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Identification of Saturated Hydrocarbons from Jasmine (Jasminum sambac L.) Buds Damaged by Blossom Midge, Contarinia maculipennis Felt through GC-MS Analysis

I. Merlin Kamala , C. Chinniah, J.S. Kennedy, M. Kalyanasundaram, M. Suganthy
Applied Ecology and Environmental Sciences. 2017, 5(1), 10-18. DOI: 10.12691/aees-5-1-2
Published online: May 12, 2017

Abstract

Saturated hydrocarbons of fresh healthy jasmine buds (Jasminum sambac L) and infested buds damaged by blossom midge, Contarinia maculipennis Felt. were identified through Gas Chromatography-Mass spectrometry (GC-MS) to determine the saturated hydrocarbons. The results revealed that both the healthy and damaged buds had emitted hydrocarbon compounds numbering 24 and 34 respectively. The variation in the hydrocarbon constituents of healthy and damaged buds clearly depicts the emission of volatiles attracting beneficials witnessing tritrophic interactions in jasmine ecosystem. Linalool, the fragrant compound is detected in healthy jasmine buds at the retention time of 9.278 mins with the largest peak area of 5180722 mm2 and in midge damaged buds in a peak area of 288967 mm2, which might be the probable reason for the fragrance of jasmine flower. Allyl isothiocyanate, a compound responsible for plant defenses against herbivores is detected at a retention time of 4.306 mins recording the sixth largest peak area of 2461362 mm2 in midge infested buds. Other plant defense chemicals viz. naphthalene, azulene, methyl salicylate, methyl anthranilate, alpha farnescene, phenol, styrene etc. were also detected in midge damaged buds. The quality and quanity of these semiochemicals emitted by the buds might be the reason for attraction of natural enemies in the jasmine ecosystem there by further reducing the infestation of midge, as well as other pests. This feature can be exploited to enhance the efficacy of natural enemies in integrated management of jasmine pests.

1. Introduction

Jasmine is an important traditional flower, cultivated nearly throughout the tropical and subtropical parts of the world for its fragrant flowers. The flowers find place in floral decoration of marriages, to consecrate a sacred wedding ceremony, as a form of expressing love, affection, happiness and honouring the guests. The exchange of garlands made of Jasmine in marriage symbolizes the natural circle of protection and sacred bride to the spiritual life. Although more than 2,000 species are known, 40 been identified in India, and 20 species are cultivated in South India (Bhattacharjee, 1980). In India, jasmines are cultivated throughout the country. However, the largest area under Jasmine flower production is in Tamil Nadu followed by Karnataka. The annual production of flowers in India is worth more than 120 million (Dadlani, 2004). The world production of jasmine concrete is around 20 tonnes per annum, out of which India is producing and exporting about 2 tonnes (Ray et al., 2014). J.sambac is ravaged by several insects viz., budworm, blossom midge, leaf web worm, red spider mite and other sucking pests. Of these, budworm and blossom midge pose heavy damage as they directly attack the economic part. The adult midges lay eggs at the tip of the flower buds, when they are small and green in colour. The emerging maggots move down, reach the base of corollas and suck the sap, which results in swelling at the base of the buds and pink discolouration of buds. The infestation leads to premature drying of buds and leads to stunted growth and ultimately drying of plants. The management of jasmine pests completely relies on spraying of synthetic chemicals without right intervals, polluting the environment as well as causing detrimental effect to non- target beneficials. Therefore, it is now an urgent need to use safe but effective, biodegradable pesticides with no or less toxic effects on beneficials. An approach of using semiochemicals in pest management is to exploit ways to chemically augment, conserve or enhance the efficacy of natural enemies in a crop ecosystem.

Use of these biochemicals especially, the synomones released by host plants is of significance in biological control. Hydrocarbons present in host plants were found to act as synomones for natural enemies in different crop ecosystems. Plant surfaces are coated with thin layer of waxy materials that have myriad of functions. This layer is microcrystalline in structure and form the outer boundary of the cuticular membrane. It serves lot of purposes viz., to limit diffusion of water and solutes while permitting controlled release of volatiles that may deter the pests or attract natural enemies and pollinators (Eigenbrode, 1996). These plants waxy materials act as synomones and play a major role by guiding the natural enemies to the potential host or prey on the plant 9. Such clues may be utilized to stimulate foraging and host selection behavior of entomophages thereby increasing their effectiveness for IPM 1. Therefore in the present study, the hexane bud extracts of Jasmine damaged by midges and healthy buds were analyzed through GC-MS to determine the saturated hydrocarbon profiles in them.

2. Materials and Methods

Jasmine plants were raised in an area of 20 cents at the Botanical garden premises, Tamil Nadu Agricultural University, Coimbatore during January-May 2016. All the recommended agronomic practices were followed. Healthy and midge damaged buds were collected from the field during the period of heavy infestation by midges.

The saturated hydrocarbons were extracted from the buds infested by blossom midges and healthy buds using HPLC grade hexane as follows. The healthy and midge infested discoloured buds of jasmine were plucked carefully and used for extraction. Ten gram of buds was immersed overnight in 100 ml of HPLC grade hexane. The filtrate was then passed through silica gel (60-120 mesh) column. The hexane solvent was allowed to evaporate and the left over residue was collected by rinsing the container with a small quantity of HPLC grade hexane (Merck) and stored in separate vials for GC-MS Analysis.

Gas chromatography combined with mass spectroscopy is a preferable methodology for routine analysis of compounds. Hexane based bud extracts were analysed on GC-MS (Agilent Technologies 7890B GC System with 5977B MSD) mass selective detector (70eV) equipped with a 10:1 split injector. The gas chromatography is equipped with 30m fused silica capillary column having 0.25 mm ID and 0.25 um film thickness run in constant flow mode (1.0 ml/min helium). Oven temperature programming: 60°C (1 min hold) to 100°C at 5°C/min rate (1 min hold), then to 220°C at 10°C/min rate (5 min hold) and then to 240°C at 50°C/min rate (8 min hold). Injector temperature was set at 275°C. One microlitre of the extract was injected using auto sampler into the gas chromatography-mass spectroscopy (GC-MS) system for analysis and injections were done in split 10:1 mode. Agilent MSD productivity chemstation for GC- MS Systems data analysis applications software was used for the analysis of compounds in the extracts. Injected sample was separated into various constituents with different retention time which were detected by mass spectrophotometer. The compounds of interest were identified using standard NIST mass spectral (NIST MS 2) library. The chromatogram, a plot of intensity against retention time was recorded by the software attached to it. From the graph, the compounds were identified by comparing the data with the existing software libraries.

3. Results and Discussion

Induction of plant defense in response to herbivore involves the emission of volatile compounds called synamones that act as attractants for natural enemies of herbivores. Synamones produced by plants are reported to be very significant in eliciting host-seeking response in many natural enemies. Synamones attract predators and parasitoids, which elucidate the tritrophic interaction in a crop ecosystem. Gas Chromatography mass - spectroscopy analysis of synamone extracts of healthy buds showed the presence of 24 hydrocarbons, viz., cyclohexanol, cyclohexanone, 2-pentanol, 3- hexanol, 3- heptanol, 2-pentene, 5-nonanone, benzaldehyde, cyclopentanol, 3-hexenol acetate, benzyl alcohol, cyclohexane, heneicosane, linalool, phenyl methyl alcohol, methyl salicylate, alpha-farnescene, 9-tricosene, octadecane, tricosane, octacosane, tetracosane, eicosane and hentriacontane (Figure 1). Linalool was detected at the retention time of 9.278 mins with the largest peak area of 5180722 mm2 constituting 30.240 % of the total compounds. The hydrocarbons, octadecane, tricosane and octacosane was detected at the retention time of 27.552 mins with the second largest peak area of 4970489 mm2 constituting 7.618% of the total compounds. Hentriacontane, tetracosane and octacosane were detected at 3646033 mm2 recording the third largest peak area at RT 30.865 mins (Table 1).

But the midge damaged jasmine buds showed the presence of 34 saturated hydrocarbons viz., allyl isothiocyanate, styrene, 1,3 Pentadiene 4 methyl, benzoic acid, linalool, naphthalene, azulene, methyl salicylate, methyl anthranilate, tetradecane, alpha farnescene, naphthalene, phenol, hexadecane, heptadecane, heneicosane, octacosane, do triacontane 1-iodo, tetracosane, henetriacontane, pthalic acid isobutyloctyl ester, pentacosane, heptacosane, eicosane, indazole 4-one, 10-methyl antracene-9 carboxaldehyde, bis (2 ethyl hexyl phthalate), eicosane, tricosane, octacosane, tetracosane, hexadecane 1 - iodo, eicosane and nonacosane (Figure 2).

The results are in accordance with the findings of Kumar et al., 8, who reported that the hexane leaf extracts of targeted cole crop variety White Vienna subjected to gas liquid chromatography to detect their saturated hydrocarbon profile revealed the presence of three hydrocarbons viz., heneicosane (C21), docosane (C22) and nonacosane (C29).

The volatile compound linalool, is found to be present in the healthy and damaged buds of Jasmine (Figure 3). Linalool, a monoterpinol and the most common fragrance chemical is found to be present in Jasmine oil 2, 3, 4, 17. Linalool is detected in midge damaged buds in a peak area of 288797 mm2 at the retention time of 9.277 mins, as against 5180722 mm2 peak area in healthy jasmine buds implying its emission in large quantity in healthy buds. (Figure 3). This shows that linalool content is decreased in Jasmine due to midge attack. There are reports of more production of linalool due to herbivore attack. Linalool is the most abundant volatile emitted from fall army worm, Spodoptera frugiperda damaged rice plants 22.

The compound allyl iso-thiocyanate was detected at RT of 4.306 mins at an area of 2461362 mm2 in midge damaged buds, which is 5.070 per cent of the total compounds present. Allyl iso-thio cyanate, a naturally occurring organo-sulfur compound in mustard, radish, horseradish, is responsible for their pungent taste (Figure 4). Allyl isothiocyanate serves the plant as a defense against herbivores; since it is harmful to the plant itself, it is stored in the harmless form of the glucosinolate. When the plant is damaged, the enzyme myrosinase is released and acts on a glucosinolate known as sinigrin to give allyl isothiocyanate.

Synthetic allyl isothiocyanate is used as an insecticide, bacteriocide and nematocide, and is used in certain cases for crop protection 19. Zabża, 23; Titayavan and Altieri, 20, reported that the parasitoid Diaeretiella rapae M’Intoshis was attracted to allyl isothiocyanate emitted by cabbage plants damaged by cabbage aphids (Brevicoryne brassicae L.) and increased aphid parasitism from 8.5% to 22.5%. Thus, allyl isothiocyanate release due to midge attack in jasmine plant might attract lot of natural enemies that check the pest naturally due to tri trophic interactions. Allyl iso thiocyanate, had recorded sixth largest peak area and has already proved its efficiency in attracting natural enemies in various crop ecosystems.

Volatile methyl esters are common constituents of plant volatiles with important function in plant defense. Methyl salicylate, a herbivore-induced volatile has been shown to attract natural enemies and affect herbivore behavior. Methyl salicylate is detected in healthy jasmine buds itself at RT 12.002 mins and peak area of 263826 mm2 and in midge damaged buds in a peak area of 207138 mm2 showing the decrease in its content in midge damaged buds (Figure 5).

Methyl salicylate lures examined for its effectiveness against organic soybean aphids, Aphis glycines Matsumura showed reduced population of aphids, with significantly greater number of syrphid flies (Diptera: Syrphidae) and green lace wings (Neuroptera: Chrysopidae) 10. The results are in line with Du et al., 5; Kesseler and Baldwin, 7 reporting that a number of herbivory induced plant volatiles have been characterized for their individual contrition to indirect defense in behavioural set ups including methyl salicylate and linalool. Methyl anthranilate, a potential compound known to induce plant defense was detected in midge damaged buds at RT 14.849 mins in a peak area of 122329 mm2 (Figure 6).

Farnescene is produced by denovo biosynthesis in cotton plants when damaged by insect herbivores 11. These compounds are likely to mediate the interactions between herbivores and their natural enemies, attracted by terpenes.

Alpha farnescene is detected in healthy jasmine buds at RT 17.305 mins and peak area of 215542 mm2 constituting 0.330 percent and in midge infested buds in a peak area of 372035 mm2 at the retention time 17.039 mins constituting 0.391 percent (Figure 7).

Napthalene, an aromatic hydrocarbon was detected twice in midge infested buds at RT 11.657 mins with peak area of 633398 mm2, and at RT 17.493 mins with peak area of 222876 mm2 which was reported to be a semiochemical attracting natural enemies of stemborer in maize ecosystem 13 (Figure 8).

The hydrocarbons, tricosane, octacosane and tetracosane (Figure 9, Figure 10, Figure 11) recorded the largest peak area of 14863097 mm2 at 27.554 mins of retention time and constituting 22.777% of the total compounds which implies their emission in maximum quantity. The hydrocarbon, hexadecane 1 - iodo recorded a peak area of 7789615 mm2 at RT 29.047 mins. The hydrocarbon eicosane, was released at RT 30.868 mins with the second largest peak area of 5018550 mm2 constituting 7.728 of total area and again at 329550 mm2 at RT 27.251 mins. The hydrocarbon, heptacosane recorded a peak area of 4547459 mm2 at RT 22.0153 mins showing 5.847 % of total emission. Phenol is emitted at RT 17.362 mins occupying a peak area of 3059318 mm2.

The compound, pthalic acid isobutyloctyl ester was detected at a peak area of 2880188 mm2 at RT 21.531 mins, heneicosane and octacosane at 2372226 mm2 at 19.773 mins octadecane at 20.730 mins in a peak area of 2140636 mm2 and pentacosane at 2119427 mm2 in RT 21.774 mins.

The GC-MS studies on the volatile profile emitted from leaf folder damaged leaves in the susceptible rice variety TN1 showed more of the presence of docosane, which could be responsible in attraction of Trichomma cnaphalocroccis Uchida and Cotesia angustibasis Gahan 15.

Seenivasagan and Paul 18 analyzed the extracts of cruciferous host plants of diamond back moth and revealed the presence of saturated hydrocarbons. Cauliflower leaf extract contain 12 hydrocarbons with carbon number ranging from C10-C30 in which C29 (nonacosane) was detected in highest quantity. In cauliflower extracts exclusively C10 (decane) and C12 (dodecane) hydrocarbons were identified which were not detected in other host plant extracts. The hydrocarbon C14 (tetradecane) was detected only in cauliflower and broccoli extracts, whereas C16 hexadecane was detected only in cabbage, cauliflower and broccoli extracts. C18 (octadecane) and C20 (eisocane) were detected in cabbage and cauliflower extracts. C22 (docosane) and C25 (pentacosane) were detected only in cauliflower, while C26 (hexacane) was found only on knoll-knol leaf extracts. Similarly hexane extracts of ten different varieties of tomato (Lycopersicon esculentum Mill.) obtained in the vegetative and flowering phase of growth contained tricosane (C23), heneicosane (C 21), pentacosane (C25) and hexacosane (C26) during the vegetative period and heneicosane (C21) and hexacosane (C26) during the flowering period 12.

Comparing the peak areas of hydrocarbon compounds present in healthy and damaged buds, six compounds have more peak area in damaged than healthy buds. Alpha farnescene has recorded 215542 mm2 peak area at RT 17.305 mins in healthy buds, but has recorded 372035 mm2 in midge damaged buds recording 1.72 per cent increase in peak area. In healthy buds, the saturated hydrocarbons, tricosane and octacosane has recorded a peak area of 4970489 mm2 at 27.552 mins, as against 14863097 mm2 in midge damaged buds, recording 2.97 per cent increase.

Also, saturated hydrocarbon, tetracosane recorded 1432970 mm2 peak area in healthy buds at RT 29.056 mins as against 14863097 mm2 peak area in midge damaged buds recording 10.07 per cent increase. The hydrocarbon, eicosane was detected at 29.046 mins with a peak area of 1432970 mm2 in healthy buds and was detected thrice in midge damaged buds at RT 22.830 mins, 27.251 mins and 30.868 mins in a peak area of 1483204, 3295502 and 5018550 mm2 respectively with 1.4 and 1.5 per cent increase in release. The content of the plant defense compound, alpha farnescene present in midge damaged buds was higher than the healthy ones. Thus to summarize, midge damaged buds have more peak area when compared to the healthy jasmine buds, witnessing the increased production of hydrocarbons, which in turn attracts natural enemies.

Behaviour of a natural enemy can be manipulated potentially by enhancing their foraging ability in an ecosystem. The interface, where tritrophic interactions take place in natural condition is often the cuticle of a plant. The saturated hydrocarbons present in the epicuticular wax layer of plants have been shown to influence the foraging success of natural enemies. Therefore the hydrocarbon compounds found in the extracts of midge damaged buds have to be explored for the attraction of natural enemies to parasitize and/or to predate the herbivore or reduce the further infestation by the herbivores for efficient pest management.

Acknowledgements

The financial assistance provided by UGC, fellowship to pursue Ph.D. in Agricultural Entomology at Tamil Nadu Agricultural University, Coimbatore is gratefully acknowledged for the senior author.

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In article      View Article
 
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Cite this article:

Normal Style
I. Merlin Kamala, C. Chinniah, J.S. Kennedy, M. Kalyanasundaram, M. Suganthy. Identification of Saturated Hydrocarbons from Jasmine (Jasminum sambac L.) Buds Damaged by Blossom Midge, Contarinia maculipennis Felt through GC-MS Analysis. Applied Ecology and Environmental Sciences. Vol. 5, No. 1, 2017, pp 10-18. http://pubs.sciepub.com/aees/5/1/2
MLA Style
Kamala, I. Merlin, et al. "Identification of Saturated Hydrocarbons from Jasmine (Jasminum sambac L.) Buds Damaged by Blossom Midge, Contarinia maculipennis Felt through GC-MS Analysis." Applied Ecology and Environmental Sciences 5.1 (2017): 10-18.
APA Style
Kamala, I. M. , Chinniah, C. , Kennedy, J. , Kalyanasundaram, M. , & Suganthy, M. (2017). Identification of Saturated Hydrocarbons from Jasmine (Jasminum sambac L.) Buds Damaged by Blossom Midge, Contarinia maculipennis Felt through GC-MS Analysis. Applied Ecology and Environmental Sciences, 5(1), 10-18.
Chicago Style
Kamala, I. Merlin, C. Chinniah, J.S. Kennedy, M. Kalyanasundaram, and M. Suganthy. "Identification of Saturated Hydrocarbons from Jasmine (Jasminum sambac L.) Buds Damaged by Blossom Midge, Contarinia maculipennis Felt through GC-MS Analysis." Applied Ecology and Environmental Sciences 5, no. 1 (2017): 10-18.
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  • Table 1. Saturated hydrocarbon profile of the healthy and blossom midge damaged buds of Jasmine, J.sambac
[1]  Ahmad, F., M. Aslam and M. Razaq. 2004. Chemical ecology of insects and tritrophic interactions. Journal of Research Science., 15: 181-190.
In article      View Article
 
[2]  Cockayne, S.E., and D.J. Gawkrodger. 1997. Occupational contact dermatitis in an aromatherapist. Contact Dermatitis., 37: 306-307.
In article      View Article
 
[3]  De Groot. A.C., and S. Frosch. 1987. Contact allergy to cosmetics: causative ingredients. Cont. Derm., 17: 26-34. 63.
In article      View Article
 
[4]  De Groot, A.C., P.J. Coenraads, D.P. Bruynzeel, B.A. Jagtman, C.J.W. van-Ginkel, K. Noz, P.G.M. Van Der S. Pavel, J. Vink and J.W. Weyland. 2000. Routine patch testing with fragrance chemicals in The Netherlands. Cont. Derm., 42: 184-185.
In article      View Article
 
[5]  Du, Y.J., G. M. Poppy, W. Powell, J. A. Pickett. L. J. Wadhams, C. M. Woodcocks. 1998. Identification of semiochemicals released during aphid feeding that attract parasitoid, Aphidius ervi., J. Ecol., 24: 1355-1365.
In article      View Article
 
[6]  Gunasekaran, V. 1989. Studies on bio-ecology of jasmine pest complex. M. Sc (Ag.) Thesis, Tamil Nadu Agric. University, Coimbatore.
In article      
 
[7]  Kesseler, A., and I.T. Baldwin. Defensive function of herbivore-induced plant volatile emissions in nature. Science., 291: 2141-2144.
In article      View Article  PubMed
 
[8]  Kumar, A., A. Zayeem and S. Kanameni.2012. Synamonal effect of cole crops of individual and associative learning behavior of Cotesia plutellae. International Journal of biology., Pharmacy and AlliedSciences (IJBPAS) 1: 285-298.
In article      View Article
 
[9]  Hilker, M., and T.Meiners.2006. Early herbivore alert: insect eggs induce plant defense. Journal of chemical Ecology., 32: 1379-1397.
In article      View Article  PubMed
 
[10]  Malinger, R.E., D.B. Hogg and C. Horton.2011. Methyl salicylate attracts natural enemies and reduces populations of soybean aphids (Hemiptera: Aphididae) in soybean agroecosystems. J. Eco. Ento., 104(1): 115-124.
In article      View Article
 
[11]  Pare, P.W., and J.H.Tumilson.1997. Denova biosynthesis of volatiles induced by insect herbivory in cotton plants. Plant. Physiol., 114: 1161-1167.
In article      View Article  PubMed
 
[12]  Paul, A.V.N. Srivastava, M., P.Dureja and A.K.Singh. 2008. Semiochemicals produced by tomato varieties and their role in parasitism of Corcyra cephalonica (Lepidoptera: pyralidae) by the egg parasitoid Trichogramma chilonis (Hymenoptera: Trichogrammatidae). International Journal of Tropical Insect Science., 28: 10-116.
In article      View Article
 
[13]  Peshwin, R., and D. Pimental. 2014. Integrated pest Management: Experiences with Implementation. Vol 4. Springer publication.
In article      
 
[14]  Prakash, K and B. Muniyandi. 2014. Application of ARIMA Model for Forecasting Production of Jasmine Flower in Madurai District of Tamil Nadu, India. American International Journal of Research in Humanities, Arts and Social Sciences., 6(3): 279-285.
In article      
 
[15]  Rathika, M. and R. Nalini. 2011. Role of rice plant volatiles on the orientation of leaf folder larval parasitoids Trichomma cnaphalocroccis Uchida and Cotesia angustibasis Gahan. Oryza-An International Journal of Rice., 48(3): 250-254.
In article      View Article
 
[16]  Romanowski, F., and H. Klenk.2005. “Thiocyanates and Isothiocyanates, Organic”, Ullmann's Encyclopedia of Industrial Chemistry, Weinheim: Wiley-VCH.
In article      
 
[17]  Schaller, M., and H.C. Korting. 1995. Allergic airborne contact dermatitis from essential oils used in aroma therapy. Clin. Exp. Dermatol., 20:143-145.
In article      View Article  PubMed
 
[18]  Seenivasagan, T. and A.V.N. Paul.2011. Gas chromatography and electroantennogram analysis of saturated hydrocarbons of cruciferous host plants and host larval body extracts of Plutella xylostella for behavioural manipulation of Cotesia plutellae. Indian Journal of Experimental biology., 49: 375-386.
In article      PubMed
 
[19]  Shoram, N.S. Parekh, N.V. Upadhyay, B. A. Karapatiya and H. C. Patel. 2012. Effect of nitrogen and phosphorus on vegetative growth and flower yield of Jasmine. The Asian Journal of Horticulture., 7(1):52-54.
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