) and soaked in water for 3 hours. Then, the aqueous solution

137

(containing the epicuticular material) was collected, centrifuged and its absorbance at 290 nm

138

(OD290) was measured using a spectrophotometer (UV-1900, Shimatzu, Japan) after the

139

appropriate dilutions. This procedure resulted to a three-fold more concentrated extract

140

(OD290 =25) in comparison with the concentration naturally found on leaves. Then, freeze-

141

drying was followed in a vacuum concentrator (Azbil Telstar Technologies, S. L. U.) at a

6

142

temperature of -50 ° C and pressure of 0.02 mBar. Finally, the epicuticular material, formed

143

as a powder, was collected, weighed and stored at 4° C until used.

144

2.3 Commodities

145

Untreated, clean and uninfected soft wheat was used in the tests. Before the experiments, the

146

wheat was kept in cold storage (-20 o C) for at least two weeks to eliminate possible insect

147

infestation.

148

2.4 Bioassays

149

Lots of 500 g of wheat were placed in glass jars of 1000 ml in capacity and treated with the

150

powder formulation in 4 doses rates: 0 (control), 1000, 3000 and 5000 ppm. An additional

151

series of lots, in which the formulation was not applied, was used as a control. The jars were

152

then shaken for 5 minutes to ensure that the formulation was equally distributed throughout

153

the wheat mass. For the tests, we used plastic cylindrical vials (3 cm in diameter, 8 cm high,

154

Rotilabo Sample tins Snap on lid, Carl Roth, Germany), with the top one quarter of the inside

155

“neck” covered with Fluon (polytetrafluoroethylene; Northern Products, Woonsocket, USA)

156

to avoid insects’ escape. Each vial contained 20 g of treated or untreated wheat, and then 20

157

adults were placed into each vial, using different vials for each insect species and dose. All

158

vials were maintained in incubators set at at 26 °C, 55% r. h. and continuous darkness. The

159

mortality of the exposed beetles was recorded after 1, 3, 7, 14 and 21 days. For each species-

160

dose combination there were three vials, while the entire procedure was repeated three times,

161

i.e. there were three replicates with three sub-replicates, and new lots of treated and untreated

162

grains each time (3 jars X 3 vials each= 9 vials for each combination). At the end of this

163

interval, all adults (dead and alive) were removed from the vials and the vials remained in the

164

same conditions for an additional period of 65 days. Then, the vials were opened for the last

165

time and the number of progeny was recorded.

166

2.5 Data analysis

7

167

Control mortality was generally low for all species, and did not exceed 10 % in the vast

168

majority of the cases. Since the same vials were examined for mortality at the different

169

exposure intervals (1-21 days), mortality data were analyzed by using a Multivariate Analysis

170

of Variance (MANOVA-Fit with Wilk's Lambda) with concentration and exposure time as the

171

main effects, by using the JPM 8 software (SAS Institute Inc., Cary, North Carolina, USA). In

172

the case of progeny production, the data were analyzed by using an Analysis of Variance

173

(ANOVA), with dose rate as main effect. Means were separated by using the Tukey-Kramer

174

Honestly Significant Difference (HSD) test, at a level of 0.05 (Sokal and Rohlf, 1995).

175 176

3. Results

177

3.1 Adult mortality

178

Regarding parental mortality, MANOVA indicated that both dose and its interaction with

179

exposure were significant for adults of O. surinamensis, T. confusum and S. oryzae (Table 1).

180

In contrast, only exposure was found to be significant in the case of R.dominica (Table 1). For

181

this species, regardless of the dose rate, adult mortality did not differ significantly from the

182

control on any of the exposure intervals, and did not exceed 12% (Table 2). Similarly,

183

mortality of S. oryzae adults was extremely low, and did not exceed 33 % for any of the dose-

184

exposure combinations tested (Table 3). However, the application of the formulation resulted,

185

at exposures that were longer than 1 day, in higher mortality levels than those in the control

186

vials, for many of the combinations tested. Furthermore, at exposures that were longer than 3

187

days, there were significant differences among treatments for adult mortality of T. confusum

188

(Table 4). Still, despite any differences, mortality was low and did not exceed 37 %. Finally,

189

O. surinamensis was proved to be the most susceptible species from the ones tested, as

190

mortality reached 87 % after 21 days of exposure at the highest dose rate (Table 5). In fact,

191

with the exception of 1 day, for all other exposures there were significant differences among

8

192

treatments. At the 7, 14 and 21 days of exposure, significantly more adults were dead in vials

193

containing 3000 and 5000 ppm of the D. viscosa formulation, as compared with the respective

194

figures of control and 1000 ppm. Moreover, at these exposures, 5000 ppm gave significantly

195

higher mortality levels than those in 3000 ppm.

196

3.2 Progeny production

197

Regarding progeny production counts, dose had a significant effect for all species, with the

198

exception of S. oryzae (Table 6). For R. dominica, the increase of dose decreased progeny

199

production, but only 3000 and 5000 ppm gave fewer offspring than that in the control (Table

200

7). Nevertheless, even in these doses, progeny production was high, and exceeded 21 adults

201

per vial. In contrast, for S. oryzae adults, the application of the formulation on wheat did not

202

significantly affect progeny production capacity, despite the fact that fewer adults were

203

recorded at 5000 ppm. For both T. confusum and O. surinamensis, the application of the D.

204

viscosa formulation at 3000 and 5000 ppm significantly suppressed progeny production in

205

comparison with the control vials. Hence, there were less than 1 adult per vial at 3000 and

206

5000 ppm and at 5000 ppm for T. confusum and O. surinamensis, respectively (Table 7).

207 208

4. Discussion

209

To our knowledge, this is the first work that has examined the insecticidal efficacy of the

210

lyophilized epicuticular material of D. viscosa. This species has been the subject of many

211

studies concerning the antimicrobial effects of plant extracts as well as the allelopathic

212

potential of its exudates, but there is still inadequate information regarding its insecticidal

213

effects on stored product insects. Chromatographic separation of the epicuticular material of

214

D. viscosa, carried out by Daniewski et al. (1986), isolated illicic acid, 2α-hydroxyisocostic

215

acid and methyl ester of illicic acid. In that study, the authors tested the action of these

216

components as food deterrents against adults and larvae of T. confusum and the khapra beetle,

9

217

Trogoderma granarium Everts (Coleoptera: Dermestideae), as well as adults of S. granarius.

218

The results shown that the 2α-hydroxyisocostic acid and methyl ester of illicic acid were

219

highly effective as food deterrents for S. granarius, whereas illicic acid and methyl ester of

220

illicic acid were highly effective for T. confusum larvae. The least susceptible life stage

221

examined was T. granarium larvae, as they were less affected by all components of the

222

epicuticular material. The above research showed that the component with the highest

223

suppressive effect was 2α-hydroxyisocostic acid, followed by methyl ester of illicic acid and

224

illicic acid. The above study is the only work that has examined components of the

225

epicuticular material of D. viscosa for the control of stored-product insects; still, that approach

226

was related with behavioral aspects, i.e. antifeedant effects, and not the evaluation of this

227

material as an insecticide/grain protectant.

228

Our results clearly indicate that the efficacy of this material varies among the species tested,

229

and it is highly related with the dose rate. Practically, this material has no effect in the case of

230

R. dominica, despite the fact that there was some suppression in progeny production.

231

Similarly, mortality and progeny production control was rather low for S. oryzae. As both

232

species are primary colonizers, and their immature development occurs within the grain

233

kernel (Athanassiou and Arthur, 2018), their larvae may remain unaffected by substances that

234

have been applied in the external kernel part. Still, the progeny production suppression for R.

235

dominica was higher than that of S. oryzae, despite that the reverse was true for mortality.

236

This could be attributed to the fact that, although immature development for R. dominica

237

occurs within the kernel, oviposition occurs at the external kernel part, and newly-hatched

238

larvae have to crawl to enter the kernel (Mayhew and Phillips, 1994; Batta, 2005). In contrast,

239

for S. oryzae, females oviposit directly inside the kernels where egg hatching takes place

240

(Haines, 1991; Rita Devi et al., 2017). For diatomaceous earths (DEs), Arthur and Throne

241

(2003), showed that there was no effect to larvae of S. oryzae when DEs were applied on

10

242

already infested grains. Similarly, probably for the same reason, R. dominica is considered as

243

one of the least susceptible species to DEs (Korunic, 1998). There are several paradigms on

244

which primary colonizers are more tolerant than secondary colonizers, i.e. species on which

245

immature development occurs outside of the kernel, to botanicals/plant extracts. For instance,

246

Hernandez- Lambrano et al. (2015) confirmed that O. surinamensis was more susceptible than

247

the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) when exposed

248

to essential oils produced by three plants of the genus Cymbopogon. In addition, the work of

249

Tapondjou et al. (2005) showed that T. confusum was more susceptible than S. zeamais when

250

exposed to cymol, the major component of essential oil of Eucalyptus saligna Sm. (Myrtales:

251

Myrtaceae). However, given that the mode of action of D. viscosa is not well-known,

252

additional experimental work is required to illustrate the basis of these differences among

253

species that share the same habitat in the stored-grain ecosystem.

254

In contrast with the primary colonizers, O. surinamensis and T. confusum were found to be

255

more susceptible to the material tested, as progeny production was significantly reduced in the

256

treated grains, in comparison with the respective figures for the untreated grains, regardless of

257

the level of parental mortality. From the species tested, O. surinamensis was found to be by

258

far the most susceptible, as parental mortality was high, even at short exposure intervals.

259

Moreover, for this species, parental mortality was notably increased with the increase of the

260

exposure interval, and would probably reach 100 % at longer exposures. At the same time,

261

progeny suppression was high at 3000 and 5000 ppm, which may be considered as a direct

262

consequence of parental mortality. Given that the immatures of the secondary colonizers

263

develop and move freely at the external kernel part, it is generally expected that their contact

264

with the D. viscosa material around the grain kernels would be substantially increased, as

265

compared with the immatures of the primary colonizers. Conversely, progeny production

266

suppression of T. confusum may not be linked with parental morality, as this was much lower

11

267

than that of O. surinamensis. It is well established that T. confusum cannot develop easily in

268

sound kernels (Aitken, 1975), which stands in accordance with our findings, as progeny

269

production in the untreated wheat was extremely low. Still, progeny production was

270

significantly suppressed in the treated grains, which consists an additional indication of the

271

adverse effect of the lyophilized epicuticular material of D. viscosa on immature development

272

and longevity.

273

The dose rates that have been used here can be considered as high, at least in comparison with

274

traditional grain protectants that are usually applied at concentrations that do not exceed 10

275

ppm (Arthur, 1996; Weaver and Subramanyam, 2000; Athanassiou et al., 2014). Moreover,

276

the lyophilized epicuticular material of D. viscosa, at least at the conditions tested here, can be

277

considered as slow-acting towards its insecticidal efficacy, as compared with the majority of

278

the conventional grain protectants. Nevertheless, there are some registered insecticides that

279

are applied directly on grains at dose rates that are comparable with the ones tested here, such

280

as DEs and zeolites (Rojht et al., 2010; Rumbos et al., 2016; Athanassiou and Arthur, 2018).

281

Other natural substances or plant-derived insecticides, such as azadiractin, are also effective at

282

elevated concentrations that usually exceed 1000 ppm (Athanassiou et al., 2005). In fact, other

283

botanical extracts have been proved effective as contact insecticides against stored product

284

insects at even higher doses. Indicatively, Tofel et al. (2017), who examined various plant

285

tissues of A. indica and Plectranthus glandulosus Hook. (Lamiales: Lamiaceae), found that

286

satisfactory control of the cowpea weevil, Callosobruchus maculatus (F.) (Coleptera:

287

Bruchidae) and S. zeamais was possible at doses that exceeded 10000 ppm. At the same time,

288

there are some insecticides that have been proved particularly slow-acting against stored

289

product insect species due to their mode of action. For instance, the insect growth regulator

290

(IGR) S-methoprene affects only progeny production, despite the fact that in some cases there

291

is some parental mortality (Daglish et al., 2013). Similarly, the pyrrole chlorfenapyr causes a

12

292

delayed mortality and no knockdown after exposure, since its mode of action is sufficiently

293

different than that of neurotoxic insecticides (Dagg et al., 2019). Worth noting here is that the

294

epicuticular material of D. viscosa, as many other botanicals, may be easily manipulated

295

concerning the initial concentration of the bioactive substances. The present study used a

296

three-fold concentrated material in respect to the naturally occurring one on leaves but this

297

would be augmented by using higher amount of plant biomass in the same water volume,

298

resulting in more condensed and possibly more effective exudate.

299

There is extensive literature concerning flavonoid-phytophagous insect interactions

300

broadening our understanding on the influential role of flavonoids in host selection, survival,

301

growth, feeding behavior and reproduction of insects (Simmonds, 2003). Certain leaf

302

flavonoids directly interact with hormone system, or function as feeding deterrents, which is

303

also the case of sesquiterpenes (Gonzales-Coloma et al., 2010). Flavonoids on the leaf surface

304

act as the first barrier that the herbivore encounters after landing on the leaf and have been

305

reported to reduce the fitness of phytophagous insects, such as Euphydryas chalcedona

306

(Doubleday) (Lepidoptera: Nymphalidae) (Simmonds, 2003). In this context, the flavonoids

307

and sesquiterpenes present in D. viscosa epicuticular material may act as antifeedant agents

308

leading to starvation or exhibit moderate but direct toxic effects on the stored product insects

309

of the present study, thus further experiments needed in order to clarify their mode of action.

310

In summary, our results indicate that the lyophilized epicuticular material of D. viscosa has

311

some insecticidal value for the control of the species tested here, but this effect varies

312

according to the target species. In contrast with the majority of the studies that have evaluated

313

plant materials chiefly as formulated essential oils, the results of the present work show that

314

the D. viscosa material used provided some insecticidal effect when admixed with the grain,

315

suggesting that its mode of action can be comparable with that of other contact insecticides.

316

Further research is needed to shed light to these parameters, including the mode of action of

13

317

D. viscosa epicuticular material, and to evaluate the basis of its use in realistic scenarios in

318

stored-product protection.

319 320

References

321

Aitken A. 1975. Insect Travelers, I: Coleoptera, Techn. Bull. 31, H.M.S.O. London.

322

Allal-Benfekih, L., Bellatreche, M., Bounaceur, F., Tail, G., Mostefaoui, H., 2011. First

323

approach of using aqueous extracts of Inula viscosa, Salvia officinalis and Urtica urens

324

for the control of Tuta absoluta (Lepidoptera, Gelechiidae) an invasive pest of tomato in

325

Algeria. Scale insects: main or secondary pest. 9th International Conference on Pests in

326

Agriculture. SupAgro, Montpellier, Fr. 25–27 Octobre 2011. pp. 681–689

327 328

Arthur, F. H., 1996. Grain protectants: Current status and prospects for the future. J. Stored Prod. Res. 32, 293-302.

329

Arthur, F. H., Throne, J. E., 2003. Efficacy of diatomaceous earth to control internal

330

infestations of rice weevil and maize weevil (Coleoptera: Curculionidae). J. Econ.

331

Entomol. 96, 510-518.

332 333

Athanassiou, C. G., Arthur, F. H., 2018. Recent Advances in Stored Product Protection, pp. 273.

334

Athanassiou, C. G., Kontodimas, D. C., Kavallieratos, N. G., Veroniki, M. A., 2005.

335

Insecticidal effect of NeemAzal against three stored-product beetle species on rye and

336

oats. J. Econ. Entomol. 98, 1733-1738.

337 338

Athanassiou, C. G., Usha Rani, P., Kavallieratos, N. G., 2014. The use of plant extracts for stored product protection. In: Advances in Plant Biopesticides, pp. 131-147.

14

339

Batta, Y. A., 2005. Control of the lesser grain borer (Rhyzopertha dominica (F.), Coleoptera:

340

Bostrichidae) by treatments with residual formulations of Metarhizium anisopliae

341

(Metschnikoff) Sorokin (Deuteromycotina: Hyphomycetes). J. Stored Prod. Res. 41, 221-

342

229.

343

Boussaada, O., Ben Halima Kamel, M., Ammar, S., Haouas, D., Mighri, Z., Helal, A. N., 2008.

344

Insecticidal activity of some asteraceae plant extracts against Tribolium confusum. Bull.

345

Insectology 61, 283-289.

346

Dagg, K., Irish, S., Wiegand, R. E., Shililu, J., Yewhalaw, D., Messenger, L. A., 2019.

347

Evaluation of toxicity of clothianidin (neonicotinoid) and chlorfenapyr (pyrrole)

348

insecticides and cross-resistance to other public health insecticides in Anopheles

349

arabiensis from Ethiopia. Malar. J. 18, 1-11.

350 351

Daglish, G. J., Holloway, J. C., Nayak, M. K., 2013. Implications of methoprene resistance for managing Rhyzopertha dominica (F.) in stored grain. J Stored Prod. Res. 54, 8-12.

352

Daglish, G. J., Nayak, M. K., Arthur, F. H., Athanassiou, C. G., 2018. Insect pest management

353

in stored grain. In: Athanassiou C. G., Arthur F H. (eds.), Recent Advances in Stored

354

Product Protection, Springer, pp. 45-63.

355

Daniewski, W.M., Kroszczynski, W., Bloszyk, E., Drozdz, B., Nawrot, J., Rychlewska, U.,

356

Budesinsky, M., Holub, M., 1986. On terpenes. 294. Sesquiterpenoids from Dittrichia

357

viscosa (L.) Greuter. Their structure and deterrent activity. Collect. Czech. Chem.

358

Commun. 51, 1710-1721.

359 360

Dubey, N. K., Shukla, R., Kumar, A., Singh, P., Prakash, B., 2010. Prospects of botanical pesticides in sustainable agriculture. Curr. Sci. 98, 479-480.

15

361

Grauso, L., Cesarano, G., Zotti, M., Ranesi, M., Sun, W., Bonanomi, G., Lanzotti, V., 2019.

362

Exploring Dittrichia viscosa (L.) greuter phytochemical diversity to explain its

363

antimicrobial, nematicidal and insecticidal activity. Phytochem. Rev. pp. 1-31.

364

Gonzalez-Coloma, A., Reina, M., Diaz, C.E., Fraga, B.M., 2010. Natural Product-Based

365

Biopesticides for Insect Control. Comprehensive Natural Products II 3, pp. 237-268.

366

Haines, C.P., 1991. Insects and Arachnids of Tropical Stored Products: Their Biology and

367

Identification (A Training Manual), Natural Resources Institute, Ed. 2, pp. 246.

368

Hernandez-Lambraño, R., Pajaro-Castro, N., Caballero-Gallardo, K., Stashenko, E., Olivero-

369

Verbel, J., 2015. Essential oils from plants of the genus Cymbopogon as natural

370

insecticides to control stored product pests. J. Stored Prod. Res. 62, 81-83.

371

Huang, Y., Lam, S.L., Ho, S.H., 2000. Bioactivities of essential oil from Elletaria cardamomum

372

(L.) Maton. to Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst). J.

373

Stored Prod. Res. 36, 107–117.

374

Jagadeesan, R., Collins, P. J., Daglish, G. J., Ebert, P. R., Schlipalius, D. I., 2012. Phosphine

375

resistance in the rust red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae):

376

Inheritance, gene interactions and fitness costs. PLoS ONE 7, e31582.

377

Jagadeesan, R., Fotheringham, A., Ebert, P. R., Schlipalius, D. I., 2013. Rapid genome wide

378

mapping of phosphine resistance loci by a simple regional averaging analysis in the red

379

flour beetle, Tribolium castaneum. BMC Genomics 14, 650.

380

Kaur, R., Daniels, E. V., Nayak, M. K., Ebert, P. R., Schlipalius, D. I., 2013. Determining

381

changes in the distribution and abundance of a Rhyzopertha dominica phosphine

16

382

resistance allele in farm grain storages using a DNA marker. Pest Manag. Sci. 69, 685-

383

688.

384 385

Korunic, Z., 1998. Diatomaceous earths, a group of natural insecticides. J. Stored Prod. Res. 34, 87-97.

386

Lee, B., Lee, S., Annis, P. C., Pratt, S. J., Park, B., Tumaalii, F., 2002. Fumigant toxicity of

387

essential oils and monoterpenes against the red flour beetle, Tribolium castaneum Herbst.

388

J. Asia Pac. Entomol. 5, 237-240.

389

Levizou, E., Karageorgou, P., Psaras, G.K., Manetas, Y., 2002. Inhibitory effects of water

390

soluble leaf leachates from Dittrichia viscosa on lettuce root growth, statocyte

391

development and graviperception. Flora 197, 152–157.

392

Mamoci, E., Cavoski, I., Andres, M.F., 2012. Chemical characterization of the aphid

393

antifeedant extracts from Dittrichia viscosa and Ferula communis. Biochem. Syst. Ecol.

394

43, 101-107.

395

Mayhew, T. J., Phillips, T. W., 1994. Pheromone biology of the lesser grain borer, Rhyzopertha

396

dominica (Coleoptera: Bostrichidae). In Proceedings of the 6th International Working

397

Conference on Stored Product Protection (Edited by Highley E., Wright E. J., Banks H. J.

398

and Champ B. R.) pp. 541-544. Canberra, Australia.

399

Nayak, M. K., Daglish, G. J., 2018. Importance of stored product insects. In: Athanassiou C. G.,

400

Arthur F H. (eds.), Recent Advances in Stored Product Protection, Springer, pp. 1-17.

401

Oka, Y., Ben-Daniel, B.-H., Cohen, Y., 2001. Nematicidal activity of powder and extracts of

402

Inula viscosa. Nematology 3, 735-742.

17

403 404

405 406

Parolin, P., Ion Scotta, M., Bresch, C., 2014. Biology of Dittrichia viscosa, a Mediterranean ruderal plant. J. Exp. Bot. 83, 251-262. Pascual-Villalobos, M. J., Robledo, A., 1998. Screening for anti-insect activity in mediterranean plants. Ind. Crop Prod. 8, 183-194.

407

Pavela, R., 2004. Insecticidal activity of certain medicinal plants. Fitoterapia 75, 745-749.

408

Prakash A., Rao J., 1997. Botanical pesticides in agriculture, CRC Press, pp. 1-364.

409

Qasem, J. R., Abu‐Blan, H. A., 1995. Antifungal activity of aqueous extracts from some

410

411

common weed species. Ann. Appl. Biol. 127, 215-219. Rita Devi, S., Thomas, A., Rebijith, K. B., Ramamurthy, V. V., 2017. Biology, morphology and

412

molecular characterization of Sitophilus oryzae and S. zeamais

413

Curculionidae). J. Stored Prod. Res. 73, 135-141.

(Coleoptera:

414

Rojht, H., Horvat, A., Athanassiou, C. G., Vayias, B. J., Tomanović, Ž., Trdan, S., 2010. Impact

415

of geochemical composition of diatomaceous earth on its insecticidal activity against

416

adults of Sitophilus oryzae (L.) (Coleoptera: Curculionidae). J. Pest Sci. 83, 429-436.

417

Rotundo, G., Paventi, G., Barberio, A., De Cristofaro, A., Notardonato, I., Russo, M. V.,

418

Germinara, G. S., 2019. Biological activity of Dittrichia viscosa (L.) greuter extracts

419

against adult Sitophilus granarius (L.) (Coleoptera: Curculionidae) and identification of

420

active compounds. Sci. Rep. 9, 6429.

421

Rumbos, C. I., Sakka, M., Berillis, P., Athanassiou, C. G., 2016. Insecticidal potential of zeolite

422

formulations against three major stored-grain insects: particle size effect, adherence ratio

423

and effect of test weight on grains. J. Stored Prod. Res. 68, 93-101.

18

424

Schlipalius, D. I., Cheng, Q., Reilly, P. E. B., Collins, P. J., Ebert, P. R., 2002. Genetic linkage

425

analysis of the lesser grain borer Rhyzopertha dominica identifies two loci that confer

426

high-level resistance to the fumigant phosphine. Genetics 161, 773-782.

427 428

429 430

Simmonds M.S.J., 2003. Flavonoid–insect interactions: recent advances in our knowledge. Phytochem. 64, 21-30. Sokal, R.R., Rohlf, F.J., 1995. Biometry: The Principles and Practice of Statistics in Biological Research. Stony Brook University.

431

Stavrianakou, S., Liakoura, V., Levizou, E., Karageorgou, P., Delis, C., Liakopoulos, G.,

432

Karabourniotis, G., Manetas, Y., 2004. Allelopathic effects of water-soluble leaf

433

epicuticular material of Dittrichia viscosa on seed germination of crops and weeds.

434

Allelopathy J. 14, 35-42.

435

Stejskal, V., Hubert, J., Li, Z., 2018. Human health problems and accidents associated with

436

occurrence and control of storage arthropods and rodents. Insect pest management in

437

stored grain. In: Athanassiou C. G., Arthur F H. (eds.), Recent Advances in Stored

438

Product Protection, Springer, pp. 19-43.

439

Stephanou, M., Manetas, Y., 1997. Seasonal variations in UV-B absorbing capacity and

440

allelopathic potential of Dittrichia viscosa leaf rinsates. Can. J. Bot. 75, 1371-1374.

441

Tapondjou, A. L., Adler, C., Fontem, D. A., Bouda, H., Reichmuth, C., 2005. Bioactivities of

442

cymol and essential oils of Cupressus sempervirens and Eucalyptus saligna against

443

Sitophilus zeamais Motschulsky and Tribolium confusum du val. J. Stored Prod. Res. 41,

444

91-102.

445

Tofel, K.H., Kosma, P., Stahler, M., Adler, C., Nukenine, E.N., 2017. Insecticidal products

446

from Azadirachta indica and Plectranthus glandulosus growing in Cameroon for the 19

447

protection of stored cowpea and maize against their major insect pests. Ind. Crop Prod.

448

110, 58–64.

449

Topakci, N., Ikten, C., Gosmen, H., 2005. A research on some effects of Inula viscosa (L.) Ait.

450

(Asteraceae) leaf extract on carmine spider mite, Tetranychus cinnabarinus (Boisd.)

451

(Acari:Tetranychidae). Akdeniz Universitesi Ziraat Fakultesi Dergisi, 18, 411-415.

452

Torres, P., Avila, J. G., Romo De Vivar, A., Garcia A. M., Marin J. C., Aranda, E., Cespedes,

453

C. L., 2003. Antioxidant and insect growth regulatory activities of stilbenes and extracts

454

from Yucca periculosa. Phytochem. 64, 463- 473.

455 456

Tunç, I., Berger, B. M., Erler, F., Dagli, F., 2000. Ovicidal activity of essential oils from five plants against two stored-product insects. J. Stored Prod. Res. 36, 161-168.

457

Weaver, D.K., Subramanyam, B., 2000. Botanicals. In: Subramanyam B, Hagstrum DW (eds)

458

Alternatives to pesticides in stored-product IPM. Kluwer Academic Publishers,

459

Dordrecht, pp. 303–320.

460

Yegen, O., Berger, B., Heitefuss, R., 1992. Untersuchungen zur fungitoxischen wirkung der

461

extrakte sechs ausgewahlter pflanzen aus der Turkei auf phytopathogene pilze. Anz.

462

Pflanzenkr. Pflanzenschutz. 99, 349-359.

463 464 465 466 467 468

20

469 Table 1. Repeated Measures MANOVA parameters for mortality levels of the four species tested (total df=3

R. dominica S. oryzae df F F F P Between variables 3 0.37 0.77 4.33 0.01 Y- intercept 1 11.81 <0.01 56.67 <0.01 Dose 3 0.37 0.77 4.33 0.01 Within variables 12 0.97* 0.47 2.40* 0.01 Exposure 4 5.08 <0.01 23.74 <0.01 Exposure X Dose 12 0.97* 0.47 2.40* 0.01 470 * Wilks’ Lamda approximate F value 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486

21

Species T. confusum F P 17.08 <0.01 85.15 <0.01 17.08 <0.01 8.00* <0.01 43.90 <0.01 8.00* <0.01

O. surinamensis F P 69.56 <0.01 264.68 <0.01 69.56 <0.01 8.88* <0.01 61.19 <0.01 8.88* <0.01

487 Table 2. Mean mortality (% ± SE) of R. dominica adults after 1, 3, 7, 14 and 21 days of exposure on 488 wheat treated with different concentrations of the D. viscosa formulation (in all cases df=3.35).

Exposure interval Treatment

1d

3d

7d

14 d

21 d

Control

0.6 ± 0.6

0.6 ± 0.6

1.6 ± 1.1

2.2 ± 1.2

3.9 ± 1.1

1000 ppm

0.6 ± 0.6

0.6 ± 0.6

1.1 ± 1.1

9.4 ± 5.8

11.6 ± 7.4

3000 ppm

2.2 ± 1.2

3.3 ± 2.2

5.0 ± 2.7

5.0 ± 2.7

5.6 ± 2.7

5000 ppm

2.2 ± 2.2

3.3 ± 2.2

3.8 ± 2.3

5.0 ± 2.7

7.2 ± 3.2

F

0.52

0.99

0.86

0.69

0.60

P

0.66

0.40

0.47

0.56

0.61

489 Within each column, means followed by the same letter are not significantly different (Tukey-Kramer 490 HSD test at 0.05; where no letters exist, no significant differences were noted). 491 492 493 494 495 496 497 498 499 500 501 502 503 504 22

505 Table 3. Mean mortality (% ± SE) of S. oryzae adults after 1, 3, 7, 14 and 21 days of exposure on 506 wheat treated with different concentrations of the D. viscosa formulation (in all cases df=3.35).

Exposure interval Treatment

1d

3d

7d

14 d

21 d

Control

0.6 ± 0.6

1.1 ± 0.7 b

3.3 ± 1.4 b

6.6 ± 2.9 b

11.6 ± 3.3 b

1000 ppm

1.1 ± 0.7

2.7 ± 1.6 ab

10.0 ± 3.9 ab

12.2 ± 3.9 ab

13.9 ± 3.7 ab

3000 ppm

0.0 ± 0.0

8.3 ± 1.6 a

25.0 ± 5.3 a

26.6 ± 5.2 a

32.2 ± 5.3 a

5000 ppm

0.6 ± 0.6

5.6 ± 2.4 ab

23.3 ± 6.8 a

24.4 ± 7.1 ab

26.6 ± 6.8 ab

F

0.71

3.35

4.69

3.63

3.89

P

0.55

0.03

<0.01

0.02

0.01

507 Within each column, means followed by the same letter are not significantly different (Tukey-Kramer 508 HSD test at 0.05; where no letters exist, no significant differences were noted). 509 510 511 512 513 514 515 516 517 518 519 520 521

23

522 Table 4. Mean mortality (% ± SE) of T. confusum adults after 1, 3, 7, 14 and 21 days of exposure on 523 wheat treated with different concentrations of the D. viscosa formulation (in all cases df=3.35).

Exposure interval Treatment

1d

3d

7d

14 d

21 d

Control

0.0 ± 0.0

0.6 ± 0.6

0.6 ± 0.6 b

2.7 ± 1.2 c

2.7 ± 1.2 c

1000 ppm

0.0 ± 0.0

0.6 ± 0.6

2.2 ± 1.2 ab

3.3 ± 1.1 c

6.1 ± 1.3 c

3000 ppm

0.0 ± 0.0

1.1 ± 0.7

6.6 ± 2.2 a

13.3 ± 4.2 b

17.2 ± 4.0 b

5000 ppm

1.1 ± 1.1

1.1 ± 1.1

3.9 ± 1.8 ab

26.1 ± 2.4 a

35.6 ± 3.2 a

F

0.99

0.17

2.72

17.75

28.30

P

0.40

0.91

<0.01

<0.01

<0.01

524 Within each column, means followed by the same letter are not significantly different (Tukey-Kramer 525 HSD test at 0.05; where no letters exist, no significant differences were noted). 526 527 528 529 530 531 532 533 534 535 536 537 538

24

539 Table 5. Mean mortality (% ± SE) of O. surinamensis adults after 1, 3, 7, 14 and 21 days of exposure 540 on wheat treated with different concentrations of the D. viscosa formulation (in all cases df=3.35).

Exposure interval Treatment

1d

3d

7d

14 d

21 d

Control

0.0 ± 0.0

0.0 ± 0.0 b

0.6 ± 0.6 c

3.3 ± 1.4 c

6.6 ± 2.2 c

1000 ppm

0.6 ± 0.6

3.9 ± 1.6 ab

5.6 ± 1.5 c

7.7 ± 1.6 c

11.6 ± 2.9 c

3000 ppm

1.1 ± 0.7

11.1 ± 3.3 a

35.0 ± 7.8 b

43.9 ± 6.9 b

46.1 ± 7.4 b

5000 ppm

0.6 ± 0.6

13.9 ± 3.9 a

58.3 ± 6.5 a

84.4 ± 4.1 a

86.6 ± 4.7 a

F

0.71

5.70

27.10

81.66

60.32

P

0.55

<0.01

<0.01

<0.01

<0.01

541 Within each column, means followed by the same letter are not significantly different (Tukey-Kramer 542 HSD test at 0.05; where no letters exist, no significant differences were noted). 543 544 545 546 547 548 549 550 551 552 553 554 555 556 25

557 Table 6. ANOVA parameters for progeny production counts for each species (total df=32).

Species O. surinamensis

T. confusum

S. oryzae

R. dominica

df

F

P

F

P

F

P

F

P

Model

3

8.78

<0.01

3.96

0.01

1.36

0.27

8.33

<0.01

Y-Intercept

1

33.95

<0.01

18.46

<0.01

102.45

<0.01

57.28

<0.01

Dose

3

8.78

<0.01

3.96

<0.01

1.36

0.27

8.33

<0.01

558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574

26

575 Table 7. Mean number of progeny production (adults per vial ± SE) for each species on treated and 576 untreated wheat, 65 d after the removal of the parental adults (in all cases df=3.35).

Species Treatment

O. surinamensis

T. confusum

S. oryzae

R. dominica

Control

40.0 ± 10.6 a

4.6 ± 1.4 a

73.9 ± 8.7

107.4 ± 22.9 a

1000 ppm

37.7 ± 9.0 a

1.5 ± 0.6 ab

77.0 ± 15.2

69.0 ± 15.9 ab

3000 ppm

4.9 ± 2.6 b

0.9 ± 0.6 b

84.7 ± 20.2

21.1 ± 4.2 b

5000 ppm

0.2 ± 0.2 b

0.7 ± 0.4 b

47.2 ± 8.0

21.3 ± 6.4 b

F

0.78

3.96

1.36

8.33

P

<0.01

0.01

0.27

<0.01

577

Within each column, means followed by the same letter are not significantly different (Tukey-

578

Kramer HSD test at 0.05; where no letters exist, no significant differences were noted).

579

27

1

Crop Protection

Prof. Christos G. Athanassiou

2

Laboratory of Entomology and

3

Agricultural Zoology

4

University of Thessaly

5

Phytokou str., 38446, Nea Ionia

6

Magnessia, Greece

7

Phone: +30-2421093195

8

FAX: +30-2421093178

9

10

Insecticidal Effect of Dittrichia viscosa lyophilized epicuticular material against

11

four major stored-product beetle species on wheat

12

E. Lampiri a,b, P. Agrafioti a, E. Levizou b, C.G. Athanassiou a*

13

14

a

15

Crop Production and Rural Environment, University of Thessaly, Phytokou str., Nea

16

Ionia, Magnesia, 38446, Greece

17

b

18

Rural Environment, University of Thessaly, Phytokou str., Nea Ionia, Magnesia,

19

38446, Greece

20

(* corresponding author, e-mail address: [email protected])

Laboratory of Entomology and Agricultural Zoology, Department of Agriculture,

Laboratory of Weed Science, Department of Agriculture, Crop Production and

21

[1]

22

23

Research Highlights •

material of Dittrichia viscosa for the control of stored-grain insect species.

24 25

This study is the first that examined the insecticidal effect of epicuticular

•

The lyophilized epicuticular material of D. viscosa has some insecticidal value

26

for the control of the species tested, but this effect varies according to the target

27

species.

28

•

Oryzaephilus surinamensis was found to be the most susceptible, followed by Tribolium confusum and Sitophilus oryzae.

29 30

•

Rhyzopertha dominica was found to be the most resistant species.

31

•

Progeny production was particularly reduced for all species relative to the

32

controls.

[2]