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Arthropod-Plant InteractionsAn international journal devoted tostudies on interactions of insects, mites,and other arthropods with plants ISSN 1872-8855 Arthropod-Plant InteractionsDOI 10.1007/s11829-012-9236-x

Biological activities of lignans andneolignans on the aphid Myzus persicae(Sulzer)

Julien Saguez, Jacques Attoumbré,Philippe Giordanengo & Sylvie Baltora-Rosset

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ORIGINAL PAPER

Biological activities of lignans and neolignans on the aphidMyzus persicae (Sulzer)

Julien Saguez • Jacques Attoumbre •

Philippe Giordanengo • Sylvie Baltora-Rosset

Received: 14 November 2011 / Accepted: 31 October 2012

� Springer Science+Business Media Dordrecht 2012

Abstract Lignans and neolignans have been reported to

exert different biological activities, including insecticidal

ones. Three lignans, secoisolariciresinol (SECO), secoisol-

ariciresinol diglucoside (SDG), and anhydrosecoisolaricir-

esinol (AHS), and one neolignan, dehydrodiconiferyl

alcohol-4-b-D-glucoside (DCG), were isolated from flax.

Their insecticidal properties were evaluated on the aphid

Myzus persicae reared on artificial diet. Life history

parameters, i.e., nymphal survival, prereproductive period,

and daily fecundity, were assessed and used to calculate the

intrinsic rate of natural increase and the doubling time of

aphid populations. Compared to the control, SDG and DCG

significantly increased aphid mortality by at least 25 %,

while SECO and AHS did not affect their survival. SDG did

not affect life history parameters, except at the highest

concentration of 100 lg/mL, which increased the popula-

tion’s doubling time by more than 5 days. DCG altered all

the life history parameters at all concentrations assayed.

SECO induced significant deleterious effects on the aphids,

except at the highest concentration of 100 lg/mL. AHS only

altered prereproductive period, which increased by at least

2 days at 50 and 100 lg/mL. Lignans and neolignans are

potential new bioinsecticides against aphids in the context of

alternative management programs.

Keywords Aphid � Flax � Lignans � Neolignans �Artificial diet � Demographic parameters

Introduction

In plants, the oxidative dimerization of two phenylpropa-

noid units leads to a great variety of phytochemicals des-

ignated as lignans and neolignans. When the oxidative

process implies the 8–80 bonds, the metabolites are named

lignans, while the term neolignans is used for those issued

from all other types of linkage. Lignans and neolignans

constitute a large group of phenolic compounds widely

distributed in the plant kingdom and they show a large

chemical structural diversity. Lignans are found in almost

all the parts of plants, including roots, leaves, flowers,

fruits, and seeds (Willfor et al. 2006). Although their bio-

logical significance is complex, their wide occurrence in

nature has induced numerous investigations in food science

and for their pharmacological properties (e.g., antitumor

activity) in mammalian systems (MacRae and Towers

1984; Moujir et al. 2007; Pool-Zobel et al. 2000; Saleem

et al. 2005). Moreover, these natural phytochemicals can be

used as templates for the development of potential new

botanical agents with biological activities for controlling

Handling Editors: Yvan Rahbe and Heikki Hokkanen.

J. Saguez � J. Attoumbre � P. Giordanengo � S. Baltora-Rosset

Universite de Picardie Jules Verne, 33 Rue St Leu,

80039 Amiens cedex 1, France

e-mail: Jacques.attoumbre@u-picardie.fr

P. Giordanengo

e-mail: pgiordanengo@sophia.inra.fr

S. Baltora-Rosset

e-mail: Sylvie.baltora-rosset@u-picardie.fr

J. Saguez (&)

Agriculture and Agri-Food Canada, Horticultural Research and

Development Centre, 430 Gouin Boulevard,

Saint-Jean-sur-Richelieu, QC J3B 3E6, Canada

e-mail: julien.saguez@u-picardie.fr

P. Giordanengo

Institut Sophia Agrobiotech, CNRS 7254, INRA 1355,

Universite de Nice Sophia Antipolis, 400 route des Chappes,

BP167, 06903 Sophia Antipolis, France

123

Arthropod-Plant Interactions

DOI 10.1007/s11829-012-9236-x

Author's personal copy

various pests. Lignans are known to be important chemical

plant defences that are involved in allelopathic interactions

(Dixon 2004; MacRae and Towers 1984; Saraiva et al.

2007) to fight microorganisms such as viruses, bacteria,

fungi (Pauletti et al. 2000; Vargas-Arispuro et al. 2005;

Willfor et al. 2006), and phytophagous animals (e.g.,

nematodes, molluscs, vertebrates).

Insects respond to a large range of phytochemicals,

which may alter their physiological performance by dis-

rupting mechanical, physical, biological, and biochemical

functions related to behavior, development, and reproduc-

tion (Harmatha and Dinan 2003). These chemicals of plant

origin receive widespread interest since concerns for

human and environmental health have led to banish or

severely restrict the use of synthetic insecticides in many

countries worldwide (Scott et al. 2008). However, the

effects of lignans and neolignans on insects and their role

in plant–insect interactions are poorly documented. Only

few studies have reported general intoxication or feeding

deterrence or developmental disruption (Garcia and Aza-

mbuja 2004; Harmatha and Dinan 2003). Insecticidal

activity has been reported on coleopteran (Harmatha and

Nawrot 2002) and dipteran pests (Harmatha and Dinan

2003) and is mainly associated with antifeedant effects.

Lignans have also been reported to induce molting distur-

bances (Garcia and Azambuja 2004) and mortality (Cabral

et al. 2000; Messiano et al. 2008; Russell et al. 1976).

Aphids are key insect pests of northern European agri-

culture (Hulle et al. 2010), causing serious losses to culti-

vated crops by phloem sap withdrawal and indirect losses

through honeydew production favouring the development

of sooty molds and transmission of viral diseases. Myzus

persicae, one of the most common and polyphagous aphids

worldwide, is a vector of numerous viruses that reduce

yields and depreciate the quality of crops (Blackman and

Eastop 1984). Because no direct antiviral treatment is

available, the only strategy to fight virus diseases, except

the genetic improvement of plant resistance, is to manage

aphid populations. Aphid infestations can be managed by

insecticides, often resulting in the development of resistant

populations (Devonshire et al. 1998). Alternatively, several

studies are in progress to develop strategies to manage

pathogen and pest populations, including methods which

take advantage of natural chemicals produced by plants

(Carlini and Grossi-de-Sa 2002; Regnault-Roger et al.

2005). With the exception of Macrosiphum euphorbiae that

induce yield losses (Wise et al. 1995), only few aphid

species (Acyrthosiphon ilka, Acyrthosiphon mordvilkoi,

Aphis craccivora, Aphis fabae and Linaphis lini) have been

reported to develop on flax (Linum usitatissimum) (Fergu-

son et al. 1997; Lamb and Grenkow 2008; Wise et al. 1995;

Blackman and Eastop 1984). Although M. persicae is a

highly polyphagous species, its presence on flax has not

been mentioned yet. This suggests potential protective/

repulsive or deleterious effects of flax phytochemicals on

aphids.

Flaxseed presents one of the highest known lignan

contents, estimated to be 60–700 times greater than in other

plants. The main lignan is secoisolariciresinol (SECO)

which accumulates in its diglucoside form (secoisolaric-

iresinol diglucoside, SDG) (Ford et al. 2001). Furthermore,

the two compounds, anhydrosecoisolariciresinol (AHS)

and dehydrodiconiferyl alcohol-4-b-D-glucoside (DCG),

could exert biological activities or be involved in plant

resistance (Hano et al. 2006; Lehraiki et al. 2010).

Immunolocalization experiments combined with HPLC

analysis indicated that 90 % of the lignans are localized in

the outer integument of seeds (11.43 mg/g dry weight) and

that they were also present in flax stems in small quantity

(0.3 mg/g dry weight) (Attoumbre et al. 2010). The pres-

ence of lignans in flax stems suggests that aphids can be

exposed to these compounds. It can also explain the in vivo

potential impacts of lignans in flax, such as the low

infestation by aphids or their absence on this crop, due to

deterrent, antifeedant, or toxic effects.

We evaluated these four compounds (SECO, SDG,

AHS, and DCG) on aphid mortality and demographic

parameters. The fine chemical variations concerning

structural features such as hydroxyl and carbohydrate

moieties could help to point out the role of these functional

groups on biological activities in aphids. According to our

knowledge, this study constitutes the first report on the

biological effects of lignans on aphids.

Materials and methods

Lignan and neolignan extractions

Three lignans and one neolignan (Fig. 1) were isolated and

purified from flax as described by Lehraiki et al. (2010). L.

usitatissimum seeds (cv. Barbara) were used to isolate

secoisolariciresinol diglucoside (SDG) which is the storage

form of secoisolariciresinol (SECO) in flax seeds. The

anhydrous form of secoisolariciresinol aglycone, the

anhydrosecoisolariciresinol (AHS), was obtained in high

acidic conditions in the following conditions: 340 g of

defatted flaxseed were refluxed for 75 min in 2.4 L of

water with 235 mL of concentrated hydrochloric acid.

After cooling the solution was extracted three times with

EtOAc/hexane (90/10, v/v) and the combined organic

phases were evaporated to dryness. The residue (about

24 g) was purified by silica-gel column chromatography

and fractionated by elution with various proportions of

EtOAc and hexane. Anhydrosecoisolariciresinol was eluted

in EtOAc/hexane (20/80, v/v). Secoisolariciresinol was

J. Saguez et al.

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eluted in EtOAc/hexane (70/30, v/v). The fractions were

analyzed by thin layer chromatography and combined

when having the same Rf values. These fractions were

further purified by semi-preparative high pressure liquid

chromatography (HPLC). A neolignan, the dehydrodico-

niferyl alcohol-4-b-D-glucoside (DCG), was extracted from

roots cell cultures established from L. usitatissimum using a

protocol adapted from Attoumbre et al. (2006) and briefly

described hereafter. A freeze-dried and ground sample of

cells (5 g) cultured on Murashige and Skoog-derived

medium was extracted with methanol/water (70/30, v/v) for

3 h at 60 �C. After filtration, the extract was concentrated.

DCG was separated and purified by semi-preparative

HPLC.

Insect bioassays

The M. persicae (Sulzer) colony was maintained in a

growth chamber, on potato plants (Solanum tuberosum cv

Desiree), under parthenogenesis-inducing conditions, i.e.,

20 ± 1 �C and L16:D8 photoperiod (Saguez et al. 2006).

Females of M. persicae were placed on a standard artificial

diet to obtain synchronized offsprings. Cages were built

using PVC tube (3 cm diameter, 1 cm height) and double-

layer Parafilm� pouches containing 100 lL of a standard

diet adapted for M. persicae (Down et al. 1996). Briefly,

the standard diet prepared under aseptic conditions was

composed of a solution of ten vitamins, all the essential and

nonessential amino acids, saccharose (20 %), and traces of

metals and other elements. This standard diet was used as a

control and as a carrier for lignan dilutions. Lignans and

neolignans were previously solubilized at 1 mg/mL in 1 %

methanol before their dilution in the standard diet to obtain

the final concentrations of 1, 10, 50, and 100 lg/mL. Ten

young (\24 h) nymphs were transferred to each cage, put

on pouches, and maintained under the same rearing con-

ditions as described above. At least five replicates were

carried out for each diet, and the pouches were changed

every second day.

Nymphal survival and adult emergence were daily

recorded. Aphids that reached adulthood the same day were

pooled and isolated on new pouches to evaluate their pre-

reproductive period (PRP) (i.e., elapsed time from first

nymphal instar until onset of reproduction) and their daily

Fig. 1 Chemical structures of secoisolariciresinol (SECO), secoisol-

ariciresinol diglucoside (SDG), anhydrosecoisolariciresinol (AHS),

and dehydrodiconiferyl alcohol-4-b-D-glucoside (DCG) isolated from

L. usitatissimum. For DCG, chirality is not indicated because

stereochemistry was not fully determined

Biological activities of lignans and neolignans

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fecundity (DF) until the death of the aphids. The intrinsic rate

of natural increase (rm) and the doubling time (DT) of M.

persicae populations were calculated according to Le Roux

et al. (2004) with DEMP software (Giordanengo 2012).

Statistical analysis

The effects of lignans on nymphal survival were analyzed

with Pearson’s v2 test. One-way analysis of variance

(ANOVA) was carried out with Statistica 5.5 software

(StatSoft�, Tulsa, Oklahoma, USA) to test the effects of the

diets on aphid demographic parameters. Significantly altered

demographic parameters were further analyzed with Fisher’s

positive least significant difference test (PLSD) at P \ 0.05.

Results

Nymphal survival of M. persicae (Table 1)

Four lignans and neolignans extracted from flax, i.e., the

secoisolariciresinol diglucoside (SDG), the secoisolaric-

iresinol (SECO), the anhydrosecoisolariciresinol (AHS),

and the dehydrodiconiferyl alcohol-4-b-D-glucoside (DCG)

(Fig. 1) were tested on M. persicae, to evaluate their

potential aphicidal effects. None of the tested four con-

centrations, i.e., 1, 10, 50, or 100 lg/mL of SECO and

AHS affected nymphal survival.

DCG did not induce any effect at 1 and 50 lg/mL, while

nymphal survival was significantly reduced to 20 and 33 %

at 10 and 100 lg/mL, respectively.

Nymphal survival of M. persicae was also significantly,

respectively, reduced to 15, 22.5, and 32.5 % when SDG

was added to the diet at 10, 50, and 100 lg/mL when

compared with the control diet, while no difference was

observed at 1 lg/mL.

Demographic parameters (Table 2)

At all concentrations, DCG increased PRP by more than

1 day. A reduction of ca. 25 % of daily fecundity (DF) was

also observed at the concentrations lower than 100 lg/mL.

Consequently, the intrinsic rate of natural increase (rm)

values were significantly reduced and the doubling time of

populations (DT) was increased for all concentrations.

SDG did not induce any effect at 1 and 10 lg/mL when

compared with the control diet. DF significantly increased

at 50 lg/mL. However, this increase did not affect rm value

and DT, due to the high mortality recorded at this con-

centration. At 100 lg/mL, the rm value decreased and the

DT was increased by more than 5 days.

AHS increased PRP at the highest concentrations of 50

and 100 lg/mL and DF at 1 lg/mL, and reduced the rm

value at 50 lg/mL. The other parameters did not differ

from the control.

Although PRP was not modified whatever the SECO

concentrations, DF and rm values were reduced to 1, 10,

and 50 lg/mL, and DT lengthened by approximately one-

third. No effect was observed at 100 lg/mL.

Discussion

Biological effects of lignans on M. persicae

DCG, SDG, SECO, and AHS provided as food through

artificial diet to M. persicae had variable deleterious effects

that can reflect their direct or indirect effects on aphid

mortality, physiology, and growth parameters.

Nymphal survival was only altered by the glycosylated

DCG and SDG, which caused approximately 30 % nym-

phal mortality at 10 and 100 lg/mL for DCG and at 10, 50,

and 100 lg/mL for SDG. In contrast, no significant effect

on nymphal survival was observed for aphids fed on the

artificial diet supplemented with aglycone lignans (SECO

and AHS). The increase of mortality could be assigned to

three different mechanisms due to (1) antifeedant or

deterrent effects, (2) intoxication, and (3) indirect effects

on the endosymbionts Buchnera sp.

Antifeedant and deterrent activities of lignans were also

reported on coleopteran (Harmatha and Nawrot 2002),

dipteran (Harmatha and Dinan 2003), and lepidopteran

pests (Messiano et al. 2008; Nawrot et al. 1991). Further-

more, antifeedant and deterrent effects were also observed

on hemiptera (Cabral et al. 1999). In the field, aphids can

be exposed to lignans present in stems (Attoumbre et al.

2010) and fibers (Krajcova et al. 2009). Aphids are able to

detect antifeedant or deterrent compounds (Messchendorp

et al. 1998) which can induce their flight to find a most

suitable host to feed and could at least partly explain why

aphid colonies do not develop on flax (Ferguson et al.

1997). In contrast, in our study, aphids are forced to feed on

the artificial diet to survive. The reduction of nymphal

survival could be explained by the antifeedant effects of

lignans and neolignans that cause cessation of food intake,

starvation, and eventually death.

Lignans and neolignans can directly affect aphids by

intoxication after ingestion. Russel et al. (1976) reported

toxic effects of lignans on Musca domestica (Diptera) and

Laspeyresia pomonella (Lepidoptera). Another study

reported that lignans induced high mortality on Reduviidae

(Hemiptera) at the concentrations we tested on aphids

(Cabral et al. 2000). Contact with lignan was also

responsible for important mortality on the hemipteran pest

Oncopeltus fasciatus (Cabral et al. 1999). The cytotoxic

activity reported for many lignans (Harmatha and Dinan

J. Saguez et al.

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2003) could be the mechanism that induced mortality in

aphids.

Several molecules (e.g., proteases inhibitors, lectins)

exert toxic effects by crossing the midgut epithelium and

affecting aphid demographic parameters and physiology

(Cherqui et al. 2003). Because lignans have been docu-

mented to affect insect growth, excretion, or enzymatic

metabolism (Bernard et al. 1989; Garcia and Azambuja

2004; Harmatha and Dinan 2003; Harmatha and Nawrot

2002; Nawrot et al. 1991; Russell et al. 1976), we suspect

that lignans can cross the midgut and have toxic effects in

hemolymph.

Aphids host obligatory endosymbiotic Proteobacteria,

Buchnera aphidicola. These endosymbionts produce nec-

essary amino acids for aphids. When Buchnera are elimi-

nated by chemical or antibiotic treatments, the resulting

aposymbiotic aphids are sterile and show reduced devel-

opmental performances and survival (Douglas 1992; Sasaki

et al. 1991). Lignans and neolignans are known to exert

antibacterial effects (MacRae and Towers 1984). They also

inhibit enzymatic systems (Bernard et al. 1989; MacRae

and Towers 1984) and can disrupt endosymbiont metabolic

functions. Although it was not demonstrated that lignans

directly act on Buchnera, mortality could be assigned to

such antibiotic and inhibitory effects of lignans and neo-

lignans leading to the death of obligatory endosymbionts

and ultimately affecting M. persicae survival and devel-

opment. In contrast, the absence of nymphal mortality at

50 lg/mL for DCG could reflect the ability of aphids to

compensate the lack of essential amino acids produced by

the endosymbionts. In fact, aphids experimentally lacking

endosymbionts develop a strategy which results in pro-

ducing high level of glutamine to partially compensate for

the lack of amino acids normally produced by Buchnera

(Wilkinson and Douglas 1995). We propose that at

10 lg/mL, DCG affects Buchnera but glutamine produc-

tion is not triggered, resulting in aphid mortality. At

50 lg/mL, aphids produce glutamine to compensate for the

lack of amino acids induced by the toxic effects of DCG on

Buchnera. At 100 lg/mL, this production of glutamine is

not sufficient to compensate for this lack. These mecha-

nisms could also explain the lack of effect of SECO on M.

persicae demographic parameters at 100 lg/mL.

Lignans and neolignans may also impact aphid growth

and fecundity by interfering with the hormonal system.

Aphid nymphal growth is punctuated by molts. Lignans have

been demonstrated to act as growth regulators, causing molt

disturbances due to antihormonal effects. As an example,

lignans display weak juvenile hormone activity (MacRae

and Towers 1984). Although the mode of action is unknown,

it was suggested that lignans disrupt the endocrine system of

insects and alter their hormones functions involved in

ecdysis and development (Harmatha and Dinan 2003). The

increase of PRP when aphids were reared on DCG and AHS

is the potential reflection of molting disturbance.

Lignans are phytoestrogens and SECO is the precursor

of enterodiol, a mammalian lignan obtained after hydro-

lysis by colonic bacteria in mammals. Enterodiol mimics

the effects of estrogens. However, some antiestrogenic

effects were also reported depending on the concentrations

(Dixon 2004). We have no evidence that SECO and other

lignans are metabolized in insect gut, but if this mechanism

exists, it could explain the different effects observed on

fecundity, notably with SECO and DCG where the DF was

significantly reduced by ca. 20–35 % at concentrations

lower than 100 lg/mL. Consequently, DCG and SECO

expressed more extensive deleterious effects on the rm

values of M. persicae than SDG and AHS.

Relation between compounds’ structure and biological

activity

Our results have shown that structurally related lignans had

distinct biological effects on aphids. It has already been

Table 1 Nymphal survival of M. persicae reared on artificial diets

containing various concentrations of lignans and neolignan

Lignan

concentrations

(lg/mL)

No aphids

tested

Survival

(%)

v2 df P

DCG

0 (control) 63 92.06

1 70 80.60 9.08 20 0.98

10 63 73.02 42.11 20 \0.00

50 62 91.94 0.61 20 1.00

100 57 61.40 45.45 20 \0.00

SDG

0 (control) 48 85.42

1 71 69.01 8.99 9 0.44

10 68 72.06 20.40 9 \0.00

50 68 66.17 48.80 9 \0.00

100 52 57.69 49.40 9 \0.00

AHS

0 (control) 71 64.79

1 74 62.16 1.70 10 1.00

10 72 62.5 4.54 9 0.87

50 74 60.81 0.63 11 1.00

100 73 64.38 2.02 9 0.99

SECO

0 (control) 74 66.22

1 65 56.92 9.80 9 0.36

10 63 69.84 2.97 9 0.97

50 56 64.29 0.94 10 1.00

100 67 65.67 3.52 11 0.98

Biological activities of lignans and neolignans

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suggested that the polarity of plant compounds is a pre-

dominant factor for their biological activities (Sugahara

et al. 2008). Although the mode of action of lignans and

neolignans is mostly unknown, several authors reported

that molecular structures and polarities of the compounds

confer the antifeedant activity or interact with specific

receptors involved in different physiological functions,

such as the endocrine system of insects (Harmatha and

Nawrot 2002).

Among the chemical groups that confer activity to the

lignans, it was shown that not only the type of lignan, their

aromatic moieties, hydroxyl or glycosyl groups but also

methoxy or methylenedioxy groups are associated with

antifeedant or larval growth inhibitory activities in non-

hemipteran pests (Harmatha and Nawrot 2002; Messiano

et al. 2008). The number of hydroxyl groups and the

removal of the glycoside moiety are important features

playing a role in the estrogenic activity (Apers et al. 2003;

Dixon 2004; Lehraiki et al. 2010) and can explain the

effects observed on aphid fecundity. The invoked reasons

are linked to the ability of compounds to cross the plasma

membrane of the cell or, alternatively, to bind to lipophilic

targets such as cellular receptors.

Deleterious effects due to glycosylated compounds were

observed on aphids and other insects. Glycoalkaloids from S.

tuberosum exert repulsive or toxic effects on some insects

such as the coleoptera Leptinotarsa decemlineata (Sinden

et al. 1980, 1986) and Agriotes obscurus (Jonasson and

Olsson 1994) or the leafhopper Empoasca fabae (Sanford

and Ladd 1992) and the aphid M. euphorbiae (Guntner et al.

2000) while they appear inefficient against M. persicae and

Aulacorthum solani (Flanders et al. 1992). Comparing the

effects of the different glycoalkaloids with their corre-

sponding aglycone and their glycosyl fractions (tetra-, tri, di-

or monosaccharidic) led to the conclusion that their biolog-

ical activity relies on the aglycone structure, but it is highly

Table 2 Demographic parameters of M. persicae that reached adulthood and reared on artificial diets containing various concentrations of

lignans and neolignan

Demographic parameters Lignan concentrations (lg/mL) F P

0 (control) 1 10 50 100

DCG

Number of aphids 58 54 46 57 35

PRP (days) 9.2 ± 1.7a 10 ± 1.8b 10.7 ± 1.5c 10.4 ± 1.9bc 10.9 ± 1.7c 8.0 \0.00

DF (nymph female-1 day-1) 0.65 ± 0.41a 0.48 ± 0.28b 0.52 ± 0.18ab 0.48 ± 0.28b 0.66 ± 0.44a 3.7 \0.00

rm (female female-1 day-1) 0.16 ± 0.06a 0.12 ± 0.06bc 0.13 ± 0.03c 0.12 ± 0.04bc 0.10 ± 0.04b 8.7 \0.00

DT (days) 5.9 ± 1.5a 6.6 ± 2.4bc 6.8 ± 1.5c 6.8 ± 2.4b 7.9 ± 3.3d 11.8 \0.00

SDG

Number of aphids 41 49 49 45 30

PRP (days) 11.8 ± 2.2a 11.8 ± 2.2a 11.8 ± 1.9a 11.8 ± 2.3a 11.6 ± 1.8a 0.1 0.99

DF (nymph female-1 day-1) 0.53 ± 0.26a 0.47 ± 0.30a 0.56 ± 0.18a 0.66 ± 0.29b 0.55 ± 0.14a 3.7 \0.00

rm (female female-1 day-1) 0.12 ± 0.03a 0.11 ± 0.05a 0.12 ± 0.04a 0.11 ± 0.03a 0.08 ± 0.04b 4.8 \0.00

DT (days) 7.9 ± 4.1a 8.3 ± 4.3a 6.7 ± 2.5a 7.2 ± 3.1a 13.2 ± 10.6b 8.5 \0.00

AHS

Number of aphids 46 46 45 45 47

PRP (days) 11.2 ± 2.0a 11.7 ± 1.7a 11.9 ± 2.0a 16.0 ± 1.8b 13.6 ± 1.8c 49.4 \0.00

DF (nymph female-1 day-1) 0.58 ± 0.25a 1.32 ± 0.31b 0.64 ± 0.13a 0.64 ± 0.31a 0.61 ± 0.19a 72.6 \0.00

rm (female female-1 day-1) 0.11 ± 0.04a 0.07 ± 0.04a 0.12 ± 0.04a 0.09 ± 0.02b 0.11 ± 0.02a 11.5 \0.00

DT (days) 7.1 ± 3.2a 7.3 ± 3.2a 7.3 ± 3.0a 8.5 ± 2.6a 6.8 ± 1.7a 1.7 0.14

SECO

Number of aphids 49 37 44 36 44

PRP (days) 11.4 ± 2.2a 11.6 ± 1.9a 11.5 ± 2.5a 11.4 ± 1.6a 11.6 ± 2.3a 0.1 0.97

DF (nymph female-1 day-1) 0.70 ± 0.31a 0.45 ± 0.17b 0.44 ± 0.17b 0.51 ± 0.14b 0.67 ± 0.28a 12.6 \0.00

rm (female female-1 day-1) 0.13 ± 0.04a 0.09 ± 0.03b 0.10 ± 0.04b 0.09 ± 0.03b 0.13 ± 0.04a 14.3 \0.00

DT (days) 6.0 ± 1.9a 8.7 ± 2.2b 8.5 ± 4.0b 8.9 ± 4.5b 6.0 ± 2.0a 10.3 \0.00

Acronyms are defined as: PRP prereproductive period, DF daily fecundity, rm intrinsic rate of increase, DT doubling time of population. Results

are given as: mean ± SD. Values in the same row followed by the same letter indicate that they do not differ significantly, according to Fisher’s

PLSD test (P = 0.05). F Fisher’s test value of one-way ANOVA analysis; P P value of the Fisher’s test

J. Saguez et al.

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modulated by the chemical structure of carbohydrates, the

number of carbohydrates groups linked to the aglycone, and

their stereochemical orientation (Guntner et al. 2000; Li et al.

2003, 2007; Rayburn et al. 1994; Roddick et al. 2001).

Saponins are also known to affect insect digestion, growth,

and development by displaying various effects ranging from

beneficial to toxic. These effects are attributed to their

structural variability (Harmatha 2000). Other molecules,

such as oligosaccharides, have been reported to possess

aphicidal activities such as growth inhibition and increase of

mortality (Bultel et al. 2009; Dussouy et al. 2012; Saguez

et al. 2008). As examples, chitosan, allosamidin, and other

molecules that mimic chitin structure composed of N-ace-

tylglucosamine induced deleterious effects on M. persicae

(Dussouy et al. 2012; Saguez et al. 2006). The biological

activity was supposed to be attributed to the number of

glucoside units and their type of linkage. Consequently, the

presence of glucoside units on DCG and SDG may be

responsible for the deleterious effects observed on aphids.

Our results show for the first time that lignans and

neolignans can affect the survival, the development, and

the demographic traits of the aphid M. persicae. The effects

can be attributed to the structure of the molecules. Gly-

cosylated units seem to have impacts on mortality, and

hydroxyl groups altered more specifically the other

demographic parameters. Consequently, in a strategy of

developing new compounds with aphicidal effects, more

attention should be focused on the structure of the com-

pounds, depending on the targeted developmental stage.

To clarify the mode of action of the lignans and neo-

lignans on aphids, several methods should be investigated.

For instance, the feeding behavior could be studied by

electropenetrography to evaluate the potential antifeedant

effects on whole flax plants or in vitro with isolated lign-

ans. Antibacterial experiments should be done to explore

the potential antibiotic effects on Buchnera endosymbionts.

This first study on the effects of lignans on aphids sug-

gests that there is potential in working on the interactions

between aphids and lignan derivatives. Although we tested

the effects of only four lignans and neolignans from flax,

hundreds of lignans appear to be good candidates in the

context of crop protection programs based on natural

resources. Some of them probably exert more deleterious

effects. Lignans are easily extractable and could be sprayed

alone or in combination with other pesticides because syn-

ergistic activities have been reported (Harmatha and Nawrot

2002). Before the potential use of these compounds as bio-

pesticides, it will be important to evaluate their effects on

other pests, beneficial insects, and animals. For crop pro-

tection, varietal selection of plants that naturally express

high amounts of lignans could be another way of research.

Attoumbre et al. (2010) recently developed lignan

antibodies for immunolocalization in flax seeds. Using

these antibodies, it will be possible to identify the target

organs of these compounds in the insect and confirm their

direct effects on aphids. This strategy could bring valuable

knowledge on the mode of action of lignans, which remains

largely unknown (Harmatha and Dinan 2003).

Acknowledgments The authors thank Charles Vincent and Noubar

J Bostanian (Agriculture and Agri-Food Canada Saint-Jean-sur-

Richelieu, Quebec) for their helpful advices and revisions on the

manuscript and the ‘‘Centre de Valorisation des Glucides et des

Produits Naturels’’ (Amiens, France) for providing L. usitatissimumseeds.

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