Postharvest Biology and Technology 23 (2001) 167 170 www.elsevier.

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Short communication

Internal browning in cold-stored pineapples is suppressed by a postharvest application of 1-methylcyclopropene

S. Selvarajah *, A.D. Bauchot, P. John
Department of Agricultural Botany, School of Plant Sciences, The Uni6ersity of Reading, Reading RG6 6AS, UK Received 25 October 2000; accepted 13 February 2001

Abstract Treatment with 1-methylcyclopropene (1-MCP), the inhibitor of the ethylene receptor, at 0.1 ppm (4.5 nmol l 1) for 18 h at 20C effectively controlled internal browning, a chilling injury symptom, in pineapples stored at 10C for four weeks. The treatment with 1-MCP also delayed ascorbic acid decline, and arrested the decline in both total soluble solids and ethylene synthesis. The present ndings throw light on the role of ethylene in internal browning, and suggest that 1-MCP could be considered for use commercially to control this important postharvest physiological disorder in pineapples. 2001 Elsevier Science B.V. All rights reserved.
Keywords: 1-Methylcyclopropene; Ananas comosus L.; Ethylene; Internal browning; Chilling injury

1. Introduction Internal browning (IB, also known as endogenous brown spot or black heart) is the most important physiological disorder of pineapples that are stored below 13C (Dull, 1971), limiting both the storage and the export of this fruit. Partial control only has been achieved after harvest by various treatments (Paull and Rohrbach, 1985; Selvarajah and Herath, 1997; Selvarajah et al., 1997, 1998).
* Corresponding author. Tel.: + 44-118-9318098; fax: + 44118-9316577. E-mail address: selvashanthi@yahoo.com (S. Selvarajah).

1-Methylcyclopropene (1-MCP) is an inhibitor of ethylene perception that binds irreversibly to the ethylene-binding protein (Sisler and Serek, 1997). Since it is non-toxic (Technical Bulletin, Rohm and Haas Company) and odourless, 1MCP is potentially of commercial value to control ethylene-dependent postharvest disorders. Direct involvement of ethylene in pineapple IB has not been reported, but chilling injury is known to be associated with ethylene synthesis (Ben-Amor et al., 1999), even in non-climacteric fruit (McCollum and McDonald, 1991). Thus there appeared to be a basis for testing the effectiveness of 1MCP in controlling IB in pineapple. Here we report that 1-MCP strongly reduces the incidence of IB in pineapple.

0925-5214/01/$ - see front matter 2001 Elsevier Science B.V. All rights reserved. PII: S 0 9 2 5 - 5 2 1 4 ( 0 1 ) 0 0 0 9 9 - 0

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2. Materials and methods Queen pineapples (Ananas comosus. L) from South Africa were transported within three days to our laboratory. Fruit of uniform shape, size, colour and weight were selected and placed individually in 6.7 l sealed plastic containers. One set of fruits was exposed to 0.1 ppm (4.5 nmol l 1) 1-MCP generated by EthylBloc at 20C for 18 h (8090% RH). Control fruit were subjected to the same conditions without exposure to 1-MCP. Following treatment, all the pineapples (treated and control) were stored at 10C (70 80% RH). Fruits were analysed for IB intensity, ripeness stage and other quality parameters immediately upon arrival and then after 1 4 weeks of storage at 10C followed by 3 days shelf-life at 20C (60 70% RH). For each assessment, 10 fruit treated with 1-MCP were compared with 10 control fruit. The trial was repeated twice, in early May and at the end of June, and the results were pooled. The fruit were cut longitudinally in half and the incidence of IB was determined. For each fruit, IB intensity was scored from 0 to 5 according to the percentage of esh affected (0, free from IB; 0.5, watery spots; 1-5: B10, 10 25, 25 50, 50 75 and \75% of the esh discoloured, respectively; Teisson, 1979). The average IB intensity was calculated for each lot of fruit. The stage of ripeness was determined by visual assessment of the shell (Rangana 1977). The scale ranges from 0 to 5: 0, all eyes are totally green; 1, B20% of the eyes are predominantly yellow; 2, 2040% of the eyes are tinged with yellow; 3, up to 65% of the eyes are predominantly yellow; 4, 6590% of the eyes are fully yellow; 5, \90% of the eyes are fully yellow and no more than 20% of the eyes are reddish orange. Fruit with \20% of the eyes reddish orange were considered as senescent and discarded. Each fruit was weighed upon arrival and after storage at 10C followed by 3 days shelf-life at 20C. The total soluble solids (TSS, expressed as % Brix) of fruit juice was determined using a hand-held refractometer (0 30% Brix). Ascorbic acid was determined by HPLC (Slack, 1987). Ethylene emission was monitored by incubating indi-

vidual fruits in plastic containers at 10C and was measured by gas chromatography after a 2 h incubation.

3. Results After only 1 week of storage at 10C followed by 3 days shelf-life, 50% of the control pineapples showed IB, and after 3 weeks storage all were severely affected (Fig. 1A). Treatment with 1-

Fig. 1. Effect of 1-MCP on internal browning incidence (A) and intensity (B), and ripeness (C) of pineapples stored at 10C followed by 3 days at 20C. Asterisks indicate a signicant difference between the 1-MCP treatment and the control at PB 0.01.

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Fig. 2. Effect of 1-MCP on ethylene production of pineapples during storage at 10C. Vertical lines represent standard error of the means and are not shown when the values are smaller than the symbol.

MCP completely eliminated IB for the rst 3 weeks of cold storage, and reduced the incidence to 20% at week 4 (Fig. 1A) with an even more marked effect on the intensity of browning (Fig. 1B). Treatment with 1-MCP also delayed shell ripening, as measured by the eye colour (Fig. 1C). In the control fruit, 65% of the eyes turned yellow (scored as index 3) within 2 weeks, while it took 4 weeks for the 1-MCP treated fruit to reach this stage. The 1-MCP treated fruit did not begin to ripen until more than 2 weeks in storage. Control cold-stored pineapples showed a rapid decline in the rate of ethylene synthesis. The decline in rate of ethylene synthesis was also signicantly slowed by the 1-MCP treatment (Fig. 2). The rapid rate of ascorbic acid decline in pineapples was signicantly delayed by the 1MCP treatment (Fig. 3A). The ascorbic acid levels in the control fruit after one week were reached only after four weeks of storage in 1-MCP treated fruit (Fig. 3A). Treatment with 1-MCP also arrested the decline in TSS (Fig. 3B) and limited the weight loss compared to the control fruit (P B 0.01, t-test; data not shown).

(Sisler and Serek, 1997). Thus, we assume that the effects of 1-MCP reported here are due to 1-MCP blocking the ethylene receptors of pineapple. Although IB in pineapple has not previously been directly demonstrated to be due to ethylene, the present results clearly show that ethylene perception is a necessary step for pineapple to develop chilling injury symptoms. 1-MCP has already been shown to dramatically reduce postharvest chilling injury in climacteric fruits such as melon (Ben-Amor et al., 1999) and apple (Rupasinghe et al., 2000; Watkins et al., 2000), but was not effective on non-climacteric orange (Porat et al., 1999). Besides inhibiting IB development, exposure to 1-MCP stimulated ethylene production in coldstored pineapples. Sisler and Blankenship (1993) already reported a greater capacity for ethylene production when the ethylene receptors were inactivated in mung bean seedlings. Moreover, stimulation of the receptor by ethylene treatments led

4. Discussion It is widely accepted that 1-MCP binds to the ethylene receptors, and blocks ethylene perception

Fig. 3. Effect of 1-MCP on the content of ascorbic acid (A) and total soluble solids (B) of pineapples stored at 10C followed by 3 days at 20C. Vertical lines represent standard error of the means and are not shown when the values are smaller than the symbol.

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S. Sel6arajah et al. / Posthar6est Biology and Technology 23 (2001) 167170 Paull, R.E., Rohrbach, K.G., 1985. Symptom development of chilling injury in pineapple fruit. J. Am. Soc. Hortic. Sci. 110, 100 105. Porat, R., Weiss, B., Cohen, L., Daus, A., Goren, R., Droby, S., 1999. Effects of ethylene and 1-methylcyclopropene on the postharvest qualities of Shamouti oranges. Postharvest Biol. Technol. 15, 155 163. Rangana, S. (Ed.), 1977. Manual of analysis of fruits and vegetable products. Tata McGraw-Hill, New Delhi, pp. 94 95. Rupasinghe, H.P.V., Murr, D.P., Paliyath, G., Skog, L., 2000. Inhibitory effect of 1-MCP on ripening and supercial scald development in McIntosh and Delicious apples. J. Hortic. Sci. Biotechnol. 75, 271 276. Selvarajah, S., Herath, H.M.W., 1997. Effect of edible coating on some quality and physico-chemical parameters of pineapple during cold storage. Trop. Agric. Res. 9, 77 89. Selvarajah, S., Herath, H.M.W., Liyanage, K.K., 1997. Effect of modied atmospheric storage on internal browning development in pineapple. Trop. Agric. Res. 9, 388 391. Selvarajah, S., Herath, H.M.W., Bandara, D.C., Abeysinghe Banda, D.M.G., 1998. Effect of pre-harvest calcium treatment on post-harvest quality of pineapple. Trop. Agric. Res. 10, 214 224. Sisler, E.C., Blankenship, S.M., 1993. Diazocyclopentadiene (DACP), a light sensitive reagent for the ethylene receptor in plants. Plant Growth Regul. 12, 125 132. Sisler, E.C., Serek, M., 1997. Inhibitors of ethylene responses in plants at the receptor level. Recent developments. Physiol. Plant. 100, 577 582. Slack, P.T. (Ed.), 1987. Analytical Methods Manual, 2nd ed. British Food Manufacturing Industries Research Association, Leatherhead, UK. Teisson, C., 1979. Internal browning of pineapples: I. History; II. Materials and Methods. Fruits 34, 245 261. Vendrell, M., McGlasson, W.B., 1971. Inhibition of ethylene production in banana fruit tissue by ethylene treatment. Aust. J. Biol. Sci. 24, 885 895. Watkins, C.B., Nock, J.F., Whitaker, B.D., 2000. Responses of early, mid and late season apple cultivars to postharvest application of 1-methylcyclopropene (1-MCP) under air and controlled atmosphere storage conditions. Postharvest Biol. Technol. 19, 17 32.

to a decrease in ethylene production in immature banana (Vendrell and McGlasson, 1971). The phenomenon is known as auto-inhibition of ethylene production. It is compatible with our observation that 1-MCP treatment induced ethylene production in pineapple. In addition, 1-MCP delayed shell yellowing by two weeks, as previously observed for cold-stored orange peel (Porat et al., 1999). This conrms the involvement of ethylene perception in non-climacteric fruit degreening. It also suggests that 1-MCP inhibited ethylene perception for between two and three weeks in pineapple. In conclusion, the suppression of IB demonstrated here suggests that 1-MCP could be considered for commercial application in controlling this widespread disorder during the storage and transport of pineapples.

Acknowledgements We thank Safeway for providing fruits and Rohm and Haas Company for kindly providing EthylBloc. We are grateful to Sue Mitchell and Jennifer Greenham for expert technical advice.

References
Ben-Amor, M., Flores, B., Latche, A., Bouzayen, M., Pech, J.C., Romojaro, F., 1999. Inhibition of ethylene biosynthesis by antisense ACC oxidase RNA prevents chilling injury in Charentais cantaloupe melons. Plant Cell Environ. 22, 1579 1586. McCollum, T.G., McDonald, R.E., 1991. Electrolyte leakage, respiration and ethylene production as indices of chilling injury in grapefruits. Hortscience 26, 1191 1192.