Egypt. J. Prot. Res. Inst. (2020), 3 (1): 130 - 137

Egyptian Journal of Plant Protection Research Institute

www.ejppri.eg.net

Laboratory evaluation of different host and Taxodium distichum ethanolic extract on viridula (: ) Rehab, M. El-Gendy Plant Protection Research Institute, Agricultural Research Center, Dokki, Giza, Egypt. ARTICLE INFO Abstract: Article History The different host plants and ethanolic extract of Received: 23/ 1/ 2020 Taxodium distichum were evaluated on mortality and Accepted: 20/ 2/2020 some biological parameters of green sting bug Nezara Keywords viridula (L.) (Hemiptera: Pentatomidae) a major pest of Host plants, Nezara some economic crops. The data revealed that the adult viridula, Taxodium survival on okra pod, cabbage and lettuce were 52.94, 30 distichum, longevity and 27.27% with longevity 9.56, 7.66 and 6.8 days, male and female. respectively. In addition, the insecticidal activity of ethanolic extract of T. distichum, it had a more potent on controlling N. viridula where LC50 and LC90 were 7.49 and 14.41% after 72h post-treatment. The ethanolic extract induced noted a decline in the longevity of male (4.42, 3.14) and female (5.57, 2.71) at 5% and 10% conc., respectively in comparison to control 14.71 and 22.57 day for male and female. Finally, T. distichum ethanolic extract is evidence that it is a good efficient for control. Introduction The green sting bug Nezara vulgaris (L.) as a diet (Gonzales and viridula (L.) (Hemiptera: Pentatomidae) Ferrero, 2008 and Silva et al., 2011). The is a serious economic polyphagous pest green sting bug reared on different seed to most of the crops in Egypt due to its and plant tissue combination as green stylet penetrate the plant tissues causing snap beans, raw-shelled peanuts, damage to all developmental stage of the immature radish and immature plant and so it is difficult to control. A pods caused shorter to duration few studies are known concerning period from egg to adult (Panizzi and feeding techniques of N. viridula , Saraiva, 1993; Noda and Kamano, 2002 although N. viridula biology and ecology and Gonzales and Ferrero, 2008). In are extensively recorded (Huang and Integrated Pest Management, the Toews, 2012). There was a limited resistances of plant studies are techniques attempt to improve the Menusan’s green take part in population reduction bean method in evaluating laboratory (Souza et al., 2013). rearing of green sting bug on Phaseolus

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The using increase of chemical The of T. distichum collected insecticides or alternate one chemical from El-Orman garden Giza, Egypt and product leads to more problems in then dried under vacuum at 30 oc until controlling sting bugs (Musser et al. and crashed into powder. About 500g of 2011) which aid in developing sting bug T. distichum powder was steeped at room resistance to chemical insecticides and temperature in ethanol for one week. The environment pollution (Hart and extract was then filtered, concentrated to Pimentel, 2002 and Sosa-Gomez and dryness in a rotary evaporator at 50 oc. Silva, 2010). Consequently, the 3.Bioassay: alternative uses of plant extracts as 3.1. The effect of different host plants: natural pesticides are more remarkable The adult of N. viridula fed on because they have toxic bioactive cabbage, Brassica oleracea compounds that low toxicity on (Brassicaceae); lettuce, Lactuca sativa mammalian. (Asteraceae); castor , Ricinus Taxodium distichum communis (Euphorbiaceae); okra leaves (Cupressaceae) exhibited antioxidant, and pods, Hibisicus esculentus antitumor, cytotoxic, antiviral, (Malvaceae) under laboratory conditions antibacterial and antifungal activities in glass jars. The food source was (Ibrahim et al., 2006 and Kusumoto et renewed daily. The survival percent and al., 2010) because of its enrichment in longevity of adult green sting bug were glycosides flavonoids, monoterpenes, determined. diterpenes, and sesquiterpenes. The 3.2. The effect of Taxodium distichum insecticidal activity of T. distichum ethanolic extract: extracts was less evaluated. Two concentrations of T. The present work aims to estimate distichum ethanolic extract 5 and10% survival activity observed for N. viridula were prepared with distal water. 5μL of reared on the different host plant. Also, each concentration were applied on the the toxic effect of T. distichum fruit tergites of each adult with the topical ethanol extract on some biological micro applicator and 5μL distal water for parameters of N. viridula adult and control then one pair of male and female determined its LC50 and LC90 were was put in glass jar lined with filter paper evaluated. and covered with organza and feeding Materials and Methods with okra pods according to (Costa et al., 1.Insect source: 1998). Seven replicates for each Adults of N. viridula were treatment were used to evaluate the obtained from a colony reared on fresh mortality percent and longevity of male green bean, Phaseolus vulgaris L. in the and female adults, the number of laboratory with fixed temperature of 27+ deposited eggs per female and 2 oC , 65+10% RH and12:12 (L:D) h. hatchability. All bioassays were photoperiod at Pest Physiology Research established at 27°C± 2°C, RH of 65%±10 Department, Plant Protection Research %, and photoperiod of 12 hours. The Institute. The ethanolic extracts of mortality data after 48, 72 and 96h were Taxodium distichum were dissolved in corrected according to Abbott's formula ethanol. (1925). 2. Extraction of Taxodium distichum: 4.Statistical analysis:

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Egypt. J. Plant Prot. Res. Inst. (2020), 3 (1): 130- 137

Differences among mean were pods, H. esculentus on survival percent of analysed using Duncan's analysis of N. viridula, green sting bug. All host variance (ANOVA), the least significant plants caused 100% survival on the first difference by computer statistical day after feeding except castor leaves software Costat® (2005). The caused 37.3%. While on a ninth day the significance test at probability value p < adult survival on okra pod, cabbage and 0.05 was considered significant. The lettuce were 52.94, 30 and 27.27%, toxicity line of T. distichum ethanolic respectively. In the same trend, regarding extract were analyzed with Biostat Table (1) the longevity of adult green version 5 using Probit-analysis. sting bug was 9.56, 7.66 and 6.8 days on Results and discussion okra pods, cabbage and lettuce host 1.Effect of different host plants on plants on the contrary, the lowest adult survival: longevity of adult 2.5 days was recorded Data in Figure (1) illustrated the on castor leaves. There were highly effect of different host plants as cabbage, significant differences in the longevity of B. oleracea; lettuce, L. sativa; castor N. viridula adult feeding on the various leaves, R. communis; okra leaves and host plants.

Figure (1): Survival percentage of adult Nezara viridula reared on different host plants.

Table (1): Effect of different host plants on Nezara viridula adult longevity.

Host plants Longevity (days) Cabbage leaves 7.66b+0.12 Lettuce leaves 6.8bc+0.15 Castor leaves 2.5d+0.28 Okra leaves 5.53c+0.28 Okra pods 9.56a+0.79 LSD 1.28 P 0.0000*** L.S.D. means low significance differences at P< 0.05 2. Effect of Taxodium distichum mortality percent of adult green sting bug ethanolic extract on mortality of at 5% concentration was slightly Nezara viridula adult: decreased after 48h (7.14) then increased The efficacy of two concentrations after 96h reached to 50%. On the other (5 and 10%) T. distichum ethanolic hand, the second concentration 10% extract were observed in Table (2). The exhibited more toxic after 48, 72 and 96h

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El-Gendy, 2020 and the mortality % were 35.71, 71.43 adults increased with increasing and 100%, respectively on comparison to concentrations of T. distichum as well as control. The mortality of green sting bug experimental duration. Table (2): Mortality % of Nezara viridula adult infected with ethanolic extracts of Taxodium distichum under laboratory conditions. Treatment Conc. Mortality% (%) 48h 72h 96h Taxodium distichum 5% 7.14 21.42 50 10% 35.71 71.43 100 Control 0.00 0.00 0.00

3.Susceptibility of Nezara viridula adult and 90%; LC50 and LC90 mortality to to Taxodium distichum ethanolic green sting bug adults were 7.49 and extract: 14.41% with slope 4.50 after 72h post- Regarding Table (3) and Figure treatment. The mortality of N. viridula (2) the lethal concentration of T. increased with increasing concentrations distichum ethanolic extract caused 50% of T. distichum ethanolic extract. Table (3): The lethal concentrations of Taxodium distichum ethanolic extract against Nezara viridula. Ethanolic Lethal concentration Lower limit Upper limit slope extract of (%) Taxodium LC 7.49 5.29 11.64 50 4.50 distichum LC90 14.41 10.06 18.27

Figure (2): Probit-analysis of toxicity lines of Taxodium distichum ethanolic extract on Nezara viridula adult. 4.Effect of Taxodium distichum 3.14 and 14.71day at 5%, 10% and ethanolic extract concentrations on control, respectively. In the same context, some biological parameters of Nezara the female longevity was 5.57, 2.71 and viridula adult: 22.57 days at 5%, 10% and control, The obtained results in Table (4) respectively. There was no egg deposited and Figure (3) illustrated the longevity of at 10 % concentration subsequently, there male and female, mean no. of deposited was no hatchability. While at 5% there egg per female and hatchability% of were 11.28 eggs with 7.49% hatchability green sting bug. There were highly in comparison with control was 55.71 significance differences between all eggs with 93.27% hatched. In conclusion, treatments in various observed biological all tested biological parameters were parameters. The male longevity was 4.42, highly decreased than the control.

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Egypt. J. Plant Prot. Res. Inst. (2020), 3 (1): 130- 137

Table (4): Biological parameters of Nezara viridula adult after treatments with 5&10% Taxodium distichum ethanolic extract. Longevity(days) Mean no of Mean+SE deposited Hatchability% Treatments Egg/female Mean +SE Male Female Mean+SE 5% 4.42b+0.29 5.57b+0.61 11.28b+4.13 7.49b+3.08 10% 3.14b+0.34 2.71c+ 0.28 0.00c+0.00 0.00c+0.00

Control 14.71a+0.91 22.57a+0.84 55.71a+2.73 93.27a+1.39

LSD 1.75 1.85 8.50 5.8

P 0.0000*** 0.0000*** 0.0000*** 0.0000*** Same letters mean non-significant effect Different letters mean significant effect at p<0.05.

A B C

Figure (3): Effect of ethanolic extract of Taxodium distichum on egg deposit. A. Mating of male and female after treatment. B. Normal deposit egg in control. C. Unviable egg in treatment with 5% conc. The different host plants under mortality of green sting bug reared on the investigation are effects on survival artificial diet and soybean pods was 30%. percent of N. viridula, green sting bug. In the same trend, the adult of green sting The survivorship of adult on okra pod, bug survival was 97.3 and 74.67% on cabbage and lettuce were 52.94, 30 and fresh and dry yolk chicken egg-based 27.27% and the longevity of adult green diet, respectively (Portilla et al., 2015). sting bug were 9.56, 7.66 and 6.8 days, Also, the duration of N. viridula reared respectively on contrary the castor leaves on immature soybean pods was 90.6 d B not a preferred host plant to of adult 2.5 (Gonzales and Ferrero, 2008). days was recorded on castor leaves. The concern of rearing techniques There were highly significant differences of green sting bug on different host plants in the longevity of N. viridula adult is not only to investigate the population feeding on various host plants. These dynamics in the laboratory but also to results are in harmony with (Panizzi et design its control strategies. The obtained al., 2000) stated that the low nymphal results indicated that the ethanolic extract

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El-Gendy, 2020 of T. distichum has insecticidal activity extract which responsible for its toxicity. on N. viridula adult where LC50 and LC90 (Sabry, 2018) recorded that Ferruginol, were 7.49& 14.41% with slope 4.50 after (Di-(2-ethylhexyl) phthalate), piperine, 72h post-treatment. These results are in 3alpha, Didecyl phthalate and the same context as the results of (Sabry, octadecane, 1-[2-(hexadecyloxy) ethoxy] 2018) revealed that the LC50 and LC90 are the major active compounds. were 10490 ppm and 27890 ppm with Furthermore, the existence of taxodione slope 3.007 on 4th instar larvae of compound; a quinone methide diterpene Spodoptera littoralis after 72 hours of that possesses insecticidal activity and treatment. The contact and the stomach DNA-binding effects (Fraga et al., 2005 poison activity of petroleum ether and and Zaghloul et al., 2008). Besides, it acetone extracts of leaves from bald acts as cholinesterase inhibitor (Kuźma et cypress (T. distichum) were tested on al., 2016 and Liu et al., 2014) and adults Tribolium castaneum Herbst, cytotoxic activities, causing apoptosis in mortality percentage increased with several tumor cell lines as well as concentration and exposure time Phthalate compound is a more efficient as increasing (Shoukry et al., 2017). larvicidal activity Khatiwora et al. The ethanolic extract of T. (2013). distichum fruits caused highly decreased It is concluded that the okra pod in all tested biological parameters; plant can be used in rearing techniques of longevity, deposited egg/female and green sting bug under laboratory hatchability than the control, there was no conditions. The castor leaves not a egg deposited at 10 % concentration preferred host plant to N. viridula so it while at 5% there was 11.28 egg with can be cultivated surrounding the 7.49% hatchability in comparison with economic crop. The insecticidal activity control was 55.71 egg with 93.27% and the latent effect of the biological hatched. These results are supported by parameter of N. viridula adult may be Piton et al. (2014) evaluate the contact regarding the active chemical compounds toxicity of the leaves acetonic extract of of T. distichum ethanolic extract. These Piper aduncum (L.) on brown stink bug, promising results are encouraged to Euschistus heros developmental stages, schedule T. distichum ethanolic extract all concentrations tested reduced into insect management programs. significantly the survival and References reproduction in the adult bioassay. In the Abbott, W. S. (1925): Methods for effect on egg contact bioassay the 8% computing the effectiveness of concentration caused 19% mortality. In insecticide. J. Econ. Ent., 18(2): the same frame of reference Carneiro et 256-267. al. (2013) found that ovicidal action of Carneiro, Â. P.; Pereira, M. J. B. and extract on the chorion of Galbiati, C. (2013): Biocide Rhodnius neglectus (Lent.) (Hemiptera: activity of coriacea seeds Reduviidae) eggs caused unviability of extract on Rhodnius neglectus 90% of the eggs. (Hemiptera: Reduviidae). Revista The mortality, low longevity and de Biologia tropical, 61(1): 419- fertility of N. viridula adults may be 427. related to the presence of the bioactive Costa, M. L.; Borges, M. and Vilela, E. compounds in T. distichum ethanolic F. (1998) : Biologia reprodutiva de

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