Azadirachta indica and Citrullus colocynthis extracts increase defense response of wounded Ricinus communis and improve its growth

Suzan A. Sayed, Mohamed A. A. Gadallah


The interactive effects of mechanical wounding with or without crude extracts of neem (Azadirachta indica) and bitter apple (Citrullus colocynthis,  CCT) supplementation on growth, chlorophyll, carotenoids, soluble sugars (SS), soluble proteins (SP) and total free amino acids (TAA) in Ricinus communis plants were studied. In responses to mechanical wounding Ricinus plants produced more soluble sugars and soluble proteins. On the other hand, chlorophyll a, cartenoids and total free amino acids contents as well as dry mass production declined upon wounding. Neem and CCT crude extracts application, whether independently or in combination, counteracted in various degrees the deleterious effects of wounding stress on growth. Crude extracts increased SS, SP as well as Chl and cartenoids contents and improved wounded plants growth. The effect of single factors (wounding, neem and CCT extract) could be modified or reverse by the interaction between these factors when used in combination (eg. total amino acids, Chl a and carotenoids). The results clearly indicate that CCT and neem crude extracts supplementation might be beneficial in attenuating the harmful effects of mechanical wounding stress on plant growth.


Amino acids; Carotenoids; Chlorophyll; Leaf area; Soluble sugars

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Cheong YH, Chang HS, Gupta R, Wang X, Zhu T, Luan S. Transcriptional profiling reveals novel interactions between wounding, pathogen, abiotic stress, and hormonal responses in Arabidopsis. Plant Physiol. 2002; 129: 661-677.

Łukaszuk E, Ciereszko I. Plant responses to wounding stress. In: Łaska G, ed. Biological diversity - from cell to ecosystem. Polish Botanical Society - Branch in Białystok. 2012: 73-85.

Sayed SA, Gadallah MAA. Effects of crude plant extracts on wounded Ricinus communis plants. Eur J Biol Res. 2016; 6(2): 82-91.

Choi Y, Tomas-Barberan FA, Saltveit ME. Wound-induced phenolic accumulation and browning in lettuce (Lactuca sativa L.) leaf tissue is reduced by exposure to n-alcohols. Postharvest Biol Technol. 2005; 37: 47-55.

Jacobo-Velázquez DA, Cisneros-Zevallos L. An alternative use of horticultural crops: stressed plants as bio-factories of bioactive phenolic compounds. Agriculture. 2012; 2: 259-271.

Bari R, Jones JD. Role of plant hormones in plant defence responses. Plant Mol Biol. 2009; 69: 473-488.

Santner A, Estelle M. Recent advances and emerging trends in plant hormone signalling. Nature. 2009; 459: 1071-0180.

Smith JL, De Moraes CM, Mescher MC. Jasmonate- and salicylate-mediated plant defense responses to insect herbivores, pathogens and parasitic plants. Pest Manag Sci. 2009; 65: 497-503.

Demkura PV, Abdala G, Baldwin IT, Ballare CL. Jasmonate-dependent and independent pathways mediate specific effects of solar ultraviolet B radiation on leaf phenolics and antiherbivore defense. Plant Physiol. 2010; 152: 1084-1095.

Larrieu A, Vernoux T. How does jasmonate signaling enable plants to adapt and survive? BMC Biology. 2016; 14: 79.

Lomate PR, Hivrale, VK. Induction of leucine aminopeptidase (LAP) like activity with wounding and methyl jasmonate in pigeonpea (Cajanas cajan) suggests the role of these enzymes in plant defense in Leguminosae. Plant Physiol Biochem.n2011; 49: 609-616.

Giridhar KV, Thimann G. The interaction of senescence and wounding in oat leaves. II. Chlorophyll breakdown caused by wounding in light. Plant Sci. 1988; 54: 133-139.

Junior FM, Fernandes IM, Santos CS, Xavier de Mesquita L, Pereira RA, Maracaja PB, Soto-Blanco B. Toxicity of castor bean (Ricinus communis) pollen to honeybees. Agric Ecosyst Environ. 2011; 141: 221-223.

Singh RK, Gupta MK, Singh AK, Kumar S. Pharmacognostical investigation of Ricinus communis stem. IJPSR. 2010; 1(6): 89-94.

Rana M, Dhamija H, Prashar B, Sharma S. Ricinus communis L. A review. Int J Pharm Tech Res. 2012; 4(4): 1706-1711.

Karim A, Nouman M, Munir S, Sattar S. Pharmacology and phytochemistry of Pakistani herbs and herbal drugs used for treatment of diabetes. Int J Pharmacol. 2011; 7: 419-439.

Al-Snafi AE. Chemical constituents and pharmacological effects of Citrullus colocynthis. A review. IOSR-PHR. 2016; 6(7): 17-31.

Schmutterer H. The neem tree. Publisher VCH, Germany and VCH Publishers Inc. New York, USA. 1995.

Hasmat AI, Azad H, Ahmed A. Neem (Azadirachta indica A. Juss) - a nature’s drugstore: an overview. I Res J Biol Sci. 2012; 1: 76-79.

Hossain MA, Al-Toubi WAS, Weli AM, Al-Riyami QA, Al-Sabahi JN. Identification and characterization of chemical compounds in different crude extracts from leaves of Omani neem. J Taibah Univ Sci. 2013; 7: 181-188.

Pandey G, Verma KK, Singh M. Evaluation of phytochemical, antibacterial and free radical scavenging properties of Azadirachta indica (neem) leaves. Int J Pharm Pharm Sci. 2014; 6: 444-447.

Del Serrone P, Toniolo C, Nicoletti M. Neem (Azadirachta indica A. Juss) oil: a natural control meat spoilage. Foods. 2015; 4: 3-14.

Gopal M, Gupta A, Arunachalam V, Magu SP. Impact of azadirachtin, an insecticidal allelochemical from neem on soil microflora, enzyme and respiratory activities. Biores Technol.2007; 98: 3154- 3158.

Zhang Y, Xu J, Yin Z, Jia R, Lu Y, Yang F, et al. Isolation and identification of the antibacterial active compound from petroleum ether of neem oil. Fitoterapia. 2010; 81: 747-750.

Sukanya SL, Sudisha J, Hariprasad P, Niranjana SR, Prakash HS, Fathima SK. Antimicrobial activity of leaf extracts of Indian medicinal plants against clinical and phytopathogenic bacteria. Afr J Biotechnol. 2009; 8: 6677-6682.

Al-Hazmi RHM. Effect of neem (Azadirachta indica) leaves and seeds extract on the growth of six of the plant disease causing fungi. Glob Adv Res J Microbiol. 2013; 2: 89-98.

Jain D, Jayaram L, Prabhu MV, Bhat KG. Antibacterial effect of neem (Azadirachta indica) oil on multidrug resistant bacteria isolated from human infections. Int J Biol Med Res. 2013; 4: 3544-3546.

Beck JJ, Smith L, Merrill GB. In situ volatile collection, analysis, and comparison of three Centaurea species and their relationship to biocontrol with herbivorous insects. J Agr Food Chem. 2008; 56: 2759-2764.

Yu JQ, Ye SF, Zhang MF, Hu WH. Effects of root exudates and aqueous root extract cucumber (Cucumis sativus) and allelochemicals, on photosynthesis and antioxidant enzymes in cucumber. Biochem Syst Ecol. 2003; 31: 129-139.

Lichtenthaler K. Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. In: Packer L, Douce R, eds. Methods in Enzymology. Academic Press, New York. 1987; 148: 350-382.

Bates LS, Waldren RP, Teare ID. Rapid determination of free proline for water stress studies. Plant Soil. 1973; 39: 205-207.

Buysse J, Merckx R. An improved colorimetric method to quantify sugar content of plant tissue. J Exp Bot. 1993; 44: 1627-1629.

Lee YP, Takahashi T. An improved colorimetric determination of amino acids with the use of ninhydrin. Anal Biochem. 1966; 14: 71-77.

Lowry OH, Resbrough NJ, Farr AL, Randall RJ. Protein measurement with the folin-phenol reagent. J Biol Chem. 1951; 193: 265-275.

Watason DI, Watson MA. Studies in potatoes agronomy. Effect of variety, seed size and spacing on growth, development and yield. J Agr Sci. 1953; 66: 241.

Duncan DB. Multiple ranges and multiple F-test. Biometrics. 1955; 11: 1-42.

Ostle B. Statistics in research. Iowa State University Press, Ames. 1963.

Rakwal R, Agrawal GK. Wound signaling-coordination of the octadecanoid and MAPK pathways. J Physiol Biochem. 2003; 41: 855-861.

Becerra-Moreno A, Redondo-Gil M, Benavides J, Nair V, Cisneros-Zevallos L, Daniel A, Jacobo-Velázquez DA. Combined effect of water loss and wounding stress on gene activation of metabolic pathways associated with phenolic biosynthesis in carrot. Front Plant Sci. 2015; 6(837): 1-15.

Zhang Y, Turner JG. Wound-induced endogenous jasmonates stunt plant growth by inhibiting mitosis. PLoS ONE. 2008; 3(11): e3699.

Li ZH, Wang Q, Ruan X, Pan CD, Jiang DA. Phenolics and plant allelopathy. Molecules. 2010; 15: 8933-8952.

Lotina-Hennsen B, King-Diaz B, Aguilar MI, Terrones MGH. Plant secondary metabolites. Targets and mechanisms of allelopathy. In: Reigosa MJ, Pedrol N, Gonzalez L, eds. Allelopathy: a physiological process with ecological implications. The Netherlands, Springer. 2006: 229-265.

Chou CH. Introduction to allelopathy. In: Reigosa MJ, Pedrol N, Gonzalez L, eds. Allelopathy: a physiological process with ecological implications. The Netherlands, Springer. 2006: 1-9.

Salim AH, Jasim NB. Cost benefit ratio of infected tomato yield by Fusarium wilt disease. IJRANSS. 2016; 4: 103-108.

Salama HMH, Al Rabiah HKA. Physiological effects of allelopathic activity of Citrullus colocynthis on Vicia faba and Hordeum vulgare. Eur J Biol Res. 2015; 5(2): 25-35.

Umarani R, Vanangamudi K. Pre-storage treatments to improve viability in Casuarina equisetifolia seeds. Madras Agric J. 2005; 92: 7-9.

An M, Johnson IR, Lovett IR. Mathematical modeling of allelopathy: biological response to allelochemicals and its interpretation. J Chem Ecol. 1993; 19: 2379-2388.

Gajalakshmi S, Abbasi SA. Neem leaves as a source of fertilizer-cum-pesticide vermicompost. Biores Technol. 2004; 92: 291-296.

Mamman PH, Mshella WP, Susbatrus SC, Sambo KW. Antibacterial effects of crude extract of Azadirachta indica against Escherichia coli, Salmonella spp. and Staphylococcus aureus. Int J Med Med Sci. 2013; 5: 14-18.

Nwankwo EN, Onuseleogu DC, Ogbonna Confidence U, Okorocha AOE. Effect of neem leaf extracts (Azadirachta indica) and synthetic pesticide (carbofuran) on the root-knot nematode (Meloidogyne spp.) of cowpea (Vigna unguiculata L. Walp). Int J Entomol Res. 2016; 1(3): 1-6.

Lukaszuk E, Rys M, Mozdzen K, Stawoska I, Skoczowski A, Ciereszko I. Photosynthesis and sucrose metabolism in leaves of Arabidopsis thaliana aos, ein4 and rcd1 mutants as affected by wounding Acta Physiol Plant. 2017; 39: 17.

García-Plazaola JI, Portillo-Estrada M, Fernández-Marín B, Kännaste A, Niinemets Ü. Emissions of carotenoid cleavage products upon heat shock and mechanical wounding from a foliose lichen. Environ Exp Bot. 2017; 133: 87-97.

Gill SS, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem. 2010; 48: 909-930.

Havaux M. Carotenoid oxidation products as stress signals in plants. Plant J. 2014; 79(4): 597-606.

Othman R, Mohd Zaifuddin FA, Hassan NM. Carotenoid biosynthesis regulatory mechanisms in plants. J Oleo Sci. 2014; 63(8): 753-760.

Hesse H, Willmitzer L. Expression analysis of a sucrose synthase gene from sugar beet (Beta vulgaris L.). Plant Mol Biol. 1996; 30: 863-872.

Klotz KI, Haagenson MD. Wounding, anoxia and cold induce sugar beet sucrose synthase transcriptional changes that are unrelated to protein expression and activity. J Plant Physiol. 2008; 165: 423-434.

Boege K. Influence of plant ontogeny on compensation to leaf damage. Am J Bot. 2005; 92: 1632-1640.

El-Khatib AA, Hegazy AK. Growth and physiological responses of wild Oats allelopathic potential of wheat. Acta Agronom Hungar. 1999; 47(1): 11-18.

El-Khawas SA, Shehata MM. The allelopathic potentialities of Acacia nilotica and Eucalyptus rostrata on monocot (Zea mays L.) and dicot (Phaseolus vulgaris L.) plants. Biotechnol. 2005; 4(1): 23-34.

Berger S, Sinha AK, Roitsch T. Plant physiology meetsphytopathology: plant primary metabolism and plant-pathogen interactions. J Exp Bot. 2007; 58: 4019-4040.

Rai VK. Role of amino acids in plant responses to stresses. Biol Plantarum. 2002; 45: 481-487.

Roychoudhur A, Banerjee A, Lahiri V. Metabolic and molecular-genetic regulation of proline signaling and its cross-talk with major effectors mediates abiotic stress tolerance in plants. Turk J Bot. 2015; 39: 887-910.

Yaish MW. Proline accumulation is a general response to abiotic stress in the date palm tree (Phoenix dactylifera L.). Genet Mol Res. 2015; 14(3): 9943-9950.

Liang X, Zhang L, Natarajan SK, Becker DF. Proline mechanisms of stress survival. Antioxid Redox Signal. 2013; 19(9): 998-1011.

Natarajan SK, Zhu W, Liang X, Zhang L, Demers AJ, Zimmerman MC, et al. Proline dehydrogenase is essential for proline protection against hydrogen peroxide-induced cell death. Free Radic Biol Med. 2012; 53: 1181-1191.

Liu W, Hancock CN, Fischer JW, Harman M, Phang JM. Proline biosynthesis augments tumor cell growth and aerobic glycolysis: involvement of pyridine nucleotides. Sci Rep. 2015; 5: 17206.

Phang JM, Liu W, Hancock C, Christian KJ. The proline regulatory axis and cancer. Front Oncol. 2012; 2: 60.


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