C O N T E N T S JOURNAL OF HORTICULTURAL SCIENCES Volume 16 Issue 1 June 2021 In this Issue i-ii Review Moringa (Moringa oleifera L.): An underutilized and traditionally valued 1-13 tree holding remarkable potential Jattan M., Kumari N., Raj Kumar, Kumar A., Rani B., Phogat D.S., Kumar, S. and Kumar, P. Original Research in Papers Characterization and evaluation of mountain sweet thorn 14-25 (Flacourtia montana J. Grah) collections Tripathi P.C., Ganeshan S., Radhika V. and Shetti D.L. Optimization of methodology for the extraction of polyphenolic compounds 26-35 with antioxidant potential and á-glucosidase inhibitory activity from jamun (Syzygium cumini L.) seeds Arivalagan M., Priyanka D.R. and Rekha A. Genetic variability studies in amaranthus (Amaranthus spp.) 36-44 Agadi A.H., Kolakar S., Lakshmana D., Nadukeri S. and Hanumanthappa M. Morpho-physiological parameters associated with chlorosis resistance to 45-52 iron deficiency and their effect on yield and related attributes in potato (Solanum tuberosum L.) Challam C., Dutt S., Sharma J., Raveendran M. and Sudhakar D. Responses of different Okra (Abelmoschus esculentus) cultivars to water 53-63 deficit conditions Ayub Q., Khan S.M., Hussain I., Naveed K., Ali S., Mehmood A., Khan M.J., Haq N.U., Shehzad Q. Induced variability for yield and its attributing traits in cluster bean 64-68 [Cyamopsis tetragonoloba (L. ) Taub] through gamma irradiation Lavanya H.N., Mishra S., Sood M., Aghora T.S., Anjanappa M., Rao V.K. and Reddy A.B. In vitro multiplication protocol for Curcuma mangga : Studies on carbon, 69-76 cytokinin source and explant size Waman A.A., Bohra P., Karthika Devi R. and Pixy J. Effect of fungicide and essential oils amended wax coating on quality and shelf life 77-90 of sweet orange (Citrus sinensis Osbeck) Bhandari M., Bhandari N. and Dhital M. Post-harvest quality and quantification of betalains, phenolic compounds and 91-102 antioxidant activity in fruits of three cultivars of prickly pear (Opuntia ficus-indica L. Mill) Gonzalez F.P.H., Saucedo V.C., Guerra R.D., Suarez E.J., Soto H.R.M. Lopez J.A., Garcia C.E. and Hernandez R.G. Soil microbial community dynamics as influenced by integrated nutrient 103-113 management practices in sweet basil (Ocimum basilicum L.) cultivation Baraa AL-Mansour and D. Kalaivanan Effect of spectral manipulation and seasonal variations on cut foliage production 114-120 and quality of Philodendron (Philodendron ‘Xanadu’) Sujatha A. Nair, Laxman R.H. and Sangama Short Communications Studies on mutagenic sensitivity of seeds of pummelo (Citrus maxima Merr.) 121-124 Sankaran M., Kalaivanan D. and Sunil Gowda D.C. Isolation and characterization of microsatellite markers from 125-129 Garcinia indica and cross species amplification Ravishankar K.V., Vasudeva R., Hemanth B., Nischita P., Sthapit B.R., Parthasarathy V.A. and Rao V.R. 77 J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Original Research Paper Effect of fungicide and essential oils amended wax coating on quality and shelf life of sweet orange (Citrus sinensis Osbeck) Bhandari M.1, Bhandari N.*1and Dhital M.2 1Institute of Agriculture and Animal Science, Rampur, Chitwan, Nepal 1*Institute of Agriculture and Animal Science, Gauradaha, Jhapa, Nepal 2Agriculture and Forestry University, Rampur, Chitwan, Nepal *Corresponding author e-mail: iaasnirajan@gmail.com ABSTRACT Laboratory research was conducted to study the effect of wax amended coating on the shelf life of Citrus sinensis Osbeck during 2017-18 at Rampur, Chitwan. The experiment was conducted in single factor Completely Randomized Design (CRD) with nine treatments and four replications. The treatments consisted of carbendazim and three essential oils viz. lemongrass, mentha and eucalyptus oil at two different concentrations of 0.1% and 0.5%, all of them infused with 10% wax emulsion. The wax treatment devoid of fungicide and essential oils served as control. The application of essential oils with wax improved shelf life and enhanced juice retention, firmness, titratable acidity, vitamin C and disease reduction. But total soluble solid was found higher in fruits treated with wax emulsion only. The highest shelf life and disease control was obtained with wax with 0.5% carbendazim but waxing with 0.5% eucalyptus oil and 0.5% lemongrass oil can be better alternatives considering their superior performance in environmental aspects, consumer preferences and quality parameters like juice retention, firmness, titratable acidity and vitamin C. Keywords : Carbendazim, Eucalyptus oil, Green mold, Lemongrass oil, Post-harvest INTRODUCTION Sweet Or a nge (Citrus sinensis Obseck) is a n economically important citrus fruit of the mid hill region of Nepal. The mid hill region of Nepal (1000 to 1500 masl altitude) has a comparative advantage in the production of sweet orange over traditional crops (rice, wheat, maize etc). Sweet orange is the second most gr own citrus crop in Nepa l after Mandarin in terms of area and production (MOALD, 2020). The oranges in the Nepalese agricultural market have to compete with products coming from neighboring countries like India and China. The cost of production of sweet orange is higher due to high input costs, the need for hybrid budded and grafted saplings and intensive labor requirements to grow the crop which has forced the grower to think about improving postharvest management practices. The lack of suitable storage and preservation techniques forces the farmers to sell sweet oranges before their horticultural maturity and just after picking. The unaffordable postharvest preservation has led to a negative effect on the citrus enterprises in Nepal (Kaini, 2013). Green mold (Penicillium digitatum Sacc.) and blue mold (Penicillium italicum Wehmer) are the most economically important postharvest pathogens of sweet orange causing significant losses (Abd-El-Khair and Hafez, 2006; El-Otmani et al., 2011; Papoutsis et al., 2019). Currently, the control of green and blue mold is a ccomplished by pr e-a nd postha r vest a pplica tion of chemica l fungicides such a s carbendazim, imazalil, thiabendazole, pyrimethanil, fludioxonil, prochloraz and and guazatine (Danderson, 1986; Ismail and Zhang, 2004; Smilanick et al., 2006; Smilanick et al., 2008; Berk, 2016; Joshi et al., 2020). Br oadly, such fungicides inhibit the ergoster ol synthesis, mitochondrial electron tra nsport and synthesis of multi-site enzymes, protein and nucleic acid thereby kill or inhibit fungi or fungal spore This is an open access article d istributed under the terms of Creative Commons Attribution-NonCommer cial-ShareAl ike 4.0 International License, which permits unrestricted non-commercial use, d istribution, and reproduction in any med ium, provide d the original author and source are credited. 78 Bhandari et al. J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 germination (Yang et al., 2011). However, synthetic fungicides are used as the conventional ways of r educing postha r vest r ots which ha ve ma ny drawbacks including high cost, handling hazards, concern about pesticide residue on fruit and a threat to human health and environment (Tzortzakis, 2009). Various synthetic fungicides were identified as toxic and carcinogenic by various researchers (Rouabhi, 2010; Singh et al., 2016). Pathogens also developed r esista nce a ga inst extensive use of synthetic fungicides resulting in declining fungicidal efficiency (Fogliata et al., 2000; Hao et al., 2011). The application of essential oil amended coatings has been developed as a novel and eco-friendly approach to control postharvest microbes, maintain fruit quality and improve shelf life (Alam et al.,2017; Jhalegar et al., 2015). The essential oils do not have only anti- fungal properties, but the secondary metabolites also have antioxidant and bio-regulatory properties (Du Plooy et al., 2009; Jhalegaret al., 2014; Bagamboula et al., 2004; Hendel et al., 2016). The increasing demand for organic fruits encourages replacing synthetic fungicides with safer alternatives. The volatility, ephemeral nature and biodegradability of essential oils make it comparatively advantageous for the treatment of postharvest citrus disease (Ameziane et al., 2007). The synergism between the components in volatiles may be the reason behind the fungitoxic property of essential oils. Therefore, there is a minimal possibility of resistance. The application of essential oils with wax increases its longevity and reduces the amountof essential oils required per fruit. Therefore, the study was made to compare various wax amended treatments, their efficacy and their impact on postharvest parameters. MATERIALS AND METHODS Experimental site and fruit material T he pr esent investiga tion wa s ca r r ied out a t Agriculture and Forestry University (AFU), Rampur, Chitwan during the year 2017-2018. The location of the site is 27o40’ N and 85o19’ E with an elevation of 228 meter above sea level. The experiment was conducted in a cool and humid winter season. The local variety of sweet orange handpicked from the farmers orchard of Sindhuli was transported to Chitwan for the experiment. The fruits were kept in the tagged plastic trays during the storage period at room temperature.The average weight of fruit was 144.56 g. The average seed number was 10. The average juice content of fruit was 84.19 ml. Experimental design T he exper iment wa s la id out in single fa ctor Completely Randomized Design (CRD) with nine treatments and four replications. There were a total of 36 experimental trays having 12 fruits per tray. The treatments were finalized based on the findings of Tripathi et al., (2004), Jhalegar et al., (2015) and Rokaya et al., (2016). Treatments details T1: 10% (w/v) wax emulsion with 0.1% (v/v) Lemongrass (Cymbopogon flexuosus) oil T2: 10% (w/v) wax emulsion with 0.5% (v/v) Lemongrass (Cymbopogon flexuosus) oil T3: 10% (w/v) wax emulsion with 0.1% (v/v) Mentha (Mentha arvensis) oil T4: 10% (w/v) wax emulsion with 0.5% (v/v) Mentha (Mentha arvensis) oil T5: 10% (w/v) wax emulsion with 0.1% (v/v) Eucalyptus (Eucalyptus sp.) oil T6: 10% (w/v) wax emulsion with 0.5% (v/v) Eucalyptus (Eucalyptus sp.) oil T7: 10% (w/v) wax emulsion with 0.1% (v/v) Carbendazim (Bavistin) T8: 10% (w/v) wax emulsion with 0.5% (v/v) Carbendazim (Bavistin) T9: Control (dipped in 10% wax emulsion only) Preparation of 10% wax emulsion Paraffin wax (58-60°C, Solid LR-Grade) was used for preparing wax emulsion. Five hundred milliliter of water was boiled in a vessel and 50 g of wax was hea ted in a nother vessel. Fifteen milliliter of triethanolamine and ten milliliter of oleic acid was added in water as emulsifier and stabilizers. The molten wax was gradually poured into heated water with constant stirring. The stirring was rigorously done until the solution turns milky color. The milky color indicates well pr epa red emulsion. It was ensured that the heated wax and heated water were at same temperature while mixing. The prepared emulsion was then allowed to cool. Preparation and application of essential oils and fungicide The essential oils used in the experiments were pr epa r ed a t Herbs Pr oduction a nd Pr ocessing Cor por a tion Limited (HPPCL), Koteshwor, Kathmandu. The respective herbs collected from the Terai region of Nepal were dried, wilted and steam 79 Fungicide and essential oils amended wax coating for extended shelf of sweet orange Essential oils Chemical constituents Cymbopogan Flexuosus Geranial, Neral, Limonene, Caryophyllene, Geranyl acetate, Linalyl acetate, Citral, Isogeranial, p-cymene, Linalool Mentha arvensis Menthol, Menthone, Isomenthone, Menthyl acetate, Limonene Eucalyptus sp. Eucalyptol, Limonene, Aromadendrene, Phellamdrene, Terpinolene, Alpha terpineol Source: HPPCL website Table 1: Chemical constituents of essential oils used in experiment J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 distilled to produce oils. One milliliter and five milliliter of essential oils were added in one liter of 10% wax emulsion to prepare 0.1% and 0.5% essential oils with wax emulsion. Similarly, the fungicidal solution of carbendazim was prepared by dissolving 1 g and 5 g of carbendazim (Carbendazim 50% WP) in 1000 ml of distilled water. One milliliten and five millititen of fungicidal solution were added in one liter of 10% wax emulsion to prepare 0.1% and 0.5% carbendazim with wax emulsion. The fruits were then dipped in the designated solutions for a few seconds, until a glossy film of wax was formed on the surface of fruits. Data collection and analysis The data about the Juice content, fruit firmness, Total Soluble Solids (TSS), Titratable Acidity (TA), vitamin C (ascorbic acid) and disease severity scoring was taken at every 5th day interval. The physical (fruit firmness) and chemical (juice recovery percentage, TSS, TA and vitamin C) properties of fruits were measured by destructive sampling technique. The fruit firmness was measured bypenetrometer (effigy oil model having 8 mm tip) and TSS was measured by hand refractometer. Acidity and vitamin C was determined as per the procedure outlined by AOAC (2005). The juice recovery percentage and content was calculated by following formulae; The shelf life was evaluated based on the appearance and spoilage of fruits. Fruits were considered to have reached the end of shelf life when fruits showed visible signs of decay irrespective of diameter of symptom (Obagwu and Korsten, 2003). Disease scoring and identification Disea se scor ing wa s done on 0-5 sca le. T he assessment was based on the rotted area with respect to tota l sur face ar ea of the sweet or ange a nd expressed in percentage [0 = no infection (fruits are healthy), 1 = infection starts (0-5% rotting), 2 = 6- 10% rotting, 3 = 11-15% rotting, 4 =16-20% rotting, 5=>20% rotting] (Obagwu and Korsten, 2003; Abd-El-Khair and Hafez, 2006). The rotted fruits from each replication were removed and counted. Disease severity index of decay fruits by pathogen was calculated by following formulae; The infected fruits after treatment with fungicide and essential oils were transferred to the pathology lab of AFU for the isolation of fungi. Isolation was carried out on Martin’s medium (Bridson,1995). Small pieces (1-1.2 cm thickness) of rotted fruits were sterilized by dipping into 2% sodium hypochlorite solution for 5 minutes and then washed several times with distilled water and finally dried on sterile filter paper (Abd- El-Khair and El-Mougy, 2003). The fully sterilized pieces were then transferred onto the surface of the medium in sterilized Petri-plates. Inoculated plates were incubated at 25oC for 3-5 days. Hyphal tip technique was followed for purification of the isolated fungi. Barnett and Hunter technique was used to identify funga l cultur es ( Ba r nett a nd Hunter, 1987). T he temper a tur e and r ela tive humidity of the ex p er imenta l r oom wa s r ec or ded da ily. T he a ver a ge minimum temper a t u r e wa s r ecor ded 12.88°C while the average maximum temperature wa s r ecorded 16. 16 °C. T he a ver a ge minimum humidity was 86.04% while the average maximum Disease severity index (%) = (Abd-El-Khair and Hafez, 2006) Where, n = number of decayed fruits per category, r 1. r5= severity s core M = maximum rating s cale number (5), N = total examined fruits 80 J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Bhandari et al. humidity was 91.73%. The climate was mostly cloudy during the experiment with a few instances of drizzles. The data were entered into Microsoft Excel 2016 and analysis was carried out by using R- Studio version 4.0.2. Both descr iptive and inferential analysis was carried out. Interpretations were made based on results, which were assisted by qualitative and quantitative data/information. RESULTS Juice recovery percentage Juice recovery percentage decreased significantly in all treatments with the advancement of storage time (Table 2). On the day of the experimental setup, the juice recovery percentage was found to be 58.24%. The juice recovery percentage was not significant between treatments for the stor age period time of 5 days and 10 days. At 30 days after storage, maximum juice recovery percentage was observed in wax coating with 0.5% lemongrass (42.86%), which was statistically at par with the wax coating with 0.5% eucalyptus (42.81%). The lowest juice r ecover y per centa ge wa s seen in control fruits (33.49%). Table 2: Effect of postharvest treatments on juice recovery percentage of sweet orange fruits Treatments Per cent juice content of fruits on days indicated 1 5 10 15 20 25 30 T1 58.24 57.04 50.17 47.04abc 45.05c 43.82bc 41.45cd T2 58.24 56.95 49.56 47.81a 46.21a 44.10ab 42.86a T3 58.24 56.73 49.60 46.67bc 45.30bc 43.69c 41.18c T4 58.24 57.38 51.70 47.92a 45.93ab 44.25a 41.63c T5 58.24 57.10 50.60 47.60ab 45.46abc 43.64c 42.29b T6 58.24 57.23 50.82 47.55ab 46.16a 44.10ab 42.81a T7 58.24 56.98 49.80 46.42c 43.50d 39.71e 38.50e T8 58.24 57.20 51.20 46.12c 44.10d 42.02d 41.16d T9 58.24 56.59 50.63 46.08c 40.67e 36.10f 33.49f LSD 0.60ns 0.85ns 1.06** 0.70*** 0.31*** 0.28*** CV 0.73 1.47 1.55 1.08 0.51 0.47 Mean 57.01 50.45 47.02 44.71 42.38 40.59 LSD = Least Significant Difference, CV= Coefficient of Variation, Means within the column followed by same letters do not differ significantly at 5% level of significance by DMRT, Significance codes ***at 0.001, **at 0.01, *at 0.05 Fruit firmness The fruit firmness decreased with the advancement of the storage period in all treatments (Fig. 1). On the day of the experimental setup, the fruit firmness was found to be 5.35 kg/cm2. On the 30th day after storage, firmness was highest for wax with 0.5% eucalyptus (3.50 kg/cm2) and lowest in control (2.25 kg/cm2) followed by wax with 0.1% carbendazim (2.75 kg/cm2). 81 Fig.1 : Effect of postharvest treatments on firmness of sweet orange fruits J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Fungicide and essential oils amended wax coating for extended shelf of sweet orange Total soluble solids (TSS) Total soluble solid directly influences the taste of sweet orange. The TSS of fruit on the first day of storage was 11.20 °Brix. TSS increased with increment in the storage period in all treatments from 10 days onwards (Table 3). However, TSS was found to decrease on the 5th day of storage for the treatment wax with mentha. There was no significant difference between treatments on the 5th and 10th day of storage. The highest TSS was observed in control fruits (12.41° Brix) followed by wax with 0.1% carbendazim (12.28° Brix) while the lowest TSS was observed in wax with 0.5% lemongrass (11.97° Brix) at 30th day after storage. Titratable Acidity (TA) The titratable acidity is an important factor that is directly related to organic acid present in the fruit and also determines the quality of sweet orange. The TA w as 1 .1 2 o n t he first da y o f t he experiment. The effect was significant only after the 10th day of treatment. There was a gradual decrease in TA of sweet orange along with the storage time. At the end of storage life i.e. 30th d a y, TA w a s hig he st fo r w a x w it h 0 . 5 % lemongrass (0.94%), which was statistically at par to wax with 0.1% lemongrass (0.92%) and 0.5% carbendazim (0.91%). The lowest TA was shown by control (0.72%) at the end of storage life (Table 4). 82 Table 4: Effect of postharvest treatments on TA of sweet orange fruits Treatments TA on days indicated 1 5 10 15 20 25 30 T1 1.12 1.05 1.03bc 1.02ab 0.95ab 0.93ab 0.92ab T2 1.12 1.04 1.05b 1.03ab 0.99a 0.95a 0.94a T3 1.12 1.03 0.95de 0.92de 0.91c 0.84d 0.84d T4 1.12 1.04 0.99d 0.94d 0.93bc 0.88c 0.82d T5 1.12 1.05 0.95de 0.9e 0.86d 0.85d 0.82d T6 1.12 1.02 1.04bc 0.98c 0.95abc 0.91b 0.86cd T7 1.12 1.02 1.05bc 1.01b 0.96ab 0.93ab 0.87bcd T8 1.12 1.03 1.08a 1.04a 0.97ab 0.93ab 0.91abc T9 1.12 1.03 0.89e 0.85f 0.76e 0.74e 0.72e LSD 0.00ns 0.12** 0.01*** 0.03*** 0.02*** 0.05*** CV 4.86 1.85 1.30 2.75 2.01 4.00 Mean 1.03 1.00 0.96 0.92 0.88 0.85 LSD = Least Significant Difference, CV= Coefficient of Variation, Means within the column followed by same letters don not differ significantly at 5% level of significance by DMRT, Significance codes ***at 0.001, **at 0.01, *at 0.05. Bhandari et al. Treatments TSS of fruits on days indicated 1 5 10 15 20 25 30 T1 11.20 11.25 11.30 11.40bcd 11.69b 11.82e 12.21c T2 11.20 11.25 11.30 11.40bcd 11.60c 11.72f 11.97d T3 11.20 11.00 11.20 11.41abc 11.65b 11.97bc 12.22bc T4 11.20 11.00 11.30 11.45ab 11.78a 11.8e 12.19c T5 11.20 11.25 11.30 11.37cd 11.65bc 11.9d 12.20c T6 11.20 11.25 11.30 11.40bcd 11.61c 11.8d 12.10d T7 11.20 11.20 11.25 11.34d 11.65bc 12.02b 12.28b T8 11.20 11.25 11.30 11.37cd 11.61c 11.95c 12.19c T9 11.20 11.25 11.30 11.47a 11.78a 12.19a 12.41a LSD 0.20ns 0.08ns 0.06** 0.04*** 0.04*** 0.06*** CV 1.82 0.53 0.37 0.27 0.23 0.35 Mean 11.18 11.28 11.40 11.66 11.91 12.20 LSD = Least Significant Difference, CV= Coefficient of Variation, Means within the column followed by same letters do not differ significantly at 5% level of significance by DMRT, Significance codes ***at 0.001, **at 0.01, *at 0.05. Table 3: Effect of postharvest treatments on TSS of sweet orange fruits J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 83 J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Fungicide and essential oils amended wax coating for extended shelf of sweet orange Ascorbic Acid (Vitamin C) content Vit a min C c ont ent is a n imp or t a nt nu t r it ive parameter in citrus fruits and it was decreased gradually during the advancement of storage days (Fig. 2). On the first day of storage, the vitamin C content was measured to be 40 mg/100ml of orange juice. On the 30th day, the highest vitamin C was f ou nd in f r u it s c oa t ed wit h wa x a nd 0 . 5 % eucalyptus (30.31 mg/100ml), followed by wax with 0.5% mentha (29.50 mg/100ml) and 0.1% mentha (29.18mg/100ml), while the lowest vitamin C was observed in control fruits (24.5 mg/100ml). Disease severity index T he disea se oc cur r ence in sweet or a nge wa s increased with the storage days (Table 5). The green mold (P. digitatum) was confirmed through the lab culture of a pathogen. The fruits treated with essential oils and fungicide were found to be more resistant to postharvest fungal diseases. On the 30 th day of stor a ge, a lmost a ll trea tments exhibited noticeable disease occurrence, control being the highest infected (0.372%) followed by wax with 0.1% mentha (0.180%). The treatment of wax with 0. 5% car bendazim (0. 004%) wa s most effective against fungal pathogen and wax with 0.5% euca lyptus oil (0. 025%) and 0. 5% lemongrass oil (0.025%) being the most effective essential oils. Fig. 2: Effect of postharvest treatments on Ascorbic Acid content of sweet orange fruits 84 Treatments Disease severity index of fruits on days indicated 1 5 10 15 20 25 30 T1 0.00 0.00 0.00 0.00 0.025bc 0.075b 0.123b T2 0.00 0.00 0.00 0.00 0.004de 0.012cd 0.025d T3 0.00 0.00 0.004b 0.012b 0.038b 0.075b 0.180b T4 0.00 0.00 0.00 0.00 0.017cd 0.046bc 0.114bc T5 0.00 0.00 0.00 0.012b 0.038b 0.058b 0.114bc T6 0.00 0.00 0.00 0.00 0.008de 0.012cd 0.025d T7 0.00 0.00 0.00 0.00 0.00 0.000 0.033cd T8 0.00 0.00 0.00 0.00 0.00 0.000 0.004d T9 0.00 0.00 0.042a 0.054a 0.096a 0.207a 0.372a LSD 0.009*** 0.013*** 0.013*** 0.038*** 0.080*** CV 121.76 105.94 35.646 48.471 50.13 Mean 0.005 0.008 0.025 0.054 0.110 LSD = Least Significant Difference, CV= Coefficient of Variation, Means within the column followed by same letters do not differ significantly at 5% level of significance by DMRT, Significance codes ***at 0.001, **at 0.01, *at 0.05. Table 5: Effect of postharvest treatments on disease severity index in sweet orange fruits J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Bhandari et al. Shelf life The wax treatment with carbendazim and essential oils had a significantly better shelf life as compared to the control treatment (Table 6). Wax with 0.5% carbendazim (28.25 days) being the highest and significantly better than other treatments. It was followed by wax with 0.1% carbendazim (25.75 days), wax with 0.5% lemongrass oil (20.00 days) and wax with 0.5% eucalyptus oil (19.75 days). The control fruits (8.25 days) were observed to have the lowest shelf life. Discussion A significantly lower juice recovery percentage of control fruits might be due to the fact that the essential oils act as a barrier which checks the loss of moisture from the fruit surface due to the clogging of na tur a l openings (Ca stillo et al. , 2014). Additiona lly, the lower incidence of disease in essential oils and fungicides treated fruit ensure lower metabolism, which might have contributed to a higher juice recovery percentage. The present finding was supported by (Bisen et al., 2012). The control fruit also had a wax coating and the transpiration process wa s ver y slow, so ther e wa s a n insignifica nt difference in juice recovery percentage between treatments before 15th day of storage. The moisture Table 6: Effect of postharvest treatments on shelf life in sweet orange fruits LSD = Least Significant Difference, CV= Coefficient of Variation, Means within the column followed by same letters don not differ significantly at 5% level of significance by DMRT, Significance codes ***at 0.001, **at 0.01, *at 0.05. Treatments Shelf life T1 17.25d T2 20.00c T3 10.00f T4 16.75d T5 14.25e T6 19.75c T7 25.75b T8 28.25a T9 8.25g LSD 0.92*** CV 3.55 Mean 17.80 85 J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Fungicide and essential oils amended wax coating for extended shelf of sweet orange loss was found significantly lower in fruit treated with essential oil enriched coatings (Du Plooy et al., 2009; D Antunes et al., 2012; Castillo et al., 2014). In general, coating formulations that minimize weight loss are also better at maintaining firmness, since this attribute is highly influenced by water content. Fruit firmness decreased gradually and significantly along with increasing storage period in all treatments. The decelerated damage may be due to the anti-microbial properties of essential oils. The lowest fruit firmness in control fruit might be due to the rapid degradation of cell walls due to the action of wall-degrading enzymes such as pectinestera se, pectinmethyl­ esterase and polygalacturonase which are produced by fungi. Essential oil amended coating maintains cell wall carbohydrate metabolism during storage which is related to decreased susceptibility to infection by fungal pathogen and therefore improves quality. The essentials oils together with commercial wax coating maintain the orga noleptic integrity along with firmness as mentioned by Jhalegar et al. (2014). The essential oils affect the portioning of the lipids of the plasma membrane and changing of its integrity, permeability and inorganic ion equilibrium due to their hydrophobic nature (Lambert et al., 2001) which might be the reasons for greater firmness in the fruits treated essential oils. The present findings were supported by Chafer et al. (2012) on the firmness of Navel Powell orange and Castillo et al. (2014) on lemon fruits. The gradual increase of TSS with extending of the storage period might be attributed to concentrated juice content results from dehydration and hydrolysis of polysaccharides.The increased respiration rate due to microbial spoilage, degradation of fruits and increased ethylene production ultimately increased the TSS during ripening and senescence which might be the reason for slightly higher TSS in control fruits as compared to other treatments. The present result was in agreement with the findings of Chafer et al. (2012) on Navel Powell orange and Castillo et al. (2014) on lemon and Tao et al. (2014) on Satusma ma nda rin, as the essentia l oils did not show a significant effect on TSS. The present finding was also inconsistent with Asghari et al. (2009) who reported insignificant results in TSS while using cumin essential oil on strawberry. The decrease in titratable acidity with storage is due to the oxida tion of orga nic a cids a nd fur ther utilization in the metabolic process in the fruits (Hafeez et al.,2012). A gradual declining trend in titratable acidity content of fruit dur ing stor age for a ny treatment was observed by Ansari and Feridoon (2007) and Obenland et al. (2008) in citrus. The decreased in titratable acidity of fruits during storage could be due to the consumption of organic acids in the respiration process as stated by Zokaei et al. (2006) and Ishaq et al. (2009). Similarly, Baiea (2013) on Washington Navel orange detected a decrease in the acidity of fruits during storage. Fruits treated with essential oils showed higher retention of titratable acidity during the storage period which might due to delayed in physiological ageing and alteration in metabolism. The present results are in line with Mahajan et al. (2010) suggesting that organic acids were used in the respiratory process. The higher titratable acidity in wax with lemongrass treatment is aligned with the finding of Fatemi et al. (2012) who reported that the thymol oil delayed the changes in titratable acidity of Valencia orange. The present finding was also supported by Jhalegar et al. (2014) on Kinnow mandarin. Abd El wahab et al. (2014) also reported bergamot oil delayed the changes in titratable acidity during cold storage of Crimson seedless grape. Adisa (1986) stated that vitamin C decreased over time in storage which is similar to the experimental outcome. The decreased in ascorbic acid content of fruits during storage could be due to the conversion of dehydroascorbic to diketogulonic acid by oxidation as reported by Ishaq et al. (2009). Under stress, such as a pathogen or chemical exposure, a scorbate oxidase levels were increased, which decreased the level of vitamin C (Loewus and Loewus,1983; Loewus et al.,1987). The maximum retention of vitamin C was observed with essential oils treatments due to the antioxidant property of essential oils which prevent ascorbic acid from oxidation (Shao et al., 2013). The result was similar to Lin et al. (2011) who found that the decrease in vitamin C level was associated with a reduced capacity of preventing oxidative damage which is triggered by the incidence of physiological disorders during storage. The degradation of vitamin C was highest in control fruits which might be due to fruit senescence accompanied by rapid respiration, ethylene production and decay. These results are similar to those reported on the effect of thyme and 86 Table 7: Summary of studies on the effect of essential oils on major post-harvest pathogens of citrus Fruit Target pathogen Essential oils References Orange cv. Tomango P. digitatum Mentha oil Du Plooy et al. (2009) Orange, Lime P. italicum Mentha oil Tripathi et al. (2004) Washington Navel Orange P. digitatum Lemongrass oil, Abd-El-Khair and Hafez Eucalyptus oil (2006) Valencia Orange G. citri-aurantii Lemongrass oil Regnier et al. (2014) Kinnow Mandarin P. digitatum, P. italicum Eucalyptus oil Jhalegar et al. (2014) Kinnow Mandarin P. italicum and Lemongrass oil, Jhalegar et al. (2015) P. digitatum Eucalyptus oil J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Bhandari et al. clove oil in maintaining ascorbic acid as for orange (Zeng et al., 2012; Baiea and Ei-Badawy, 2013). The result was similar to the report of Abd-El- Kha ir and Ha fez (2006), a s they r eported the lemongr a s s a nd eu c a lyp t u s es s e nt ia l oils significantly reduced the incidence of fungus P. digitatum in Wa shington na vel or a nge dur ing storage. Abdolahi et al., (2010) and Al-Samarrai et al., (2013) found various plant extracts including lemongr ass extr a ct could inhibit the mycelia l growth of pathogenic fungus P. digitatum. The phenolic c omp ounds a nd t heir der iva t ives of essential oils altered the microbial cell permeability by interacting with membrane proteins which would cause deformation in cell structure and function and permit the loss of macromolecules from their body (Fung et al., 1997; Rattanapitigorn et al., 2 0 0 6) whic h might b e t he r ea s ons of lower microbial growth in the essential oils treated fruits c omp a r e t o c ont r ol. Amit a nd M a lik ( 2 0 10 ) indic a t ed t ha t t he va p ou r s of lemongr a s s oil da ma ged t h e c ell memb r a ne ma i nly du e t o membrane deformation. However, the variation in the antifungal effect of the essential oils depends on the solubility and capacity to interact with the cytoplasmic membrane (Tripathi and Shukla, 2007). T he ef f ic a c y of lemongr a s s wa s a ls o f ou nd superior by Jhalegar et al. (2015). Similar results were reported by Du Plooy et al. (2009), Fan et al. (2014), Jhalegar et al. (2014) and Gandarilla- Pacheco et al. (2020) in citrus fruits. The wax with 0.5% carbendazim with its prominent disease resistance had the longest storability. The superiority of shelf life of 0.5% lemongrass and eucalyptus oils treated fruits might be due to the antifungal properties of essential oils (Tzortzakis a nd Economa kis, 2007; Jha lega r et al. , 2014; J ha le ga r e t a l . , 2 0 1 5 ) . I n a ddit ion t o t his , postha rvest decay is positively correlated with ethylene production and respiration rate which were f ou nd t o be decr ea sed b y the a p p lica tion of essential oils (Jhalegar et al., 2014; Jhalegar et al., 2 0 1 5 ) . T he p r es ent r es u lt wa s i nc ons is t ent with Tripathi et al. (2004) and Tavakoli et al. (2019). The green mold is the major postharvest pathogen in sweet orange. The chemical fungicides have been found effective against such pathogens, but the health hazard of such pesticides is alarmingly high. The use of essential oils as an alternative for chemicals can be an environment friendly technique for prevention of the health hazards. The shelf life of sweet orange can be extended by infusing wax wit h ca r b enda z im or es s ent ia l oils . Bu t t he superiority of essential oils especially wax with 0. 5% eu ca lyptus oil a nd 0. 5% lemongr a ss in qualitative parameters as well as in consumer ’s p r ef er enc e s , or ga nic r equ ir e ment s a nd envir onmen t a l a s p ec t s ma ke t he m a b et t er alternative. 87 J. Hortl. Sci. Vol. 16(1) : 77-90, 2021 Fungicide and essential oils amended wax coating for extended shelf of sweet orange Abd El Wahab, W.A., Abd El Wahab S.M., Kamel, O. T. 2014. Using sa fe a lter na tives for controlling postharvest decay, maintaining quality of Crimson seedless grape. World Applied Sciences Journal, 31(7):1345-1357. Abd-El-Khair, H. and El-Mougy, N. S. 2003. 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