Natural essential oil compounds with heat treatment to control stem-end rot on grapefruit during postharvest handling and marketing

Project Overview

Project Type: Graduate Student
Funds awarded in 2015: $10,948.00
Projected End Date: 12/31/2016
Grant Recipient: University of Florida
Region: Southern
State: Florida
Graduate Student:
Major Professor:
Dr. Mark Ritenour
University of Florida

Annual Reports


  • Fruits: citrus


  • Pest Management: biorational pesticides, botanical pesticides, chemical control, physical control


    Screening of essential oils against mycelial growth of L. theobromae showed carvacrol and thymol had effective inhibition on mycelial growth of L. theobromae, with half –inhibition concentrations 0.045 and 0.037 mgŸmL-1, respectively. Both carvacrol and thymol in shellac coating exhibited effectively activities against diplodia stem end rot (SER) on ‘Ruby Red’ grapefruit when fruit were artificially inoculated with L. theobromae 12 h before coating application or immediately after coating application. Coating fruit with shellac containing carvacrol or thymol significantly decreased fruit natural decay, weight loss, and chilling injury during storage, but did not influence fruit internal quality.


    The grapefruit harvest season can stretch from late September to April or May. Early season grapefruit (September to December) usually reach internal maturity before the peel attains its characteristic external color. Therefore, a commercial degreening treatment that exposes fruit to 5 ppm ethylene at 28 to 29 °C with 90-95% relative humidity (RH) is often necessary because most consumers relate green color with immaturity (Ritenour et al., 2003a). However, the degreening environment can significantly increase diplodia stem-end rot (SER), caused by Lasiodiplodia theobromae, on citrus fruit (Brown, 1986; Zhang, 2004).

    Diplodia SER is often the most important postharvest disease of citrus fruit in warm and humid regions such as Florida.  Spores of L. theobromae infect immature citrus fruit on the tree and then remain quiescent in the necrotic floral tissue (Brown, 1971). After fruit maturity, L. theobromae can be isolated from most citrus fruit in Florida.  At or after harvest, hyphae grow through the developing abscission zone and into the peel, flesh, and central core of the fruit. Growth of L. theobromae is maximum at 30 °C, and RH at 95-100% can promote conidia germination (Barmore and Brown, 1985). During degreening, high temperature and RH conditions promote fungal growth on fruit, while ethylene speeds the formation of the abscission zone, reduces natural resistance of fruit tissue to pathogen, and promotes hyphal penetration into the flesh (Barmore and Brown, 1985; Brown and Burns, 1998). Without adequate control measures, diplodia SER can develop rapidly during degreening and subsequent transport to become serious problems upon arrival at destination markets.

    In Florida, thiabendazole (TBZ), imazalil (IMZ), and sodium-o-phenylphenate (SOPP) are commonly used to control postharvest decay on fresh citrus. However, all three have been used for more than three decades and their intense and repeated use can result in pathogen resistance to the fungicides, which has developed in many other regions of the world (Mari et al., 2014). In addition, consumer desires for reduced pesticide use and lower residues in general are encouraging investigations into more natural products (Zhang and Swingle, 2005). Furthermore, fungicide residues can limit citrus export because of lower maximum residue limits (MRLs) in many importing countries compared to the United States. For example, IMZ grapefruit MRLs are 10 ppm in the US, but only 5 ppm in Europe, Canada, and Japan. TBZ and SOPP grapefruit MRLs are 10 ppm in the US, but only 5 ppm in Europe. Therefore, effective and safe alternative methods to control postharvest decay of citrus are needed.

    In recent years, more attention has focused on exploitation of natural plant products that have little impact on human health or the environment to control postharvest disease. Essential oils are aromatic, oily liquids extracted from plant organs that are recognized as safe for human health and the environment (Antunes and Cavaco, 2010). Therefore, research into their use has expanded. Because of their antifungal activity, many essential oils have been reported to control postharvest disease caused by Penicillium spp., Alternaria spp., Botrytis cinerea, or Monilinia fructicola on a variety of fresh fruits and vegetables (Antunes and Cavaco, 2010). Jhalegar et al. (2015) found that dipping mandarin fruit in lemon grass oil, clove oil, eucalyptus, and neem oil significantly inhibited green (P. italicum) and blue (P. digitatum) mold. du Plooy et al. (2009) also reported that Mentha spicata and Lippia scaberrima essential oils, as well as pure limonene and carvone incorporated in wax coating showed significant effects against blue and green mold on oranges. Usually, the composition, structure, and functional group of essential oils play an important role in their antifungal activity, and compounds with phenolic groups are most effective (Dorman and Deans, 2000). Both carvacrol and thymol are phenolic compounds and have been reported to have antifungal activity against Botrytis cinerea, Monilinia fructicola, Rhizopus stolonifer, Alternaria arborescens, Geotrichum candidum, Colletotrichum acutatum, Botryodiplodia theobromae, Phytophthora cactorum, and Cryponectria parasitica (Kim et al, 2008; Numpaque et al., 2011; Plotto et al., 2002; Tsao and Zhou, 2000).

    Unfortunately, there can be drawbacks to applying essential oils on fresh fruit for postharvest decay control, such as the development of peel injury. Amiri et al. (2008) reported that dipping fruit in an aqueous 1 mgŸmL-1 eugenol solution resulted in peel injury on apple. Yan et al. (2016) also found that dipping fruit in an aqueous 2 mgŸmL-1 carvacrol or thymol solution resulted in phytotoxic peel injury on ‘Valencia’ oranges, but no injury developed when the same or even greater concentrations were incorporated into the wax coating. It was also reported that wax coating immediately after heat treatment greatly reduced the development of visible heat injury on ‘Marsh’ grapefruit. These coatings can slow fruit water loss and restrict oxygen diffusion to inhibit oxidative peel browning and symptom development (Ritenour et al., 2003b). Another advantage of incorporating essential oils into wax coatings is that essential oils are likely released slowly onto the product surface to better maintain their concentration during storage. In dipping treatment, the protective activity provided by essential oils decreases right after treatment because of their volatility. Kouassi et al. (2012) found that the effective concentration of Cinnamomum zeylanicum essential oil allowing 100% disease control was reduced to 0.5% after incorporating into shellac wax, while an effective concentration of C. zeylanicum essential oil into ethanol was 5% (Kouassi et al. 2010). In addition, coating citrus fruit with carnauba or shellac is an important process in Florida commercial packinghouse to reduce subsequent water loss and improve gloss, which also makes incorporating essential oils into wax practicable for commercial application.

    The objectives of this study were, 1) to evaluate the efficacy of carvacrol and thymol against Lasiodiplodia theobromae in vitro; and 2) to evaluate the ability of carvacrol and thymol, incorporated into a commercial shellac coating, to control diplodia SER on artificially inoculated ‘Ruby Red’ grapefruit.

    Literature Cited:

    Amiri, A., R. Dugas, A.L. Pichot and G. Bompeix. 2008. In vitro and in vitro activity of eugenol oil (Eugenia caryophylata) against four important postharvest apple pathogens. Int. J. Food Microsc. 126(1): 13-19.

    Antunes, M.D.C. and A.M. Cavaco. 2010. The use of essential oils for postharvest decay control. A review. Flavor Frag. J. 25: 351-366.

    Barmore, C.R. and G.E. Brown. 1985. Influence of ethylene on increased susceptibility of oranges to Diplodia natalensis. Plant Dis. 69: 228-230.

    Brown, G.E. 1971. Pycnidial release and survival of Diplodia natalensis spores. Phytopathology. 61:559-561.

    Brown, G.E. 1986. Diplodia stem-end rot, a decay of citrus fruit increased by ethylene degreening treatment and its control. Proc. Fla. State Hort. Soc. 99: 105-108.

    Brown, G.E. and J.K. Burns. 1998. Enhanced activity of abscission enzymes predisposes oranges to invasion by Diplodia natalensis during ethylene degreening. Postharvest Biol. Technol. 14: 217-227.

    Dorman, H.J.D. and S.G. Deans. 2000. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J. Appl. Microbiol. 88: 308-316.

    du Plooy, W., T. Regnier and S. Combrinck. 2009. Essential oil amended coatings as alternatives to synthetic fungicides in citrus postharvest management. Postharvest Biol. Technol. 53: 117-122.

    Jhalegar, M.J., R.R. Sharma and D. Singh. 2015. In vitro and in vivo activity of essential oils against major postharvest pathogens of Kinnow (Citrus nobilis× C. deliciosa) mandarin. J. Food Sci. Tech. 52(4): 2229-2237.

    Kim, J., Y.S. Lee, S.G. Lee, S.C. Shin and I.K. Park. 2008. Fumigant antifungal activity of plant essential oils and components from West Indian bay (Pimenta racemosa) and thyme (Thymus vulgaris) oils against two phytopathogenic fungi. Flavor Frag. J. 23(4): 272-277.

    Kouassi, K.H.S., M. Bajji and H. Jijakli. 2012. The control of postharvest blue and green molds of citrus in relation with essential oil-wax formulations, adherence and viscosity. Postharvest Biol. Technol. 73: 122-128.

    Kouassi, K.H.S., M. Bajji, A. Zhiri, P. Lepoivre and M.H. Jijakli. 2010. Evaluation of three essential oils as potential sources of botanical fungicides. Commun. Agric. Appl. Biol. Sci. 75(4): 525-529.

    Mari, M., A.D. Francesco and P. Bertolini. 2014. Control of fruit postharvest disease: old issue and innovative approaches. Stewart Postharvest Rev. 1:1-4.

    Numpaque, M.A., L.A. Oviedo, J.H. Gil, C.M. García and D.L. Durango. 2011. Thymol and carvacrol: biotransformation and antifungal activity against the plant pathogenic fungi Colletotrichum acutatum and Botryodiplodia theobromae. Trop. Plant Pathol. 36 (1):3-13.

    Plotto, A., D.D. Roberts and R.G. Roberts. 2002 Evaluation of plant essential oils as natural postharvest disease control of tomato (Lycopersicon esculentum). In XXVI International Horticultural Congress: Iss. Adv. in Postharvest Hort. 628: 737-745.

    Ritenour, A.M., M.M. William and W.W. Wardowski. 2003a. Recommendations for degreening Florida fresh citrus fruits. Florida Cooper. Ext. Serv. 1170: 1-3.

    Ritenour, M.A., K.J.J. Karuppiah, R.R. Pelosi, M.S. Burton, T.G. Mccollum, J.K. Brecht and E.A. Baldwin. 2003b. Response of Florida grapefruit to short-duration heat treatments using vapor heat or hot water dips. Proc. Fla. State Hort. Soc. 116: 405-409.

    Tsao, R. and T. Zhou. 2000. Antifungal activity of monoterpenoids against postharvest pathogens Botrytis cinerea and Monilinia fructicola. J. Essent. Oil Res. 12(1): 113-121.

    Zhang, J. 2004. Effect of ethylene on natural resistance of citrus fruit to stem-end rot caused by Diplodia natalensis and its relation to postharvest control of this decay. Proc. Fla. State Hort. Soc. 117: 364-367.

    Zhang, J. and P.P. Swingle. 2005. Effects of curing on green mold and stem-end rot of citrus fruit and its potential application under Florida packing system. Plant Dis. 89(8): 834-840.



    Project objectives:

    The over goal of this project is to evaluate the efficiency of natural compounds in ambient and heated solutions to control stem-end rot (SER), caused by L. theobromae, on grapefruit during postharvest handling and marketing.

    Objective 1. Screen essential oil compounds for their efficacy against L. theobromae in vitro. Study the effects of promising compounds to inhibit mycelium growth and conidial germination of L. theobromae.

    Objective 2. Using the most effective essential oil compounds from objective 1, evaluate their ability in both ambient and heated solutions to control SER on inoculated grapefruit and on naturally infected fruit.

    Objective 3. Test the effects of essential oil compounds in ambient or heated solutions on fruit internal and external quality during postharvest handling and simulated marketing conditions.

    Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture or SARE.