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Insecticidal activity of Vitex mollis
Fitoterapia 78 (2007) 37 – 39 www.elsevier.com/locate/fitote
Short report
Insecticidal activity of Vitex mollis V. Rodríguez-López a,⁎, M.Z. Figueroa-Suárez a , T. Rodríguez b , E. Aranda c a
Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico b Centro de Estudios Ambientales e Investigación Sierra de Huautla, Universidad Autónoma del Estado de Morelos, Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico c Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos (UAEM), Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico Received 22 October 2005; accepted 6 September 2006 Available online 23 September 2006
Abstract Organic extracts from the leaves of Vitex mollis were assessed for their toxic effect on fall armyworm neonate larvae (Spodoptera frugiperda), an important insect pest of corn. The extracts showed insecticidal and insect growth regulatory activity, being CHCl3– MeOH (1:1) extract the most active, also found to be toxic in the Artemia salina test. © 2006 Published by Elsevier B.V. Keywords: Spodoptera frugiperda; Vitex mollis; Artemia salina; Insecticidal activity
1. Plant Vitex mollis Kunth (Verbenaceae) [1], leaves collected at Pitzotlan, Tepalcingo, Morelos, Mexico, in November 2002. Voucher samples (HUMO-19052) were deposited at Centro de Educación Ambiental e Investigación Sierra Huautla Herbarium (HUMO). 2. Uses in traditional medicine V. mollis is reported as a remedy to alleviate dysentery, as well as an analgesic and anti-inflammatory medicine [2]; other folk uses include treatment of scorpion stings, diarrhea and stomach ache [1]. 3. Previously isolated constituents Soluble and insoluble dietary fiber, tannin, copper, chromium, zinc, phytic acid and nickel [3].
⁎ Corresponding author. E-mail address: [email protected] (V. Rodríguez-López). 0367-326X/$ - see front matter © 2006 Published by Elsevier B.V. doi:10.1016/j.fitote.2006.09.020
38
V. Rodríguez-López et al. / Fitoterapia 78 (2007) 37–39
Table 1 Mortality, LC50, weight and days to pupation of S. frugiperda larvae feed on a meridic diet supplemented with leaf extracts of Vitex mollis Extracts
μg/ml
Weight gained (mg)
CH2Cl2
25 75 125 5 25 50 75 25 75 125
16.56 ± 0.50 14.52 ± 0.45 12.66 ± 0.18 10.29 ± 0.20 11.56 ± 0.16 10.65 ± 0.11
82.47 72.31 63.04 51.24 57.56 53.03
14.52 ± 0.45 12.72 ± 0.21 10.97 ± 0.13
73.31 63.34 54.63
CHCl3–MeOH (1:1)
MeOH
Fosdrim a Control a
20.08 ± 0.33
% of weight
100
% mortality
LC50
41.20 52.00 72.20 77.25 88.26 97.89 100 30.91 56.91 64.33 100 0
46.35
13.63
61.05
6.41
Positive control (25, 75 and 125 ppm).
4. Tested material Methylene chloride (CH2Cl2), chloroform–methanol (CHCl3–MeOH 1:1) extracts. 5. Studied activity Insecticidal properties and insect growth regulatory effect against Spodoptera frugiperda by direct contact method [4], cytotoxicity by brine shrimp lethality test (BSLT) [5]. Criteria of evaluation. Larval mortality and LC50; larval weight, the growth index (GI), relative growth index (RGI), duration of pupal stage, and successfully pupation of S. frugiperda larvae [4] and LC50 of Artemia salina [5]. Data were analyzed by ANOVA [6], where results are given as probability values, with P b 0.05 adopted as the criterion of significance. Lethality was analyzed with a Finney computer program (Probit Analysis) to determine LC50. 6. Used organism S. frugiperda larvae used for the experiments were obtained from culture at the Centro de Investigación en Biotecnología at the Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico, maintained under previously described conditions [4].
Table 2 Effect of V. mollis leaf extracts on growth index (GI), relative growth index (RGI), duration of pupal stage, and successfully pupation of S. frugiperda and LC50 of A. salina Extracts
μg/ml
GI
RGI
Duration of pupal stage
Successful pupation %
BSLT a LC50
Control CH2Cl2
– 25 75 125 5 25 50 25 75 125
0.95 0.58 0.47 0.27 0.25 0.12 0.04 0.68 0.43 0.35
1.0 0.59 0.48 0.27 0.25 0.12 0.04 0.69 0.44 0.36
15.9 ± 0.16 13.2 ± 0.17 14.6 ± 0.25 13.8 ± 0.23 15.9 ± 0.28 13.6 ± 0.16 14.8 ± 0.40 13.9 ± 0.36 14.7 ± 0.41 12.3 ± 0.34
98 83 91 86 47 66 – 87 92 77
– 358.26
CHCl3–MeOH (1:1)
MeOH
a
BSLT, Brine Shrimp Lethality Test.
247.10
122.92
V. Rodríguez-López et al. / Fitoterapia 78 (2007) 37–39
39
7. Results Reported in Tables 1 and 2. 8. Conclusion This study could contribute to the assessment of the possibility of using medicinal plants as potential insecticides [7]. The extracts were evaluated for their effect on the fall armyworm (S. frugiperda), an important insect pest of corn. All the extracts showed insecticidal and insect growth regulatory activity. The most active extract was CHCl3–MeOH (1:1) with LC50 13.63 ppm and 100% larval mortality at 75 ppm. On the other hand, this extract showed an important antifeeding activity against S. frugiperda (differences in larval weights were found between the control group and treatments group). Concerning insect growth inhibitory activity, the percentage of larvae that reached pupation decreased drastically with almost all extracts; larvae that reached pupal stage belonged to the lowest concentration group. The CHCl3– MeOH (1:1) extract had a 47% delay of pupation at 5 ppm, which was significant when compared with the data reported in the literature for methanolic extracts prepared from Yucca periculosa (25 ppm, 37.5%) used as the positive control [4]. Additionally, the extracts showed an important activity when tested to determine their toxicity to A. salina [5]. The results obtained have provided evidence to the potential use of V. mollis as insecticide. Bioassay-directed fractioning of the most active crude extracts to isolate and identify responsible compounds of the insecticidal activity and their possible mechanism of action is in progress. Acknowledgements The authors thank Biologist Laura Patricia Lina García and Biologist Mayra Pichardo for their valuable help in the biological assays. This study was financed by Grants from PROMEP-SEP and CONACyT (Convenio SEP-2003-CO243440/280). M. Z. Figueroa-Suárez acknowledges a scholarship (186641) from CONACyT. References [1] [2] [3] [4] [5]
Monroy O, Castillo E. Plantas medicinales utilizadas en el Estado de Morelos. Cuernavaca, México: CIB-UAEM; 2000. p. 260. Argueta A, Cano LM, Rodarte ME. Atlas de las plantas de la medicina tradicional, vol. I and III. México: Instituto Nacional Indigenista; 1994. Montiel-Herrera M, Camacho-Hernández IL, Ríos-Morgan A, Delgado-Vargas F. J Food Compos Anal 2004;17:205. Céspedes CL, Salazar JR, Martínez M, Aranda E. Phytochemistry 2005;66:2481. McLauglhlin JM. Crow gall tumors on potato disc and brine shrimp lethality: two simple bioassays for higher plant screening and fractionation. In: Hostettmann K, editor. Assays for bioactivity. San Diego: Academic Press; 1991. [6] Keppel G. Design and analysis: a researcher's handbook. New York: Prentice Hall; 1982. [7] Bowers SW. Insecticidal compounds from plants. In: Nigg HN, Seigler D, editors. Phytochemical resources for medicine and agriculture. New York: Plenum Publishing; 1992.
Short report
Insecticidal activity of Vitex mollis V. Rodríguez-López a,⁎, M.Z. Figueroa-Suárez a , T. Rodríguez b , E. Aranda c a
Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico b Centro de Estudios Ambientales e Investigación Sierra de Huautla, Universidad Autónoma del Estado de Morelos, Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico c Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos (UAEM), Avenida Universidad 1001, Colonia Chamilpa, Cuernavaca Morelos 6209, Mexico Received 22 October 2005; accepted 6 September 2006 Available online 23 September 2006
Abstract Organic extracts from the leaves of Vitex mollis were assessed for their toxic effect on fall armyworm neonate larvae (Spodoptera frugiperda), an important insect pest of corn. The extracts showed insecticidal and insect growth regulatory activity, being CHCl3– MeOH (1:1) extract the most active, also found to be toxic in the Artemia salina test. © 2006 Published by Elsevier B.V. Keywords: Spodoptera frugiperda; Vitex mollis; Artemia salina; Insecticidal activity
1. Plant Vitex mollis Kunth (Verbenaceae) [1], leaves collected at Pitzotlan, Tepalcingo, Morelos, Mexico, in November 2002. Voucher samples (HUMO-19052) were deposited at Centro de Educación Ambiental e Investigación Sierra Huautla Herbarium (HUMO). 2. Uses in traditional medicine V. mollis is reported as a remedy to alleviate dysentery, as well as an analgesic and anti-inflammatory medicine [2]; other folk uses include treatment of scorpion stings, diarrhea and stomach ache [1]. 3. Previously isolated constituents Soluble and insoluble dietary fiber, tannin, copper, chromium, zinc, phytic acid and nickel [3].
⁎ Corresponding author. E-mail address: [email protected] (V. Rodríguez-López). 0367-326X/$ - see front matter © 2006 Published by Elsevier B.V. doi:10.1016/j.fitote.2006.09.020
38
V. Rodríguez-López et al. / Fitoterapia 78 (2007) 37–39
Table 1 Mortality, LC50, weight and days to pupation of S. frugiperda larvae feed on a meridic diet supplemented with leaf extracts of Vitex mollis Extracts
μg/ml
Weight gained (mg)
CH2Cl2
25 75 125 5 25 50 75 25 75 125
16.56 ± 0.50 14.52 ± 0.45 12.66 ± 0.18 10.29 ± 0.20 11.56 ± 0.16 10.65 ± 0.11
82.47 72.31 63.04 51.24 57.56 53.03
14.52 ± 0.45 12.72 ± 0.21 10.97 ± 0.13
73.31 63.34 54.63
CHCl3–MeOH (1:1)
MeOH
Fosdrim a Control a
20.08 ± 0.33
% of weight
100
% mortality
LC50
41.20 52.00 72.20 77.25 88.26 97.89 100 30.91 56.91 64.33 100 0
46.35
13.63
61.05
6.41
Positive control (25, 75 and 125 ppm).
4. Tested material Methylene chloride (CH2Cl2), chloroform–methanol (CHCl3–MeOH 1:1) extracts. 5. Studied activity Insecticidal properties and insect growth regulatory effect against Spodoptera frugiperda by direct contact method [4], cytotoxicity by brine shrimp lethality test (BSLT) [5]. Criteria of evaluation. Larval mortality and LC50; larval weight, the growth index (GI), relative growth index (RGI), duration of pupal stage, and successfully pupation of S. frugiperda larvae [4] and LC50 of Artemia salina [5]. Data were analyzed by ANOVA [6], where results are given as probability values, with P b 0.05 adopted as the criterion of significance. Lethality was analyzed with a Finney computer program (Probit Analysis) to determine LC50. 6. Used organism S. frugiperda larvae used for the experiments were obtained from culture at the Centro de Investigación en Biotecnología at the Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico, maintained under previously described conditions [4].
Table 2 Effect of V. mollis leaf extracts on growth index (GI), relative growth index (RGI), duration of pupal stage, and successfully pupation of S. frugiperda and LC50 of A. salina Extracts
μg/ml
GI
RGI
Duration of pupal stage
Successful pupation %
BSLT a LC50
Control CH2Cl2
– 25 75 125 5 25 50 25 75 125
0.95 0.58 0.47 0.27 0.25 0.12 0.04 0.68 0.43 0.35
1.0 0.59 0.48 0.27 0.25 0.12 0.04 0.69 0.44 0.36
15.9 ± 0.16 13.2 ± 0.17 14.6 ± 0.25 13.8 ± 0.23 15.9 ± 0.28 13.6 ± 0.16 14.8 ± 0.40 13.9 ± 0.36 14.7 ± 0.41 12.3 ± 0.34
98 83 91 86 47 66 – 87 92 77
– 358.26
CHCl3–MeOH (1:1)
MeOH
a
BSLT, Brine Shrimp Lethality Test.
247.10
122.92
V. Rodríguez-López et al. / Fitoterapia 78 (2007) 37–39
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7. Results Reported in Tables 1 and 2. 8. Conclusion This study could contribute to the assessment of the possibility of using medicinal plants as potential insecticides [7]. The extracts were evaluated for their effect on the fall armyworm (S. frugiperda), an important insect pest of corn. All the extracts showed insecticidal and insect growth regulatory activity. The most active extract was CHCl3–MeOH (1:1) with LC50 13.63 ppm and 100% larval mortality at 75 ppm. On the other hand, this extract showed an important antifeeding activity against S. frugiperda (differences in larval weights were found between the control group and treatments group). Concerning insect growth inhibitory activity, the percentage of larvae that reached pupation decreased drastically with almost all extracts; larvae that reached pupal stage belonged to the lowest concentration group. The CHCl3– MeOH (1:1) extract had a 47% delay of pupation at 5 ppm, which was significant when compared with the data reported in the literature for methanolic extracts prepared from Yucca periculosa (25 ppm, 37.5%) used as the positive control [4]. Additionally, the extracts showed an important activity when tested to determine their toxicity to A. salina [5]. The results obtained have provided evidence to the potential use of V. mollis as insecticide. Bioassay-directed fractioning of the most active crude extracts to isolate and identify responsible compounds of the insecticidal activity and their possible mechanism of action is in progress. Acknowledgements The authors thank Biologist Laura Patricia Lina García and Biologist Mayra Pichardo for their valuable help in the biological assays. This study was financed by Grants from PROMEP-SEP and CONACyT (Convenio SEP-2003-CO243440/280). M. Z. Figueroa-Suárez acknowledges a scholarship (186641) from CONACyT. References [1] [2] [3] [4] [5]
Monroy O, Castillo E. Plantas medicinales utilizadas en el Estado de Morelos. Cuernavaca, México: CIB-UAEM; 2000. p. 260. Argueta A, Cano LM, Rodarte ME. Atlas de las plantas de la medicina tradicional, vol. I and III. México: Instituto Nacional Indigenista; 1994. Montiel-Herrera M, Camacho-Hernández IL, Ríos-Morgan A, Delgado-Vargas F. J Food Compos Anal 2004;17:205. Céspedes CL, Salazar JR, Martínez M, Aranda E. Phytochemistry 2005;66:2481. McLauglhlin JM. Crow gall tumors on potato disc and brine shrimp lethality: two simple bioassays for higher plant screening and fractionation. In: Hostettmann K, editor. Assays for bioactivity. San Diego: Academic Press; 1991. [6] Keppel G. Design and analysis: a researcher's handbook. New York: Prentice Hall; 1982. [7] Bowers SW. Insecticidal compounds from plants. In: Nigg HN, Seigler D, editors. Phytochemical resources for medicine and agriculture. New York: Plenum Publishing; 1992.