R E S E A R C H A R T I C L E
The Effect of Photoperiods on the Insecticidal Activity
of Hypericum perforatum Extract on the Third Larval Instar of Diamondback Moth, Plutella xylostella
Milad Ebrahimi Fakhar1• Jaber Karimi1•Alireza Rezazadeh1• Habib Abbasipour1 • Amir Mohammad Naji2•Alireza Askarianzadeh1
Received: 16 September 2021 / Revised: 15 March 2022 / Accepted: 20 March 2022 ÓZoological Society, Kolkata, India 2022
Abstract Insecticidal activity of Hypericum perforatum extract towards third larval instars of the diamondback moth, Plutella xylostella in different concentrations including 5, 100, 1000, and 5000ll/ml was tested at dif- ferent photoperiods (12L:12D, 16L:8D, 8L:16D, 20L:4D and 4L:20D hours). The obtained values of LC10, LC25, and LC50 in normal photoperiod were 11, 74, and 622ll/
ml, respectively. The results showed a direct relation between insecticidal activity of hypericin and its concen- tration. Thus, values of LC10, LC25, and LC50 provided poor, fair, and good control, respectively. When these concentrations were tested in different photoperiods, results showed that the mortality of the test animals increased with increasing photoperiod in constant concen- tration. The mortality varied depending on light hours in photoperiods, with more light caused higher mortality.
Results showed that the extract with a low concentration of
5 ll/ml was also effective in feeding indices of P.
xylostella.
Keywords Plutella xylostellaHypericum perforatum Plant extractInsecticidal activityPhotoperiods
Introduction
The diamondback moth,Plutella xylostellaL. (DBM) (Lepidoptera: Plutellidae), is among the most important pests of brassicas grown around the world (Soufbaf et al.
2012; Furlong et al.2013). A conservative estimate of total costs associated with diamondback moth management, thus, US$ 4–5 billions (Zalucki et al.2012) and, in Iran, it is about US$ 4,736,970 based on the weekly application of insecticide (FP) (FAOSTAT 2012). Plutella xylostella is one of the most resistant species of insects (Mota-Sanchez et al.2002). Success of DBM as a major pest is attributed to its ability to survive under a wide temperature range, prolific reproductive capacity, ability to feed on diverse host plants, and insecticide resistance (Vickers et al.2004;
Shelton2004). Over the last two decades, DBM has been a major pest of cruciferous crops in Tehran province and other areas of Iran (Mahmoudvand et al.2009). Owing to its ability to develop resistance to many conventional insecticides, the use of new insecticides that have low effect on other non-target organisms can be effective and helpful. New natural plant-based pesticides can help as these act specifically as inhibitors of oviposition and feeding of pests and have no adverse effects on non-target organisms (Miresmailli and Isman2014). Unlike synthetic materials, phytochemicals are complex substances that can delay the resistance of pests to insecticides (Isman et al.
1990; Velasques et al.2017).
& Habib Abbasipour
[email protected] Milad Ebrahimi Fakhar [email protected] Jaber Karimi
[email protected] Alireza Rezazadeh [email protected] Amir Mohammad Naji [email protected] Alireza Askarianzadeh [email protected]
1 Department of Plant Protection, Faculty of Agriculture, Shahed University, Tehran, Iran
2 Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahed University, Tehran, Iran
Proc Zool Soc
https://doi.org/10.1007/s12595-022-00440-7
TH TY
K O LK ATA
The use of photochemical processes as a tool to control the population of different insects has been repeatedly examined in both laboratory experiments and field studies (Luksˇien_e et al.2007; Cavoski et al. 2011). Most investi- gations have been performed by using photoactivat- able polycyclic aromatic dyes that absorb near-UV light wavelengths, including thiophenes, furocoumarines, and quinones (Ben Amor and Jori2000). Along the same line, Ben Amor et al. (1998) demonstrated that haematopor- phyrin is an efficient phototoxic to several insects including Ceratitis capitata (Wiedemann) and Bactrocera oleae (Rossi) (Diptera: Tephritidae). Hypericum spp., from Hypericaceae family includes nearly 484 species (Guedes et al.2012).Hypericum perforatumL., (common St John&s wort) has been included in the pharmacopeia of many countries (Ivanova et al. 2005; Jaric´ et al. 2007). The insecticidal effects of essential oils from someHypericum spp., have been assessed in previous studies (Kordali et al.
2012; Rouis et al. 2013) and these plant materials have been candidates for future work in the management of pests. Many species from the genusHypericumcontain the phototoxin hypericin (4, 5, 7, 40, 50, 70-hexahydroxy-2, 20- dimethylnaphtodiant rone), which is located in dark glands in the leaf, stem, and flower. In addition to glands con- taining hypericin, Hypericum perforatum L. has clear glands that may transmit large amounts of visible light through the leaf (Jendzˇelovska´ et al.2016). The insecticidal effects ofHypericumspp. have been assessed againstCulex pipiens (L.) (Diptera: Culicidae) (Rouis et al. 2013),Tri- bolium castaneum (Herbst) (Coleptera: Tenebrionidae) (Parchin and Ebadollahi 2016),Manduca sexta L. (Lepi- doptera: Sphingidae) (Samuels and Knox1989) and Sito- philus granarius(L.) (Coleptera: Dryophthoridae) (Kordali et al.2012).
The objective of the present study was to address some of these issues.
Materials and Methods Extraction ofH. perforatum
Thirty grams of cut and dried plant material (leaves and flowering branches) was extracted using 1000 ml of ultra- distilled water (15 min, 85°C, 3000 rpm). The herb par- ticles were then separated from the solution using a 10lm filter. The separated solution was put in a vacuum rotary evaporator at 60°C for 1 h to remove water content and the resultant extract was then dried for 24 h at 45°C in a vacuum concentrator (Eppendorf, Germany) (Barnes et al.
2001). In experiments, the extracts were dissolved in Milli- Q water with a concentration of 10 g L-1. The obtained extract was poured into dark containers in a package and
stored in a refrigerator at 4°C. This served as the stock material and it was used to prepare solutions and subse- quent concentrations. Due to photodynamic nature of compounds, extract preparations were used within a short time.
Insect Rearing
The initial P. xylostella colony was collected from cauli- flower fields of Shahre-Rey in south of Tehran, Iran. For egg-laying, about 500 adults ofP. xylostellawere placed in a plastic cage (50930930 cm) and eggs were trans- ferred to leaves of cauliflower, Brassica oleracea var.
botrytis as food material to continue their development.
Insect stock was maintained at 25±1°C and 65% ±5%
relative humidity under a 16:8 (L:D) hour photoperiod in a growth chamber.
Bioassay ofH. perforatum Extract on 3rd Instar Larvae ofP. xylostella
For bioassay experiment on the insecticidal properties of the extract ofH. perforatum, the same procedure as above was used (Tabashnik and Cushing 1987). Six concentra- tions, ranging between 5 to 95% mortality of extract were determined by preliminary tests. Cabbage leaf disks (3 cm in diameter) were dipped in different concentrations (in- cluding 25, 50, 500, 1000, 5000 and 10,000ll/ml) of extract solutions containing 0.02% Tween-80 for 30 s. For the control, the leaf disks were dipped in water with 0.02%
Tween-80. The treated leaf disks were permitted to dry at room temperature for 30 min. Then those were used in treatments. Four replicates and at least 60 third instars were used for each concentration. Mortality was recorded 24 h after treatment.
Effect of Photoperiod Duration on Mortality byH.
perforatumExtract
In the extract mortality test, different light periods (12L:12D, 16L:8D, 8L:16D, 20L:4D, and 4L:20D hour) were tested. The mortality rates at LC10, LC25, and LC50 concentrations on 3rd instar larvae of P. xylostella were recorded in five different photoperiods. The day before the test, temperature, humidity, and light period of the germi- nators were adjusted. In all experiments, temperature and relative humidity were kept constant and the light period varied according to the experiment. All the experiments were carried out to study the effect of photoperiod on the lethality of H. perforatum extract at 27±2°C and 65±5% RH.
Effect ofH. perforatum Extract on Nutritional Indices ofP. xylostella
Nutritional indices were determined following the method described by Farrar et al. (1989) with some modifications by Dastranj et al. (2018). The feeding deterrence index (FDI) was calculated according to Isman et al. (1990).
Initially, larvae were reared on the treated and control plants until attainment of 10-day age of third instar. Four groups of 10 one-day-old larvae each were prepared, weighed, and transferred into glass Petri dishes (12 cm diameter and 1.5 cm depth) containing the fresh B. oler- aceavar.botrytisleaf of each treatment or control. Petioles of the leaf disks were inserted into cotton ball soaked in water to maintain freshness and these were replaced daily with new one. For 4 days, the initial fresh food, food remnant, and feces remaining at the end of each experiment were weighed in an electric balance (Model#SE-391).
Also, the larvae in each paired Petri dishes were checked daily for mortality or ecdysis. Nutritional indices were calculated as under:
1. Relative growth rate (RGR) = Insect wt. gain/Insect wt. at beginning of treatment (mg)9time (day) 2. Relative consumption rate (RCR) = Wt. food eaten/
Insect wt. at beginning of treatment (mg)9time (day) 3. Efficiency of conversion of ingested food (ECI) = [(in-
sect wt. gain)/(wt. food eaten)]9100
4. Feeding Deterrence Index (FDI %) = [(C—T)/
C]9100, where C = the food consumption in control leaves, T = food consumption in treated leaves, as the control and treated leaves were placed in separate vials in no-choice tests
Statistical Analysis
The data from the experiments were subjected to one-way analysis of variance (ANOVA) (P\0.05), Kruskal–Wallis and Jonckheere-Terpstra tests. Mean values were compared by Tukey Multiple Range Test. Values of LC10, LC25, and LC50with 95% fiducial limits (FL) were calculated, using Probit analysis. A significance level of P\0.05 was considered to reject the null hypothesis. SAS 9.2 software (SAS Institute2009) was used for the analysis of data.
Results
Bioassay ofH. perforatumExtract on 3rd Instar Larvae ofP. xylostella
The results showed that with increasing the concentration of the extract, the mortality rate also increases. Thus, LC10
concentration with 11ll/ml, LC25with 74ll/ml, and LC50
with 622ll/ml produced the lowest (11.32%) and highest (57.3%) mortality of test animals, respectively (Fig.1).
There was a significant difference in mortality between the three concentrations.
Effect of Photoperiod Duration on Mortality byH.
perforatumExtract
The Fig.2 show LC10, LC25, and LC50values at different concentrations of H. perforatum extract in different pho- toperiods. These values are significantly different from each other. Mortality increased at longer light hours. The 20L: 4D h light regime registered the highest mortality of 13.30%, 30%, and 60.56%, respectively. The 4L: 20D h light regime showed the lowest mortality of 0, 10 and 33.30%, respectively (Fig.2).
Effect ofH. perforatumExtract on Nutritional Indices ofP. xylostella
The effect ofH. perforatumextract on feeding indices ofP.
xylostellais provided in Table1. No significant difference occurred in RGR at different concentrations and in the control. But, RCR reduced significantly at concentrations of 3ll/ml and 5ll/ml than that recorded at other con- centrations. A significant difference occurred among treatments in the ECI index and FDI of different treatments and control. FDI increased and ECI decreased at increasing concentrations of hypericum plant extract (Table 1).
Results clearly showed that a concentration of 0.5ll/ml caused significantly higher feeding deterrence (FDI) of leaf disks than other concentrations.
Discussion
Results showed significant influence of light periods in the efficacy ofH. perforatumextract on 3rdinstar larvae of the diamondback moth, P. xylostealla, Effect of the amount light emitted in each photoperiods on the mortality rate of the larvae differed in different light periods. The mean mortality recorded at LC10 concentration in 4D: 20L pho- toperiod is found to be higher than the mean mortality recorded at LC25 concentration in 20D: 4L photoperiod, although the concentration of the extract in LC25is about 7 times the concentration of the extract in LC10. The mean mortality rate recorded at LC25 concentration in 4D: 20L photoperiod was nearly similar to the average mortality recorded at LC50 concentration in 20D: 4L photoperiod.
This was despite a significantly higher concentration of the extract in LC50(622 ll/ml) in comparison to that of LC25 (74ll/ml). These results clearly suggested that the light
seems to work in synergy with the plant extract, and between these light intensity appears to contribute signifi- cantly more in enhancing the efficacy of the plant extract on the test organism of this study.
An earlier study recorded LC50 at a concentration of 21,731 ppm ofH. perforatum extract against 3rdinstar of P. xylostella(Askarianzadeh et al. 2012). Other previous studies using essential oil ofH. perforatum have demon- strated toxicity to Tenebrio molitor L. and Triblium cas- taneum (Herbst) (Bas¸ and Ersoy 2020; Parchin and Ebadollahi2016).
In present study, results showed that H. perforatum extract has negative effect on nutritional indices and a
strong feeding deterrence effect on the larvae ofP. xylos- tella. Charleston et al. (2006) reported that plant leaf extract ofMelia azedarachL. andAzedirachta indicahave negative effects on the survival, regeneration, growth, oviposition, and feeding indices of diamondback moth.
Kostic´ et al. (2008) reported larvicidal and anti-feeding effects of ethanolic solution of essential oils of Ocimum basilicum L. on the gypsy moth, Lymantria dispar (L.).
Extracts ofNerim oleanderL.,Lavandula officinalis Mill.
and Ferula assa-foetida L. have anti-feeding effect on Tribolium castaneum(Herbst) (Moharamipour et al.2001).
Fig. 1 The concentrations ofH.
perforatumextract responsible of lethal (LC50) and sublethal (LC10and LC25) effect on the 3rd instar larvae ofP. xylostella under laboratory condition
Fig. 2 Comparison of the mortality recorded at LC10, LC25and LC50ofH. perforatum extract on the 3rd instar larvae ofP. xylostellain different photoperiods
Conclusion
The current results showed thatH. perforatumextracts had phototoxin activity on the diamondback moth,P. xylostella larvae varying from nearly 11% to 57% mortality. The main advantage of sunlight-activatable photoinsecticides is certainly represented by their lack of toxicity towards most biological systems in the dark, which minimizes their impact on the environment. However, some concerns still exist especially as regards the induction of phytotoxicity.
Acknowledgements The work received financial support from the Postgraduate Education Bureau of Shahed University, Iran, which is greatly appreciated.
Declarations
Conflict of interest The authors declare that they have no conflict of interest.
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Table 1 Mean±SE values of nutritional indices ofP. xylostellaat different concentrations ofH. perforatumextract
Concentrations of extract (ll/ml) FDI% ECI% RCR (mg/day) RGR (mg/day)
0 0.00±0.00a 5.11±0.12a 198.84±11.81ab 10.29±0.85
0.5 9.61±1.00ab 4.95±0.35a 248.79±88.15a 16.89±5.61
1.5 42.81±7.73b 5.79±0.29ab 135.28±15.18ab 7.88±1.13
3 86.86±8.81c 6.52±0.08c 93.22±11.11b 6.07±0.69
5 24.86 c±91.12 5.84±0.57ab 79.61±7.71b 4.58±0.42
F (df) (4) 17.882** (4) 3.338* (4) 3.071* (4) 1.679 ns
P 0.0001 0.038 0.049 0.207
**,*Significant at 1%, 5%; respectively,nsdenote Not significant
Different small letters accompanying mean values in each column denote significant differences by Tukey’s HSD
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