299-313 Issn 2077-4613

Middle East Journal of Applied Volume : 07 | Issue :02 |April-June| 2017 Sciences Pages: 299-313 ISSN 2077-4613

Influence of Nanosilver and Stevia extract on cut Anthurium Inflorescences

Ola A. Amin

Ornamental Plants and Landscape Gardening Res. Dept., Hort. Res. Inst., ARC, Egypt. Received: 23 March 2017 / Accepted: 28 April 2017 / Publication date: 30 April 2017

ABSTRACT

This is study was conducted in the Postharvest Lab of Ornamental Plants and Landscape Gardening Res. Dept., Hort. Res. Inst., Giza, Egypt in March of 2014 and 2015 seasons, aiming to assess the efficiency of some antiseptic solutions to improve the quality of Anthurium andraeanum inflorescence. Cut flowers were immersed for 3 h in benzyladenine (BA) at 10 mg l-1 plus sucrose at 25 g l-1 and then transferred to the following antiseptic solutions viz. nano-silver (NS) at 5, 10 and 15 mg l-1, -1 -1 calcium chloride (CaCl2) at 0.5, 1.0 and 1.5 g l + 8- hydroxyquinoline (HQ) at 0.2 g l and Stevia rebaudiana extract at 0.1, 0.2 and 0.3 mg l-1 all of these at different concentrations and combinations were combined with sucrose at 20 g l-1. Distilled water was used (D.W) as control. Data revealed that treatment of cut flowers with NS at 15 mg l-1 prolonged shelf life, maintained relative fresh weight and water balance, raised percentage of dry matter of flowers, reduced microbial count, enhanced some visual characteristics such as spadix condition as spathe blueing, spathe glossiness and stem condition. It had positive effects on total sugars and total carbohydrates content whilst Stevia extract at 0.1mg l-1 increased the rate of water uptake after fifteen days from placing the flowers in the solution. NS at 10 mgl-1 decreased water loss.

Key words: Anthurium andraeanum, nanosilver (NS), 8-hydroxyquinoline (8-HQ), plant extracts , benzyladenin (BA), calcium chloride (CaCl2).

Introduction

Anthurium cut flowers (Anthurium andraeanum L.) rank ninth in the global flower trade. The genus Anthuriums are evergreen plants belonging to the family Araceae. The plant possesses an underground rhizome with adventitious roots, characteristic of the family. Anthuriums characteristically produce numerous inflorescences, subtended by brightly colored spathes which are carried on long, slender peduncles. Spathes are characteristically heart-shaped, flat, puckered and shiny. Flowers have a wide range of spathe colors, viz. white, pink, salmon-pink, red, light-red, dark-red, brown, green, lavender, cream or multi-colored. The colorful spathe is long-lasting. However, the ‘true’ flowers are found on the spadix and have a large number of pistils (Islam et al., 2013).Vase life is therefore, determined by a complex criterion based on whichever of these symptoms appears first. Paull and Goo (1985) ascribed the symptoms of cut flower senescence in anthurium, that is loss of spathe glossiness, spathe and spadix browning and spathe wilting due to water stress. Regarding another symptom which is spathe blueing, it is attributed to an increase in pH caused by an increase in ammonium ions, resulting from the breakdown of proteins (Paull et al., 1985). He et al. (2006) indicated that vase life termination of cut flowers is characterized by wilting which is due to loss of water from the cells and low water uptake which is often due to occlusions located mainly in the basal stem end as affected by bacteria in vase water that may plug the vessels in the cut surface of stems (Ferrante et al., 2007). Van Doorn (1997) cleared that microbes are a common cause of stem end blockage, and as a result, water absorption and stem turgidity are reduced. It has been stated that microbial contamination, the cause of vascular occlusion, is the most limiting factor in longevity of cut flowers. Several natural and chemical substances have been applied to inhibit bacterial growth and so prolong the vase life of cut flowers. Floral preservative are mainly comprised of sugar and biocide source. Many agents have been used in vase solutions of the cut flowers to extend vase life by improving water uptake. Recently several natural and chemical substances have been applied to inhibit

Corresponding Author: Ola A. Amin, Ornamental Plants and Landscape Gardening Res. Dept., Hort. Res. Inst., ARC, Egypt. E-mail: [email protected] 299 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613 bacterial growth and enhance vase life. 8-hydroxyquinoline (8-HQ) and 8-hydroxyquinoline citrate (8-HQC) are commonly used as biocides. 8-HQ has been known to possess strong antimicrobial properties that eliminate vascular blockage and enhance water uptake so as to maintain water balance by reducing transpiration from flower tissue (Rogers 1973 and Jowker, 2005). The beneficial effect of sugars on flower senescence was attributed to the supply of substrates for respiration, structural and osmotic materials (Pun and Ichimura, 2003). Khandaker et al. (2017) showed that 6% sucrose was the best treatment for maintaining the post-harvest quality as well as vase life of cut Mokara Chark Kuan orchid flowers. Therefore, it is important to use certain materials in vase solutions to extend the vase life of cut flowers as Ca may increase longevity of cut flowers by delaying physiological events related to senescence, such as decreasing water uptake, increased water transpiration loss, decreased fresh weight, stem bending, or prevention of disease during propagation. Using calcium in vase solutions increases water flow through the stems by association with pectin in the xylem cell walls and delay the degradation processes that affect cell membrane integrity (Grignon and Sentenac, 1991; and Van Ieperen and Van Gelder, 2006). Sairam et al. (2011) reported that the vase life of gladiolus flowers was increased by calcium chloride holding solution. Using nano materials including nanosilver has recently increased in the world. Use of NS is becoming increasingly widespread in medicine and various other industrial purposes. Usage of nano-silver compounds (NS) in pulse and vase solution treatment for cut flowers is relatively new (Liu et al., 2009 and Solgi et al., 2009) and has demonstrated its importance as an antibacterial agent (Alt et al., 2004 and Morones et al., 2005). It has been widely used due to its anti bacterial property. It also has the additional benefit of high durability, more simple and easy to use than other anti-bacterial agents (Van Doorn, 1997). Generally Świder et al. (2017) declared that the preservatives delayed reduction in fresh weight. Bahrehmand et al. (2014) suggested that NS has the following effects; reduces transpiration in association with reducing stomata aperture (stomata closure), increases hydraulic conductance, inhibits bacterial growth in the vase solution and at the cut stem end, prevents ethylene-mediated processes, such as flower senescence. These effects of NS are due to its particle size, with higher surface to volume ratio. Mortazavi et al.(2011) and Moradi et al. (2012) found that nano-silver in combination with sucrose increase the vase life of cut flowers. Natural plant extracts are safe, cheap and are suitable alternatives for conventional substances used to prolong vase life of cut flowers. Using plant extracts such as the extract of Stevia rebaudiana leaves, may have a role to be used as pharmaceutical and or preservatives. Stevia rebaudiana Bertoni, a natural sweetener plant with zero calorie content, becomes an inevitable alternative to sugar especially with the over 346 million diabetic population across the world (Abdullatef and Osman, 2012). Stevia has been reported to have no adverse effect on human (Brandle and Rosa, 1992). The leaves could be eaten fresh or when dried and it could be boiled in tea to release the sweetener. Godarzi (2006) reported that the extract obtained from dried leaves of stevia in acetone has stronger antibacterial properties than it's aqueous extract. BA effectively delayed leaf yellowing and also tepal senescence in tulips (Van Doorn et al., 2011). Fukui et al. (2005) indicated that BA treatment is mostly effective in extending vas life of cut anthurium. Nouri et al. (2012) cleared that benzyladenine prevent ethylene production and it is the most important factor in delaying senescence. The present study was conducted with an objective to prolong the postharvest longevity of anthurium cut flowers, maintaining the flower quality and reducing bacteria using holding solution in association with antibacterial agent.

Materials and Methods

The present trial was undertaken at the Postharvest lab of Ornamental Plants and Landscape Gardening Res. Dept., Hort. Res. Inst., Giza, Egypt in March of 2014 and 2015 seasons to find out the response of anthurium cut flowers to some preservative solutions.

Plant material: Cut flower stems of anthurium Anthurium andraeanum L. cv. Fire (red spathe) was freshly obtained from the local commercial green houses of Floramix Farm (El-Mansouria, Giza) in each season. Cut flowers were picked in the early morning in 40% open stage and were directly wrapped in groups inside carton boxes (102 x 50 x 30 cm) and transported quickly to the laboratory within nearly 2h. As soon as in the lab,

300 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613 cut stems were precooled by placing in cool water for half an hour to remove the effect of high field heat. Thereafter, stem bases were recut under water to avoid air embolism before treatments, and flowers were adjusted to the same size (stem length of 65cm), shape. Only injury free stems were selected for the experiment.

Experiment treatments:

Stems of flowers were pulsed for three hours in benzyl adenine (BA) at 10 mg/l plus sucrose at 25 g/l and symbolize it with A1 and then, flowers were placed in a glass bottle (500 ml) containing 400 ml of the following antiseptic solutions as holding with the different levels viz.: 1- Distilled water (control). 2- Nano-silver (NS) at 5 mg/l plus sucrose (suc) at 20 g/l. 3- Nano-silver (NS) at10 mg/l plus sucrose (suc) at 20 g/l. 4- Nano-silver (NS) at15 mg/l plus sucrose (suc) at 20 g/l. 5- Calcium chloride (CaCl2) at 0.5 g/l + 8- hydroxyquinoline (HQ) at 0.2 g/l + sucrose (suc) at 20 g/l. 6- Calcium chloride (CaCl2) at1g/l + 8- hydroxyquinoline (HQ) at 0.2g/l + sucrose (suc) at 20g/l. 7- Calcium chloride (CaCl2) at1.5g/l + 8- hydroxyquinoline (HQ) at 0.2g/l + sucrose (suc) at 20g/l. 8- Leaf extract of Stevia rebaudiana at 0.1 mg/l with sucrose (suc) at 20 g/l. 9- Leaf extract of Stevia rebaudiana at 0.2 mg/l with sucrose (suc) at 20 g/l. 10- Leaf extract of Stevia rebaudiana at 0.3 mg/l with sucrose (suc) at 20 g/l. After that, each bottle was covered at its mouth with cellophane wrap to prevent evaporation. Flowers were arranged in a completely randomized design of 10 treatments and 3 replications.

Experiment measurements :

 Shelf life: The time from the start of treatment until the senescence of flowers (days).  Water loss: Cumulative water loss was recorded for the entire period of vase life of the flower stalk (g/flower).  Water uptake: Cumulative water uptake was recorded for the entire period of vase life of the flower stalk (g/flower).  Water balance: Water balance (g/flower) =water uptake - water loss.  Relative fresh weight: The fresh weight of cut flowers was recorded daily during the experiment. Relative fresh weight of stems was calculated using the following formula (He et al., 2006):

RFW (%) = Fresh weight of stem in mentioned day ×100 Fresh weight of stem in day zero

 Dry matter: At the end of flower vase life, fresh weight of flower of each treatment was calculated and then dried to a constant weight in an oven for 24 hr at 72°C. Dry matter percentage of cut flowers was calculated by the following equation: DM (%) = dry weight/fresh weight × 100 (Hashemabadi et al., 2015).  Postharvest evaluation: Visual assessment of quality was applied according to procedures used by Paull (1982). This appraisal consists of scoring anthurium for loss of spathe gloss, spathe blueing and spadix necrosis as listed in the following tables ( A,B,C and D).

Table A: Grading scale for spadix visual condition. Score Spadix condition 1 No spots 2 Spadix tip showing slight discoloration/darkening 3 Spadix tip showing darkening/slight flower separation 4 Less than 10% of spadix length darkened and dry 5 Spadix tip dryed and necrosed in more than 10% of its lenght

301 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

Table B: Grading scale for spathe blueing . Score Spathe condition 1 None – fresh flower looks, no blueing spadix tip showing Slight – less than 5% blueing 2 area spadix tip showin 3 Moderate – 5 to 10 % blueing area 4 Severe – more than 5% blueing area

Table C: Grading scale for spathe glossiness. Score Spathe condition 1 No loss – high gloss stage right after harvest 2 Slightly loss – not perceivable 3 Moderate loss – slight glossiness remain 4 Severe loss – flat spathe, no glossiness and wilting signs may appear

Table D: Grading scale for stem condition. Score Stem condition 1 Good fresh stem 2 Slightly browning and bending 3 Moderate browning and bending 4 Severe browning 5 All dried

Postharvest evaluation was determined by numerating the days before a stem reached grade 5 for spadix condition and grade 3 of spathe blueing and grade 3 for spathe glossiness and grade 3 for stem condition.  Microbial count: Microbial count was determined by taking 1ml vase solution samples during the first 6 days of the experiment, at 2 days intervals with 3 replications. One ml from each sample was diluted in 10 fold serial dilution. 0.1 ml from each concentration of diluted samples was plated on nutrient agar and all were incubated at 35 ºC for 48 hours. Microorganisms were counted by standard plate counting method (by counting the number of colonies formed after incubation) to generate the number of colony forming units.ml–1 (CFU ml–1) according to Jowkar (2006).  Chemical analyses, total soluble sugars (%) and total carbohydrate (mg/g) were evaluated using the method of Dubois et al. (1956). Total phenols (mg/g) were determined according to Singleton et al. (1999).  To prepare leaf extract of stevia, 150 g dry leaves of stevia were poured in 500 ml acetone plus 500 ml distilled-water and was settled on vibration machine (Labcon model) for 72 hours. The mixture was then filtered by a filtration paper. The filtrate was put in the oven at 70 ºC temperature for 72 hours to dry and change to a uniform powder (Gosh et al., 2008).

Experiment condition: ¹־²S־Flowers were placed in ambient conditions 22±1 ºC, light level was about 20 µ mol m partially from natural light and partially from fluorescent cool light for 12h/day.

Statistical analysis: Data were tabulated and statistically analyzed according to SAS Institute program (2009) followed by Duncan`s New Multiple Range Test (Steel and Torrie, 1980) to verify the significance among means of different treatments.

Results and Discussion

Shelf life

Results showed that the application of different antiseptic preservative solutions was effective in extending vase life period of cut anthurium flowers in comparison to the control as documented in

302 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

Table (1). The best result in this regard was obtained from the treatment of 15 mg l-1NS combined with -1 -1 -1 sucrose at 20 g l +A1 (pulse flowers for three hour in benzyl adenine at 10 mg l plus sucrose at 25 g l ) that gave 24 days in the first season and 23.67 days in the second one as it seems to control senescence, whilst the control treatment gave 18 days in the first season and 18.67 days in the second one. A similar effect was a resulting of using NS at 10 mg l-1plus sucrose at 20 g l-1 which enhanced shelf life of the cut flowers compared to the control. This may be to the positive effect of NS treatments to inhibit bacterial growth in the vase solution and at the cut surface stems during shelf life period. The fresh weight of cut flowers diminished as they approach the aging step. This may be related to the reduction of water absorption and the loss through petals and leaves turgidity. The current findings support this idea, Hashemabadi (2014) and Basiri et al. (2011) showed that the silver nano-particles treatment inhibited bacterial growth in the preservative solution and enhanced vase life of cut carnation. Many authors indicated the role of NS in prolonging shelf life of cut flowers as confirmed by Liu et al. (2009) and Solgi et al. (2009) on gerbera cultivars ; Lü et al. (2010) and Hatami et al.(2013) on rose and Roshani et al. (2016) on cut carnation.

Table 1: Effect of antiseptic solution treatments on shelf life of Anthurium andraeanum flowers (days) during the vase life period of 2014 and 2015 seasons. Vase life (days) Treatment 1st Season 2nd Season

Distilled water (control) 18.00D 18.67E

NS (5mg l-1) + suc (20g l-1) 21.67B 22.33B NS (10mg l-1) + suc (20g l-1) 23.33A 23.33A NS (15 mg l-1) + suc (20g l-1) 24.00A 23.67A

-1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 20.67C 20.00D

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 22.00B 22.33B

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 22.00B 21.67BC Stevia extract (0.1mg l-1 )+ suc (0g l-1) 20.67C 21.67BC Stevia extract (0.2mg l-1 )+ suc ( 20g l-1) 20.00C 20.00D Stevia extract (0.3mg l-1)+ suc (20g l-1) 17.33D 17.33F Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

Water loss

According to data presented in Table (2), flowers treated with stevia extract at 0.1 mg l-1+ suc at 20 -1 -1 g l +A1 had lower rate of water loss after 1 day of holding in vase solution and gave 2.65 - 2.75 g l compared to 3.8 - 4.14 g l-1 in the control treatment with significant differences in the two seasons. On the other hand, data in the same table indicated that NS at 5 mg l-1 + suc at 20g l-1 was the most effective on decreasing water loss of cut flowers compared with other holding solutions throughout the seven days after holding stems in vase solution, in both seasons. It was also observed that 15 days after treating flowers with nano silver and calcium chloride at the different concentrations reduced water depletion from cut flowers especially NS at 5 and10 mg l-1 + suc -1 at 20 g l + A1 that produced the best results, compared to the control. This may imply that NS treatments decreased stomatal conductance and transpiration rate of the cut rose probably due to stomatal closure induced by NS. The aforementioned results are in well agreement with Lü et al. (2010) on cut rose. Likewise, Rafi and Ramezanian (2013) declared that applying NS pulse treatments reduced water loss, stomatal conductance and transpiration rate in Rosa. hybrida L. cvs. ‘Avalanche’ and ‘Fiesta’.

303 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

Table 2: Effect of antiseptic solution treatments on water loss of Anthurium andraeanum flowers (g/flower) during the vase life period of 2014 and 2015 seasons. Water loss

1st Season 2nd Season Treatment After1day After7days After15days After1day After7days After15days

Distilled water (control) 3.850A-C 16.48B 25.22C 4.147CD 16.50CD 25.41D NS (5mg l-1) + suc (20g l-1) 4.397A 15.27CD 24.24D 5.670A 13.78E 21.60G NS (10mg l-1) + suc (20g l-1) 3.480BC 13.73F 22.11E 3.810DE 14.20E 22.44F NS (15 mg l-1) + suc (20g l-1) 3.797A-C 14.56DE 25.23C 3.718DE 15.78D 25.40D

-1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 4.190AB 16.42B 24.15D 4.237CD 17.42AB 25.28D

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 3.983AB 14.07EF 22.12E 5.203AB 16.52CD 25.44D

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 4.270A 14.60DE 22.61E 4.683BC 15.96D 24.32E Stevia extract (0.1mg l-1)+ suc (20g l-1) 2.650D 16.04BC 27.68B 2.750F 16.49CD 28.46B Stevia extract (0.2mg l-1)+ suc (20g l-1) 2.720D 16.58B 27.96B 3.213EF 18.09A 29.89A Stevia extract (0.3mg l-1)+ suc (20g l-1) 3.143CD 18.49A 29.56A 3.24EF 16.89BC 27.63C Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

Water uptake

As shown in Table (3) data indicated that cut stems held in antiseptic solution containing NS at 5 -1 -1 mg l plus sucrose at 20 g l after pulsing in A1 absorbed the highest amount of water and gave 4.580 g/stem in the first season and 5.793 g/stem in the second one compared to 3.897 and 4.227 g/stem from the control, respectively in the two seasons, after one day of holding it in the solutions. This may be reasonable because of the role NS which may positively influence on water uptake, besides the anti-bacterial effect. Similar findings were revealed by Shah and Belozerova (2009) who found that large surface to volume ratio of NS particles increase their contact with fungi or bacteria, and vastly improving their antifungal and antibacterial efficiencies.

Table 3: Effect of antiseptic solution treatments on water uptake of Anthurium andraeanum flowers (g/flower) during the vase life period of 2014 and 2015 seasons. Water uptake Treatment st nd 1 Season 2 Season After1day After7days After15days After1day After7days After15days

Distilled water (control) 3.897BC 12.10F 18.97E 4.227CD 14.27D 21.72D NS (5mg l-1) + suc (20g l-1) 4.580A 15.85CD 24.64C 5.793A 14.35D 22.20D NS (10mg l-1) + suc (20g l-1) 3.607CD 14.30E 22.57D 3.927CD 14.73D 22.97D NS (15 mg l-1) + suc (20g l-1) 3.823BC 15.03D 25.76B 3.817CD 16.41C 26.17B

-1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 4.370AB 17.11B 24.45C 4.373BC 17.92AB 25.42BC

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 4.170A-C 14.66DE 22.07D 5.373AB 17.25BC 25.59BC

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 4.500A 15.25D 22.61D 4.85A-C 16.62C 24.45C Stevia extract (0.1mg l-1)+ suc (20g l-1) 2.690E 16.37C 27.76A 2.763E 16.77C 28.55A Stevia extract (0.2mg l-1)+ suc (20g l-1) 2.793E 17.05B 26.00B 3.260DE 18.51A 28.84A Stevia extract (0.3mg l-1)+ suc (20g l-1) 2.67E 18.81A 27.60A 3.313DE 17.16BC 25.97B Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

Hatami et al. (2013) reported that silver nano particles may have positive influence on water uptake because of antibacterial effects of Ag+ ions that as may affect the regulation of water channel activity via inhibition of sulfhydryl-containing proteins and improve solution uptake. Previous results are confirm with those attained by Bahrehmand et al. (2014) who mentioned that flower relative water

304 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613 content increased under sucrose application, and tuberose flower water uptake increased by NS application. Vinodh et al. (2013) and Nemati et al. (2014) postulated that nano silver induced the highest amount of water uptake of Lilium orientalis. On the other side, data of second period (after7 days) revealed all treatments raised the amount of water uptaken by cut stems with various differences as compared to the amount uptaken by control cut stems. The superiority was to stevia extract at 0.3 mg l-1+ suc at 20 g -1 -1 l +A1 in the first season. This excess was a result of treating cut flowers by stevia extract at 0.2 mg L + -1 suc at 20 g l +A1 in the second season. In relation to the third period of shelf life of cut anthuruim, stevia extract at various concentrations enhanced the amount of water uptake. This may be attributed to the antibacterial compounds in stevia extract, which inhibited vascular blockage and increased absorption process in flowers. Eshaghdavatgar et al. (2013) cited that stevia extract inhibited vascular blockage in the vase solution of cut rose flowers cv. "Dolce Vita".

Relative fresh weight

From data averaged in Table (4) it is clear that in the first period (one day after putting flowers in the antiseptic solutions) the most effective antiseptic solutions in maintaining relative fresh weight of -1 -1 -1 cut flowers were the combination of CaCl2 at 1.5 g l + HQ at 0.2 g l + suc at 20 g l + A1 followed by -1 -1 NS at 5 mg l + suc at 20 g l + A1 with significant difference compared to the control, as they improved the relative fresh weight. This may be due to the role of calcium which decreased the activity and effect of ethylene on cell walls and inhibit cell membrane injury and ethylene synthesis, leading to improve the strength of plant cell wall (Mortazavi et al., 2016).

Table 4: Effect of antiseptic solution treatments on relative fresh weight of Anthurium andraeanum flowers (%) during the vase life period of 2014 and 2015 seasons. Relative fresh weight 1st Season 2nd Season Treatment After1day After7days After15days After1day After7days After15days

Distilled water (control) 100.28C 100.68E 94.04G 100.43A-C 101.03F 97.45D NS (5mg l-1) + suc (20g l-1) 100.96A 102.90A 102.01B 100.82A 103.38A 102.73B NS (10mg l-1) + suc (20g l-1) 100.93A 102.80AB 101.93B 100.53AB 102.53C 102.43B NS (15 mg l-1) + suc (20g l-1) 100.12C 102.42C 102.78A 100.52AB 102.96B 103.66A

-1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 100.68AB 102.55A-C 97.69F 100.63A 102.24CD 96.81E

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 100.79A 102.50BC 99.57E 100.73A 103.14AB 100.63C

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 101.01A 102.83AB 100.02D 100.67A 102.55C 100.54C Stevia extract (0.1mg l-1)+ suc (20g l-1) 100.20C 102.03D 100.64C 100.09C 101.82E 100.58C Stevia extract (0.2mg l-1)+ suc (20g l-1) 100.37BC 102.21CD 89.74H 100.22BC 101.94DE 94.22F Stevia extract (0.3mg l-1)+ suc (20g l-1) 100.34BC 102.00D 87.76I 100.50AB 101.88E 89.72G Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

These results agreed with those of Cortes et al. (2011) who showed that using CaCl2 in the vase water increased the fresh weight of cut rose. Ibrahim et al. (2011) who concluded that the chemical solutions of 8-HQS at 100 or 200 ppm or CaCl2 at 1000 or 2000 ppm, each supplemented with 4% sucrose lowered the flower weight loss. In regard to the second period (seven days of the presence of flowers in solutions) all treatments caused a positive increment in relative fresh weight percentage. The highest increase in both seasons was gained by holding flowers with NS at 5mg L-1 combined with suc (20g l-1). On the other side, the lowest percentage was obtained by using stevia extract (0.3 mg l-1) + suc (20 g l-1) -1 +A1 recording 87.76 and 89.72% in the first and second seasons. NS at 15 mg l combined with suc at 20g l-1 gave higher rate compared to the control and the other treatments during the 15 days period of the existence of flowers in the solution. Typically, cut flowers initially increase and subsequently decrease in RFW. This is probably attributed to lack of food in the solution, thus forcing the flower to use all its reserves. It also could be due to the decrease in water uptake (Alaey et al., 2011 and Thwala et al., 2013)

305 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

SNP may have a positive influence on water uptake because of the antibacterial effects of Ag+ ions in SNP, thereby inducing higher percentage of fresh weight of cut anthurum flowers. Several reports are also in accordance with the previous gains, such as those of Hatami et al.,(2013) and Amingad et al. (2017) who stated that SNP gave highest relative fresh weight (%) in cut rose. Similar reports were declared by Nemati et al. (2014) on Lilium orientalis.

Water balance

It is evident from data presented in Table (5) that treatment with calcium chloride at 1.5 g l-1 in -1 -1 addition to HQ at 0.2g l , suc at 20g l and A1 achieved the highest values during the first vase life period, as it gave 0.23- 0.17 g l-1 compared to 0.04- 0.08 g l-1 resulting from the control treatment, respectively in two seasons of this investigation. A similar trend was also obtained regarding the effect of calcium chloride at various concentrations accomplished in the second period (seven days from the beginning). Increasing the rate of water balance in presence of antiseptic solutions may be attributed to the role of HQ as described by Jowkar et al. (2012) who found that HQC is known to inhibit ethylene production in cut flowers and suppressed bacterial growth in vase solution. The role of CaCl2 was cleared by Hassani and Alimirzaii (2017) who observed that using calcium in vase solutions increases water flow through the stems by association with pectin in the xylem cell walls of cut rose. Therefore, an improvement of its absorption condition leads to positive state of water equilibrium in cut flowers. This finding was demonstrated before by Sven and Jose (2004) who stated that sucrose helps to maintain the water balance. Soumen and Roychowdhury (2005) noted that the best water balance was observed with the flowers of Anthurium andraeanum treated with silver nitrate 50 ppm + 8-HQ 400 ppm + sucrose 5%. Hassani and Alimirzaii (2017) who observed that using calcium in vase solutions increases water flow through stems by association with pectin in the xylem cell walls. In the last stage of shelf life of anthurium, it was observed that NS stimulated water condition and -1 -1 maintained water balance of cut anthurium flower was affected by NS (15 mg l ) + suc (20g l ) +A1. These findings agreed with the findings of Lü et al. (2010) who noticed that pulse treatment in 10 mg l-1 NS plus 5% sucrose solution for 24 h followed by holding in 0.5 mg l-1 NS plus 2% sucrose solution not only alleviated and delayed vascular blockage caused by bacterial contaminations, but also inhibited stomatal conductance so that the water balance in cut roses was significantly improved.

Table 5: Effect of antiseptic solution treatments on water balance of Anthurium andraeanum flowers (g/flower) during the vase life period of 2014 and 2015 seasons. Water balance Treatment 1st Season 2nd Season

After1day After7days After15days After1day After7days After15days Distilled water (control) 0.0466C -4.380E -6.250F 0.080B-D -2.230G -3.690G NS (5mg l-1) + suc (20g l-1) 0.183AB 0.5800B 0.4033AB 0.1233A-C 0.5767CD 0.6033B NS (10mg l-1) + suc (20g l-1) 0.127A-C 0.5700B 0.4700A 0.1167A-C 0.5267D 0.5333B NS (15 mg l-1) + suc (20g l-1) 0.0266C 0.4667C 0.5300A 0.1000B-C 0.6300BC 0.7733A

-1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 0.180AB 0.6900A 0.3033B 0.1367AB 0.5067D 0.2233C

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 0.186AB 0.5900B -0.056D 0.1700A 0.733A 0.1500CD

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 0.2300A 0.653AB 0.006CD 0.1700A 0.6567B 0.1333CD Stevia extract (0.1mg l-1)+ suc (20g l-1) 0.0333C 0.330D 0.1000C 0.0133E 0.2833F 0.0900D Stevia extract (0.2mg l-1)+ suc (20g l-1) 0.073BC 0.4733C -1.963E 0.0466DE 0.42E -1.050E Stevia extract (0.3mg l-1)+ suc (20g l-1) -.476D 0.3200D -1.960E 0.0733C-D 0.2733F -1.657F Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

Dry matter

Data represented in Table (6) exhibited that anthurium cut flowers held in NS at 15 mg l-1 + suc at -1 20 g l +A1 had significantly the highest values in both seasons. It is the best treatment in increasing dry matter percentage. Cut flowers held in distilled water (control) had the lowest dry weight content. This

306 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613 may be due to the role of sucrose in providing the required energy for survival of flowers and affecting the structure of the flower cell walls and delays aging through this and causing an increase in the dry weight and water retention. Compounds extending the vase life of cut flowers on the prevention of dry weight loss by preventing the degradation of carbohydrates (Dashtbany and Hashemabadi,2015).

Table 6: Effect of antiseptic solution treatments on dry matter of Anthurium andraeanum flowers (%) during the vase life period of 2014 and 2015 seasons. Dry matter (%) Treatment 1st Season 2nd Season

Distilled water ( control) 26.42G 26.97E NS (5mg L-1) + suc (20g l-1) 34.80C 35.04B NS (10mg L-1) + suc (20g l-1) 38.52B 34.23B NS (15 mg L-1) + suc (20g l-1) 40.48A 39.57A -1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 30.46E 30.71C

-1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g L )+ suc (20g l ) 29.27EF 27.07E

-1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc (20g l ) 31.89D 31.50C Stevia extract (0.1mg l-1)+ suc (20g l-1) 28.32F 28.37D Stevia extract (0.2mg l-1)+ suc (20g l-1) 28.6F 27.39DE Stevia extract (0.3mg l-1)+ suc (20g l-1) 28.27F 27.67DE Means followed by the same letter/s in a column or raw do not differ significantly according to Duncan's New Multiple Range test at P =0.05

Microbial count

As shown in Table (7) data indicated that in regard to the population of bacteria in vase solution, the presences of NS reduced bacterial growth in vase solution and there was a positive relationship between the number of bacteria in the preservative solution and vase life of cut flowers. Treating -1 -1 flowers with NS (15 mg l ) combined with suc (20g l ) and A1 gave the best result and inhibited microorganisms in vase solution to the lowest level.

Table 7: Effect of antiseptic solution treatments on microbial count of Anthurium andraeanum flowers (%) during the vase life period of 2014 and 2015 seasons. Treatments Result Distilled water (control) 8.0 x106 NS (5mg/l) + suc (20g/l) 1.7x102 NS (10mg/l) + suc (20g/l) 1.5x 102 NS (15 mg/l) + suc (20g/l) 1.3x 102 2 CaCl2(0.5g/l)+ HQ (0.2g/l)+ suc (20g/l) 5.0x 10 2 CaCl2(1.0g/l)+ HQ (0.2g/l)+ suc (20g/l) 4.8x10 2 CaCl2(1.5g/l)+ HQ (0.2g/l)+ suc (20g/l) 4.2x 10 Stevia extract (0.1mg/l)+ suc (20g/l) 1.1x106 Stevia extract (0.2mg/l)+ suc (20g/l) 2.2 x106 Stevia extract (0.3mg/l)+ suc (20g/l) 3.7 x106

On the other side, the maximum average of bacterial count was recorded by control treatment. In accordance with the previous gains, Furno et al. (2004) affirmed that silver nanoparticle (SNP) as a pulse treatment for cut flowers has demonstrated importance as an anti bactericidal agent that could kill 650 species of bacteria in water. Rai et al. (2009) reported that silver nano-particles (SNPs) show efficient antimicrobial property compared to other salts due to their extremely large surface area, which provides better contact with microorganisms. Oraee et al. (2011) reported that nano-silver can be used to decrease bacteria in stem end of cut gerbera and in solution. Similar results were obtained by Kazemi and Ameri (2012) on cut rose; Hatami et al. (2013) on cut carnation, Nemati et al. (2013) on Lilium orientalis. Li et al. (2017) found that the 25 mg l−1 NS pulse treatment effectively inhibited bacterial colonization and biofilm formation on the stem-end cut surface and in the xylem vessels of cut gladiolus spikes.

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Postharvest evaluation:

Treatment 3-nano-silver (NS) at15 mg/l plus sucrose (suc) at 20 g/l +A1 followed byT3-nano silver (NS) at10 mg/l plus sucrose (suc) at 20 g/l +A1 achieved the longest period until the point of non-acceptance of flowers (Fig,1). There was also an increase in the marketable flowers at 15 mg/l and in parallel with NS at 10 mg/l, compared to control, as flowers were considered marketable if they showed less than 10% spathe blueing (Fig,2). Spathe glossiness and stem condition were enhanced by treading flowers with NS at 15 mg/l obviously compared to the control (Fig 3,4). Anthurium quality can be defined as vase life, as well as physical attributes like spadix visual condition, spathe blueing, spathe glossiness and stem condition. In confirmation to the current experiment Elibox and Umaharan (2008) demonstrated that spadix necrosis, and spathe blueing/necrosis were the only signs of senescence common to 26 anthurium cultivars studied. Hettiarachchi and Balas (2005) investigated anthurium flowers and found that flower colour (both spathe and spadix) was positively correlated with use of vase solutions.

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.

Distilled water (control). T1-Nano-Silver (NS) at 5mg/l plus sucrose (suc) at 20g/l. T2Nano-Silver (NS) at10mg/l plus sucrose (suc) at 20g/l.

T3-Nano-Silver (NS) at15mg/l plus sucrose (suc) at 20g/l. T4-Calcium chloride (CaCl2) at 0.5g/l + 8- hydroxyquinoline (HQ) at 0.2g/l + sucrose (suc) at 20g/l. T5-Calcium chloride (CaCl2) at1g/l + 8- hydroxyquinoline (HQ) at 0.2g/l + sucrose (suc) at 20g/l. T6-Calcium chloride (CaCl2) at1.5g/l + 8- hydroxyquinoline (HQ) at 0.2g/l + sucrose (suc) at 20g/l. T7Extract leaves of Stevia rebaudiana at 0.1mg/l with sucrose (suc) at 20g/l. T8-Extract leaves of Stevia rebaudiana at 0.2mg/l with sucrose (suc) at 20g/l. T9-Extract leaves of Stevia rebaudiana at 0.3mg/l with sucrose (suc) at 20g/l.

Chemical composition

Data in Table (8) illustrated that the most successful treatment was nano silver at 15 mg/l as it achieved the maximum values of carbohydrate content in the spadix and spathe (34.63-29.96 mg/g). On the other side, placing flower in distilled water gave the lowest values in this trait (13.14-12.56 mg/g). This result was consistent with the observation of Van Doorn (2004) who found that petal carbohydrate contents are one of the most crucial factors influencing the postharvest life of cut flowers. It is proposed that there is a positive correlation between the levels of endogenous sugars and the time to petal wilting. In addition, the senescence process of cut flowers is regulated by phytohormones and correlated with the carbohydrate status of the petals. Total sugars percentage in the spadix and spathe of anthurium cut flowers registered high significant increment as a result of treating with NS (15 mg/l) + suc (20 g/l) + A1 treatments compared to the control. These results are in agreement with those obtained by Bhanusree et al. (2015) who suggested that dipping cut gerbera in 5 % sucrose solution might be efficient in increasing total sugars content and consequently vase-life. Elgimabi (2014) mentioned that the combination of sucrose 7% + AgNO3 30% was significantly superior to the rest of combinations in retarding the chlorophyll as well as the carbohydrate degradation of Rosa damascena. Prashant et al. (2010) cited that the translocated sugars accumulated in the flowers increase the osmotic concentration and improve the sugar (total and reducing sugars) concentration and maintain turgidity.

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Stevia extract (0.3 mg /l) + suc (20 g/l) induced the highest value of phenolic content compared to control and other treatments.

Table 8: Effect of antiseptic solution treatments on phenols (mg/g), total sugars (%) and total charbohydrate (mg/g) of Anthurium andraeanum flowers during the vase life period of 2014 and 2015 seasons. Treatments Phenols (mg/g) Total sugars (% ) Total char.(mg/g) Spadix Spathe Spadix Spathe Spadix Spathe Distilled water (control) 0.169 0.147 15.70 13.92 13.14 12.56 NS (5mg l-1) + suc (20g l-1) 0.151 0.149 19.95 16.38 29.96 21.92 NS (10mgl-1) + suc (20g l-1) 0.139 0.127 20.64 18.23 34.15 29.10 NS (15 mg l-1) + suc (20g l-1) 0.124 0.138 21.48 19.78 34.63 29.96 -1 -1 -1 CaCl2(0.5g l )+ HQ (0.2g l )+ suc (20g l ) 0.166 0.155 16.69 15.63 29.02 16.54 -1 -1 -1 CaCl2(1.0g l )+ HQ (0.2g l )+ suc (20g l ) 0.157 0.145 17.23 17.62 31.77 19.53 -1 -1 -1 CaCl2(1.5g l )+ HQ (0.2g l )+ suc 20g l ) 0.125 0.114 18.06 18.97 32.88 25.73 Stevia extract (0.1mg l-1 )+ suc (20g l-1) 0.167 0.148 17.07 14.27 24.47 16.05 Stevia extract (0.2mg l-1 )+ suc (20g l-1) 0.163 0.126 15.57 16.83 23.24 14.72 Stevia extract (0.3mg l-1)+ suc (20g l-1) 0.180 0.164 12.30 11.76 13.63 11.54

Conclusion

-1 -1 According to the results obtained, it can be concluded that NS (15 mg l ) + suc (20 g l ) +A1 extended shelf life and delayed senescence as compared to other treatments. It maintained the performance of flowers according to various measurements taken as relative fresh weight, water balance, raised percentage of dry matter of flowers, reduced microbial count and improved postharvest evaluation of cut anthurium flowers. Hence, it could be recommended to hold cut flowering stems in -1 -1 nano silver at15 mg l + sucrose at 20 g l +A1 to obtain the best quality.

References

Abdullateef, R.A. and M. Osman, 2012. Studies on effects of pruning on vegetative traits in Stevia rebaudiana Bertoni (Compositae). Intl. J. Biol., 4(1): 146-153. Alaey, M., M. Babalar, R. Naderi and M. Kafi, 2011. Effect of pre and postharvest salicylic acid treatment on physio-chemical attributes in relation to vase life of rose cut flowers. Postharvest Biol. Tech., 61:91-94. Alt, V., T. Becher, P. Steinrucke, M. Wagener, P. Seidel, E. Dingeldein, E. Domann and R. Schnettler, 2004. An in vitro assessment of the antibacterial properties and cytotoxicity of nanoparticulate silver bone cement. Biomaterials., 25:4383-4391. Amingad, V., K.N. Sreenivas and K.S. Shivashankara, 2017. Comparison effects of silver nanoparticles, glycerol and aluminium sulphate as vase holding solutions on vase life attributes of cut rose (Rosa x hybrida) var. Taj Mahal. Intl. J. Agri. Sci. and Res., 7(2): 87-94. Bahrehmand, S., J. Razmjoo and H. Farahmand, 2014. Effects of nano-silver and sucrose applications on cut flower longevity and quality of tuberose (Polianthus tuberosa). Intl. J. Hort. Sci. and Tech., 1(1):67-77. Basiri, Y., H. Zarei and K. Mashayekhi, 2011. Effects of nano-silver treatment on vase life of cut flowers of carnation. J. Adv. Lab. Res. Bio., 1: 50-55. Bhanusree, M.R., N.H. Rao, M. Chandrika, M. Vinayakumari, K. R. Kumar, G. Shukla and S. Chakravarty, 2015. Effect of sucrose on biochemical parameters of cut Gerbera flowers (Gerbera jamesonii Bolus ex. Hook.) cv. Lamborgini J. Agric. Tech., 2(1&2): 68-71. Brandle, J.E. and N. Rosa, 1992. Heritability for yield, leaf-stem ratio and stevioside content estimated from a landrace cultivar of Stevia rebaudiana. Can. J. Plant Sci., 72(4):1263-1266. Cortes, M.H., A.A. Frias, S.G. Moreno, M.M. Pina, G.H.D.C. Guzman and S.G. Sandoval, 2011. The effects of calcium on postharvest water status and vase life of Rosa hybrida cv. Grand Gala - Int. J. Ag. & Biol.,13: 233-238. Dashtbany, S. and D. Hashemabadi, 2015. Study on interaction effects of mechanical and Geranium essential oil treatments on vase life of cut Chrysanthemum (Dendranthema grandiflorum L.). J. Orn.Pla., 5 (2): 97-103.

310 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

Dubois, M., F. Smith, K.A. Illes, J. K. Hamilton and P. A. Rebers,1956. Colorimetric method for determination of sugars and related substances. Annal. Chem., 28 (3):350-356. Elgimabi, M.N.E.E., 2014. Pulsing with sucrose and silver nitrate enhance water uptake and result in along vase life in Taif rose cut flowers (Rosa damascena cv. Trigintipetala). Intl. J.Agric. Sci., 6(1):379-383. Elibox, W. and P. Umaharan, 2008. Morphophysiological characteristics associated with vase life of cut flowers of anthurium. HortScience, 43(3):825–831. Eshaghdavatgar, L., M. Jafarpour and A.R. Golparvar, 2013. Comparing medicinal plant extracts and pulsing treatments on rose cut flowers "Dolce Vita". Sci. Agri., 4 (1): 1-4. Ferrante, A., A. Aberici, S. Antonacci and G. Serra, 2007. Effect of promoters and inhibitors of phenylalanine ammonia lyase enzyme on stem bending of cut gerbera flowers. Acta Hort., 755: 471-476. Fukui, R., S. Kikuchi, Y. Ichida and H. Honjo, 2005. Vase life of imported anthurium flowers evaluated in Japan in relation to the effects of post importation benzyladenine treatment. HortSci., 40(5):1439- 1443. Furno, F., K.S. Morley, B. Wong, B.L. Sharp, P.L. Arnold, S.M. Howdle, R. Bayston, P.D. Brown, P.D. Winship and H. J. Reid, 2004. Silver nanoparticles and polymeric medical devices, a new approach to prevention of infection. J. Antimicrob. Chemother., 54: 1019-1024. Godarzi, M., 2006. Effects of water and alcoholic extract of thyme on Escherichia coli entrohemorajenic. Res. J. Med. Lolerestan Univ., 8(29): 63-69. Gosh, S., E. Sobudhi and S. Nayak, 2008. Antibacterial assay of Stevia rebaudiana leaf extracts on 10 pathogens. Intl. J. Integrative Biol., (2) 2: 27-31. Grignon, C. and H. Sentenac, 1991. pH and ionic conditions in the apoplast. Annual Rev. Plant Physiol. Plant Mol. Biol., 42:103–128. Hashemabadi, D., 2014. The role of silver nano-particles and silver thiosulfate on the longevity of cut carnation (Dianthus caryophyllus) flowers. J Environ Biol., 35(4):661-666. Hashemabadi, D., A.M. Torkashvand, B. Kaviani, M. Bagherzadeh, M. Rezaalipour and M. Zarchini, 2015. Effect of Mentha pulegium extract and 8-hydroxyl quinoline sulfate to extend the quality and vase life of rose (Rosa hybrida) cut flowers. J. Environ. Biol., 36:215-220. Hassani, R.N. and F. Alimirzaii, 2017. Postharvest foliar application of gibberellic acid and calcium chloride improved vase life and water balance of cut rose flower cv. Velvet. Biological Forum – An Intl. J., 9(1):56-61. Hatami, M., A. Hatamzadeh, M. Ghasemnezhad and M. Ghorbanpour, 2013. The comparison of antimicrobial effects of silver nanoparticles (SNP) and silver nitrate (AgNO3) to extend the vase life of ‘Red Ribbon’ cut rose flowers. Trakia J. Sci., 2: 144-151. He, S., D.C. Joyce, D.E. Irving and J.D. Faragher, 2006. Stem end blockage in cut grevillea, "Crimso Yul-lo" inforescences. Postharvest Biol. Tech., 41:78-84. Hettiarachchi, M.P. and J. Balas, 2005. Postharvest quality of cut anthurium flowers (Anthurium andraeanum L.) after long-distance shipment. Acta Hort., 669:329-336. Ibrahim, S. M.M., L.S. Taha and R. A. Eid, 2011. Extending postharvest life and keeping quality of gerbera cut flowers using some chemical preservatives. J. Appl. Sci. Res., 7(7):1233-1239. Islam, M.S., H. Mehraj; M.Z.K. Roni; S. Shahrin and A.F.M. Jamal Uddin, 2013. Varietal study of anthurium (Anthurium andraeanum) as a cut flower in Bangladesh. J. Bangladesh Academy of Sci., 37(1):103-107. Jowkar, M.M., 2006. Water relations and microbial proliferation in vase solutions of Narcissus tazetta L. cv. ‘Shahla-e-Shiraz’ as affected by biocide compounds. J. Hort. Sci. Biotec., 81:656-660. Jowkar, M.M., M. Kafi, A. Khalighi and N. Hasanzadeh, 2012. Postharvest physiological and microbial impact of hydroxy quinoline citrate as ‘Cherry Brandy’ rose vase solution biocide. Annals of Biol. Res., 3(5):2238-2247. Jowker, M.M., 2005. Effect of different compounds on microbial population of cut ‘Shiraz Narcissus’ vase solution. V. International Postharvest symposium, ISHS, Acta Horti., 682: 1705-1708. Kazemi, M. and A. Ameri, 2012. Response of vase-life carnation cut flower to salicylic acid, silver nanoparticles, glutamine and essential oil. Asian J. Animal Sci., 6: 122-131.

311 Middle East J. Appl. Sci., 7(2): 299-313, 2017 ISSN 2077-4613

Khandaker, M.M., M. Z. M. Rasdi, N.N. Naeimah and N. Mat, 2017. Effects of naphthalene acetic acid (NAA) on the plant growth and sugars effects on the cut flowers Mokara Chark Kuan orchid. Biosci. J. Uberlândia, 33(1): 19-30. Li, H., H. Li , J. Liu, Z. Luo, D. Joyce and S. He, 2017. Nano-silver treatments reduced bacterial colonization and biofilm formation at the stem-ends of cut gladiolus ‘Eerde’ spikes. Postharvest Biol. and Tech., 123:102–111. Liu, J. P., S.G. He, Z.Q. Zhang, J. P. Cao, P.T. Lv, S.D. He, G.P. Cheng and D.C. Joyce , 2009. Nanosilver pulse treatments inhibit stem-end bacteria on cut gerbera cv. Ruikou flowers. Postharvest Biol. Tech., 54(1):59-62. Lü, P., S. He, H. Li, J. Cao and H. Xu, 2010. Effects of nano-silver treatment on vase life of cut rose cv. Movie Star flowers. J. Food Agric Environ., 8 (2): 1118 - 1122. Moradi, P., H. Afshari and A. G. Ebadi, 2012. The effect of benzyl adenine, nano silver, 8- hydroxylquinolin sulfate, and sucrose on longevity improvement and some other quality characteristics of Dianthus cv. Cream Viana cut flower. Ind. J. Sci. Tech., 5:2459-2463. Morones, J. R., J.L. Elechiguerra, A. Camacho, K. Holt, J.B. Kouri, T.J. Ramirez and M.J. Yacaman, 2005. The bactericidal effect of silver nanoparticles. Nanotechnology, 16: 2346-2353. Mortazavi, S.N., F. Bagheri and M. Bahadoran, 2016. Some characteristics of tuberose as affected by pre-harvest application of calcium chloride and gibberellic acid. Adv. Hort. Sci., 30(2): 69-74. Mortazavi, S.N., M. Mohebbi and Y. Sharafi, 2011. Effects of nanosilver and sucrose on vase life of cut rose flower (Rosa hybrida cv. Royal). J. Med. Plants Res., 5:6455-6459. Nemati, S. H., A. Tehranifar; B. Esfandiari and A. Rezaei, 2013. Improvement of vase life and postharvest factors of Lilium orientalis ‘Bouquet’ by silver nano particles. Not Sci Biol., 5(4):490-493. Nemati, S.H., B. Esfandiyari, A. Tehranifar, A. Rezaei and S.J. Ashrafi, 2014. Effect of nano-silver particles on postharvest life of Lilium orientalis cv. ‘Shocking’, Int. J. Postharvest Tech. and Innovation, 4(1):46–53. Nouri, N., V. Abdossi and M. Boojar, 2012. The effect of gibberellic acid and benzyladenine on post harvest quality and vase life of Alstroemeria cultivar Mayfair cut flowers with enzymatic assay. Intel. J. Agric.: Research and Review., 2:1025-1031. Oraee, T., A.A. Zadeh, M. Kiani and A. Oraee, 2011. The role of preservative compounds on number of bacteria on the end of stems and vase solutions of cut Gerbera. J. Orn. & Hort. Plants, 1(3):161- 165. Paull, R. E., 1982. Anthurium (Anthurium andraeanum, Andre) vase life evaluation criteria. HortScience., 17:606-607. Paull, R.E. and T.T.C. Goo, 1985. Ethylene and water stress in the senescence of cut anthurium flowers. J. Amer. Soc. Hort. Sci., 110:84–88. Paull, R.E., N.J. Chen and J. Deputy, 1985. Physiological changes associated with senescence of cut anthurium flowers. J. Amer. Soc. Hort. Sci., 110:156–162. Prashant, P., R.C. Sekhar and K.C.S. Reddy, 2010. Influence of floral preservatives on scape bending, biochemical changes and postharvest vase Life of cut gerbera (Gerbera Jamesonii Bolusex. Hook), Asian J. Horti., 5:1–6. Pun, U.K. and K. Ichimura, 2003. Role of sugars in senescence and biosynthesis of ethylene in cut flowers. JARQ., 37: 219-224. Rafi, Z. N. and A. Ramezanian, 2013. Vase life of cut rose cultivars ‘Avalanche’ and ‘Fiesta’ as affected by nano-silver and s-carvone treatments. South African J. Botany, 86:68–72. Rai. M., A. Yadav and A. Gade, 2009. Silver nanoparticles as a new generation of antimicrobials. Biotech Adv., 27:76-83. Rogers, M. N., 1973. An historical and critical review of postharvest physiology research on cut flowers, Hort. Sci., 8:189-194. Roshani, T., N. Ahmadi and G. Karimzadeh, 2016. Effects of silver nano particles and 1-MCP on postharvest characteristic and activities of enzymes involved in cut carnation flower senescence. Adv. Appl. Sci., p:1-10. Sairam, R. K., B. Vasanthan and A. Arora, 2011. Calcium regulates gladiolus flower senescence by influencing antioxidative enzymes activity. Acta Plant Physiol., 33: 1897-1904.

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SAS, Institute, 2009. SAS/STAT, User`s Guides: Statistics. Vers. 9, SAS. Institute Inc. Cary, N.C., USA. Shah, V. and I. Belozerova, 2009. Influence of metal nanoparticles on the soil microbial community and germination of lettuce seeds. Water, Air, and Soil Pollution 197, 143–148. Singleton, V.L., R. Orthofer and R.M. L. Raventos, 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol. 299:152-178. Solgi, M., M. Kafi, T.S. Taghavi and R. Naderi, 2009. Essential oils and nanoparticles (SNP) as novel agents to extend vase-life of gerbera (Gerbera jamesonii cv., Dune,) flowers. Postharvest Biol. Technol., 53: 155-158. Soumen, M. and N. Roychowdhury, 2005. Effect of pulsing and holding solutions on postharvest life of cut flowers in Anthurium andraeanum Lind. cv. Nitta J. Orn. Hort., 8(3):186-190. Steel, R. G. D. and J. H. Torrie, 1980. Principles and Procedures of Statistics. McGrow Hill Book Co., Inc., New York, p: 377-400. Sven, V. and J.V.G. Jose, 2004. Sucrose loading decreases ethylene responsiveness in carnation (Dianthus caryophyllus cv. White Sim) petals. Postharvest Biol. Tech., 31:305–312. Świder, J. R., E. Skutnik, and A. Jędrzejuk, 2017. The effect of preservatives on water balance in cut clematis flowers. The Journal of Horticultural Science and Biotechnology, 92 (3):1-9. Thwala, M., P.K. Wahome; T.O. Oseni and M.T. Masarirambi, 2013. Effects of floral preservatives on the vase life of orchid (Epidendrum radicans L.) cut flowers. J. Hort. Sci. & Orn. Plants, 5(1):22-29. Van Doorn, W.G., 1997. Water relations of cut flowers. Hort. Rev., 18:1–85. Van Doorn, W.G., 2004. Is petal senescence due to sugar starvation?. Plant Physiol., 134: 35–42. Van Doorn, W.G.; R.R.J. Perik; P. Abadie and H. Harkem , 2011. A treatment to improve the vase life of cut tulips: Effects on tepal senescence, tepal abscission, leaf yellowing and stem elongation. Postharvest Biol. and Tech., 61:56-63. Van Ieperen, W. and A. Van Gelder, 2006. Ion-mediated flow changes suppressed by minimal calcium presence in xylem sap in chrysanthemum and Prunus laurocerasus. J. Exp. Bot., 57: 2743–2750. Vinodh, S., M. Kannan and M. Jawaharlal, 2013. Effect of nanosilver and sucrose on post harvest quality of cut Asiatic lilium cv. Tresor. The Rioscan, 8(3):901-904.

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