Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Progress Report Understnding the reltionship between florl frgrnce, ethylene production nd vse life of cut rose flowers An Mri Bord Dvid Clrk Terril Nell UNIVERSITY OF FLORIDA Ginesville, Florid Report Dte: August 15, 28 (27-8 Finl Report Funded by the Joseph H. Hill Memoril Foundtion, Inc. ICFG-HILL, P.O. Box 99, Hslett, MI 4884 ICFG.HILL@yhoo.com Pge 1 of 12
Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Summry The three min objectives of this reserch project re: I. Determine the min voltile orgnic compounds tht chrcterize the frgrnce profile of frgrnt nd non frgrnt cut rose cultivrs. II. Describe the frgrnce profile of cut roses during vse life nd the effect of externl nd internl ethylene on the frgrnce profile nd vse life of cut roses. III. Study the effects of nti-ethylene tretments on the frgrnce profile nd vse life of frgrnt nd non-frgrnt cut rose flowers. During the period from Jnury through July, 28 the effects of externl ethylene nd ntiethylene tretments were evluted on ( Allure, Lovely Drem, Erin, Osin nd Red Senstion ). The ethylene synthesis inhibitor (AVG) nd the ethylene sensitivity inhibitor (STS) were evluted to determine whether ethylene synthesis or sensitivity exerts the most control over vse life of cut roses. During the course of these experiments, we evluted the effects of externl ethylene nd nti-ethylene tretments on the frgrnce voltile profile of cut roses. MATERIALS AND METHODS Flowers were hrvested between 8-1 m t the sme dy t commercil stge from frms locted in Ecudor. Flowers were tken into the posthrvest room (14-18 o C) within 3 minutes from hrvest, where they were plced into fresh hydrtion solution (wter, 6 ppm chlorine nd citric cid to djust ph to 4.5) for 2 hours. After hydrtion flowers were processed nd plced into the cooler t 3 o C in the sme solution for 12 hours. The next dy, flowers were pckged into 2 commercil hlf boxes. In order to protect the flowers from n eventul ethylene exposure during shipment, 1-MCP schets (EthylBloc TM, Florlife, Inc., Wlterboro, SC) were included inside one shipping box. Two 1-MCP schets were dipped in wter prior plcement inside one box. The box ws closed immeditely fter the 1-MCP schet plcement. The other box contined no 1-MCP schets. Ech box contined 7 bunches ech with 25 stems ech (5 cm in length). A temperture recorder ws plced inside one box. The boxes were send to the crrier gency nd then vi FedEx to Ginesville, Fl within 3-4 dys. Boxes generlly rrived to Ginesville, Fl between 1m to 1 pm. The boxes were opened nd flowers were processed immeditely fter rrivl. Flowers were re-cut to 45 cm in length nd plced into different hydrtion solutions (Tble 1). Flowers tht received the 1-MCP tretment during shipment were plced in bucket with solution of.2 mm Silver thiosulphte (STS). The STS solution ws prepred following the protocol from Esmeli et l 25. Flowers with no 1-MCP were hydrted either in di wter or 1 ppm minoethoxyvinylglycine (AVG) ( Vlent U.S.A Corportion, Wlnut Creek, CA). The totl volume of hydrtion solution ws 5 liters nd the hydrtion ws performed t 21 o C for 3 hours. After 3 hours, the flowers tht were in STS solution were trnsferred to solution of di wter. Then flowers from ll tretments were plced t 3 o C overnight. Pge 2 of 12
Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Tble 1. Hydrtion solutions nd ethylene tretments performed. Anti-ethylene tretment during shipment Plus 1-MCP Hydrtion fter shipment Hydrtion during externl ethylene exposure Externl ethylene ppliction Hydrtion during vse life, ethylene, CO 2 nd voltile determintion.2mm Di wter 1ppm Di wter STS ir No 1-MCP Di wter Di wter 1ppm Di wter ir 1 ppm 1 ppm AVG 1ppm 1 ppm AVG AVG ir The next dy, flowers were plced into two different vse solutions: di wter (for di nd sts pretreted flowers) or 1 ppm AVG (for AVG pre-treted flowers) (Tble 1). The solutions were kept the sme throughout the vse life, ethylene, CO 2 nd voltile collection evlutions. The vses were plced into seled qurium glss tnks nd exposed to either 1ppm ethylene or ir for 24 hours t 21 o C nd 12 hour photoperiod. After the ethylene/ir exposure flowers were tken out of the tnks nd rndomly distributed for vse life, ethylene, CO 2 nd voltile production determintion. Vse life Flowers selected for vse life evlution were plced into vses with 1 l of either di or AVG solution (Tble 1). Flowers were mintined t 21 o C, 5-6 % RH nd 1 mol m -2 s -1 of light (12 hrs/dy). Flower vse life ws determined s the time from plcement in the vse (dy) to the ppernce of visul senescence symptoms (i.e. petl wilting, bscission, bluing nd disese). Flower opening ws rted t dy 1, 2 nd then every two dys until dy 8 using the following scores: 1 = outer petls tightly wrpped round bud, 2 = outer petls strting to reflex from bud, 3 = outer petls reflexed pproximtely 135 o to stem, 4 = outer petls reflexed t pproximtely 115 o to stem, 5 outer petls reflexed t 9 o to stem (modified from (Kuiper et l., 1996)). Ethylene production nd Respirtion rte Flowers selected for ethylene nd CO 2 production were cut to 6 cm in length nd plced into 22 ml glss vils with di wter (sts nd controls) or AVG (Tble 1). For ethylene production determintion flowers were plced into 9ml glss mson jrs with 8ml of 1 M KOH to mintin CO 2 concentrtion below.1 %. Jrs were closed for 21 hours t 21 o C. Then 1 ml gs smple ws tken from the jr using 1 ml syringe nd nlyzed with FID gs chromtogrph (Hewlett-Pckrd 589 Series II). Injector, detector nd oven tempertures were 11, 15 nd 13 o C respectively. Different flowers were used to determine CO 2 production. In this cse the jrs were closed for 2 hours t 21 o C. Then, 1mL of gs ws tken out of the jr nd nlyzed on Gow-Mc gs chromtogrph (Series 58, Bridge wter, NJ) equipped with therml conductivity detector (TCD). The crrier gs (helium) flow rte ws.5mls 1. The oven ws set t 4 C nd the detector nd injector were operted t mbient conditions. Jrs were then opened nd identicl mesurements were tken the following dy. Once the jrs were opened the fresh Pge 3 of 12
Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 weight of ech flower nd the weigh of the vil were determined. Ethylene nd CO 2 ws mesured during 7 dys. Voltile collection Voltiles were collected fter, 2, 4 nd 6 dys of the ethylene/ir tretment. Flowers were recut to 6 cm in length nd plced into 22 ml glss vils with di wter (di nd STS tretment) or AVG (Tble 1). Then flowers were plced into 9 ml glss jrs tht were connected with continuous ir flow system. Crbon filtered ir ws pumped/pulled through the jrs for one hour nd trpped on sorbent Super Q column to concentrte the voltiles. Voltiles collected on the sorbent were eluted with methylene chloride (Fisher Scientific). Nonyl cette ws dded s stndrd fter collection. Smples were nlyzed by gs chromtogrphy nd mss spectrometry s described by Underwood et l., (25). Voltiles were collected t 21 o C between 3:3 nd 4pm t ech smpling dte. At ech smpling time, flower opening ws rted using the following scores: 1 = outer petls tightly wrpped round bud, 2 = outer petls strting to reflex from bud, 3 = outer petls reflexed pproximtely 135 o to stem, 4 = outer petls reflexed t pproximtely 115 o to stem, 5 outer petls reflexed t 9 o to stem (modified from (Kuiper et l., 1996)). The sme flowers from ech tretment were used during the 6 dy collection. Sttisticl nlysis Three replicte vses contining eight stems ech were used per tretment for vse life evlution. Flowers were rrnged in vses for evlution in completely rndomized block design. Dt were nlyzed by ANOVA using SAS Version 8 (SAS Institute Inc., Cry, NC, USA). The significnce of differences mong tretment dt mens were determined using LDS multiple comprison test t p=.5. Four stems were used per tretment for ethylene detection nd three stems per tretment were used for respirtion determintion. Two flowers were used per tretment for voltile extrctions. Pge 4 of 12
Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 RESULTS Vse life Externl ethylene (1ppm for 24 hours t 21 o C) hd negtive effect on the vse life of Allure, Lovely drem, Erin, Osin nd Red senstion vrieties compred to ir exposure. Osin flowers hydrted in di wter nd treted with ethylene hd the highest externl ethylene sensitivity in terms of reduction in vse life. Vse life of Osin ws reduced in 1 dys with externl ethylene ppliction compred to ir exposure. Allure hd reduction of 3 dys, Red Senstion nd Erin 2 dys nd Lovely Drem 1 dy. The negtive effects of externl ethylene on vse life were prevented when flowers were treted with STS. The inhibition of ethylene perception improved vse life on Allure, Lovely Drem, Erin, Osin nd Red senstion flowers compred to controls. However, these effects were not prevented by the inhibition of ethylene synthesis. AVG did not produce ny ffect on the vse life of Allure, Osin nd Red Senstion compre to controls. However, it improved vse life on Lovely Drem nd Erin flowers compred to controls (Figure 1). Ethylene production Ethylene production depended on the vriety nd on the inhibition of ethylene perception. The highest ethylene production ws observed in Lovely Drem with pek observed fter 2 dys. The production then declined to low levels towrds the 7 dy. Allure, Erin, Osin nd Red Senstion produced significntly lower mounts of ethylene compred to Lovely Drem during the 7 dy period. No ethylene production peks were observed on these vrieties. Ethylene production ws lower thn controls on Lovely Drem flowers with no ethylene perception. On the other hnd, ethylene production ws higher thn controls on flowers of Allure, Erin nd Osin with no ethylene perception. Ethylene production ws very low in Red senstion nd it did not vried with inhibition of ethylene perception (Figure 2). Respirtion rte Respirtion rte depended on the vriety. The inhibition of ethylene perception produced low respirtion rtes on some vrieties. The inhibition of ethylene synthesis produce higher respirtion rte in one cultivr but it did not hd ny effect on the other 4. The highest respirtion rte ws observed on Allure followed by Lovely drem, Osin. Erin nd Red Senstion flowers hydrted in di wter nd treted with ir (Figure 3). Inhibition of ethylene perception decresed the respirtion rte in Lovely Drem, Erin nd Red Senstion fter externl ethylene exposure. The sme trend ws observed in Red Senstion fter 4 nd 6 dys of ethylene exposure. Contrry, the inhibition of ethylene synthesis incresed the respirtion rte in Lovely Drem. Lovely Drem treted with AVG trnspired higher thn control fter 5 dys of externl ethylene exposure. Additionlly, the inhibition of ethylene synthesis hd no effects on the respirtion rte of Allure, Erin, Osin nd Red Senstion (Figure 4). Pge 5 of 12
Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Voltile collection Voltile production chnged over time on ll vrieties. After externl ethylene exposure, totl voltile production decresed over time in Allure, Osin nd Red Senstion flowers hydrted in STS, AVG nd di wter. Contrry, voltile production incresed over time on Lovely Drem nd Erin on flowers treted with STS, AVG nd di wter (Figure 5). Inhibition of ethylene synthesis nd production slightly chnged the voltile production of rose flowers over time. After externl ethylene exposure, AVG treted flowers of Allure, Lovely Drem nd Red Senstion produced less frgrnce thn control immeditely fter externl ethylene exposure. This tend continued over time in Lovely Drem nd Red Senstion. Erin flowers treted with AVG produced higher mount of totl voltiles thn the control immeditely fter ethylene exposure. Then t dy 2 the voltile production ws lower thn the control nd continued low over time. Osin voltile production ws not different from control on AVG treted flowers. After externl ethylene exposure, the totl voltile production on Allure Osin nd Red Senstion flowers treted with STS ws not different from the control over the six dys of mesurements. However, Lovely Drem nd Erin flowers treted with STS nd ethylene produced less frgrnce thn the control t dy, then the voltile production incresed nd ws higher thn controls on dys 2 nd 4. Finlly voltile production ws gin lower thn the control t dy 6 for Lovely Drem nd not different from control on Erin (Figure 6). Pge 6 of 12
vse life (verge dys) Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 1. Vse life of frgrnt nd non frgrnt cut rose cultivrs fter ethylene exposure (1pmm) for 24 hours t 21 o C. Prior to externl ethylene exposure flowers were hydrted in di wter, 2.mM STS or 1 pmm AVG. Mens nd stndrd error of 24 flowers. Different letters = significnt difference between mens =.5. 9 8 7 6 5 4 3 2 1 d d plus ethylene Allure b c No ethylene di sts vg 8 7 6 5 4 3 2 1 c d plus ethylene Lovely Drem b b no ethylene di sts vg 16 14 12 1 8 6 4 2 Erin c b b b Plus ethylene No ethylene di sts vg 14 12 1 8 6 4 2 c c plus ethylene Osin b no ethylene di sts vg Red Senstion 12 1 8 6 4 b b di sts vg 2 plus ethylene no ethylene Pge 7 of 12
ethylene production (ul/kg/h) ethylene production (ul/kg/h) ethylene production (ul/kg/h) ethylene production (ul/kg/h) ethylene production (ul/kg/h) Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 2. Ethylene production of cut rose cultivrs. Flowers were hydrted in di wter, 2.mM STS or 1 pmm AVG. Mens nd stndrd error of 4 flowers. 1. 8. 6. 4. 2.. ex 16 Allure No ethylene ir di ir sts ir vg 14. 12. 1. 8. 6. 4. 2.. Lovely Drem ir di ir sts ir vg Erin ir di 3. ir sts ir vg 2. 1.. 8 2. 1.5 1..5. Osin ir di ir sts ir vg 2. 1.5 1..5 Red Senstion ir di ir sts ir vg. Pge 8 of 12
respirtion rte (ml/kg/h) Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 3. Respirtion rte of cut rose cultivrs over time. Flowers were hydrted in di wter overnight, re-cut to 6 cm in length nd plced inside glss mson jrs in vil with di wter. Jrs were closed for 2 hours t 21 o C. 2. Allure Lovely Drem Erin Red senstion Osin 15. 1. 5.. Pge 9 of 12
respirtion rte (ml/kg/h) respirtion rte (ml/kg/h) respirtion rte (ml/kg/h) Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 4 Respirtion rte of cut rose cultivrs fter ethylene exposure (1pmm) for 24 hours t 21 o C. Prior to externl ethylene exposure flowers were hydrted in di wter, 2.mM STS or 1 pmm AVG. Mens nd stndrd error n=3. 25. 2. Lovely Drem c2h4 di c2h4 sts c2h4 vg 25. 2. Erin c2h4 di c2h4 sts c2h4 vg 15. 15. 1. 5.. 1. 5.. 25. 2. 15. 1. 5. Red Senstion c2h4 di c2h4 sts c2h4 vg. Pge 1 of 12
Totl production/flower Totl production/flower Totl production/flower Totl production/ flower Totl production/flower Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 5. Totl voltile production over time of cut rose cultivrs over time. Flowers were plced in solution of di wter. Mens nd stndrd error of two flowers.. Decresed b. Incresed 4 35 3 25 2 15 1 5 Allure exp 16 2 4 6 ir-di ir-sts ir-vg 3 25 2 15 1 5 Lovely Drem 2 4 6 ir-di ir-sts ir-vg 4 35 3 25 2 15 1 5 Osin 2 4 6 ir-di ir-sts ir-vg 8 7 6 5 4 3 2 1 Erin 2 4 6 ir-di ir-sts ir-vg 35 3 Red Senstion ir-di ir-sts ir-vg 25 2 15 1 5 2 4 6 Pge 11 of 12
Totl production/flower Totl production/flower Totl production Totl production/flower Totl production/flower Totl production Ethylene, Frgrnce nd Vse Life of Cut Rose Flowers 9/27/216 Figure 6. Effects of inhibitors of ethylene synthesis (AVG) nd ethylene perception (STS) on totl voltile production over time of cut rose cultivrs fter ethylene exposure (1pmm) for 24 hours. Prior to externl ethylene exposure flowers were hydrted in di wter, 2.mM STS or 1 pmm AVG. Mens nd stndrd error n=2.. Effects of AVG on totl voltile production b. Effects of STS on totl voltile production 45 4 35 3 25 2 15 1 5 Allure exp 16 2 4 6 ehtylene-di ehtylene-vg 3 25 2 15 1 5 Lovely Drem 2 4 6 ehtylene-di ehtylene-sts 3 25 2 15 1 5 Lovely Drem 2 4 6 ehtylene-di ehtylene-vg 6 5 4 3 2 1 Erin 2 4 6 ehtylene-di ehtylene-sts 8 7 6 5 4 3 2 1 Erin 2 4 6 ehtylene-di ehtylene-vg 35 3 25 2 15 1 5 Red Senstion 2 4 6 ehtylene-di ehtylene-vg Pge 12 of 12