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Accepted Manuscript Title: Effects of Co-60 gamma-irradiation and refrigerated storage on quality of Shatang mandarin Author: Ke Zhang Yueye Deng Haohao Fu Qunfang Weng PII: S2213-4530(14)00003-2 DOI: http://dx.doi.org/doi:10.1016/j.fshw.2014.01.002 Reference: FSHW 30 To appear in: Received date: 21-10-2013 Revised date: 7-1-2014 Accepted date: 17-1-2014 Please cite this article as: K. Zhang, Y. Deng, H. Fu, Q. Weng, Effects of Co-60 gamma- irradiation and refrigerated storage on quality of Shatang mandarin, Food Science and Human Wellness (2014), http://dx.doi.org/10.1016/j.fshw.2014.01.002 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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Accepted Manuscript

Title: Effects of Co-60 gamma-irradiation and refrigeratedstorage on quality of Shatang mandarin

Author: Ke Zhang Yueye Deng Haohao Fu Qunfang Weng

PII: S2213-4530(14)00003-2DOI: http://dx.doi.org/doi:10.1016/j.fshw.2014.01.002Reference: FSHW 30

To appear in:

Received date: 21-10-2013Revised date: 7-1-2014Accepted date: 17-1-2014

Please cite this article as: K. Zhang, Y. Deng, H. Fu, Q. Weng, Effects of Co-60 gamma-irradiation and refrigerated storage on quality of Shatang mandarin, Food Science andHuman Wellness (2014), http://dx.doi.org/10.1016/j.fshw.2014.01.002

This is a PDF file of an unedited manuscript that has been accepted for publication.As a service to our customers we are providing this early version of the manuscript.The manuscript will undergo copyediting, typesetting, and review of the resulting proofbefore it is published in its final form. Please note that during the production processerrors may be discovered which could affect the content, and all legal disclaimers thatapply to the journal pertain.

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Effects of Co-60 gamma-irradiation and refrigerated storage 1

on quality of Shatang mandarin 2

Ke Zhang1, Yueye Deng2, Haohao Fu2, Qunfang Weng*3

1 Key Laboratory of Pesticide and Chemical Biology, Ministry of 4

Education,Guangzhou 510642, China; Emails: [email protected] (K. Z.); 5

[email protected] (Y. D.);[email protected] (H. F.). 6

2 The authors who equally contribute this research. 7

* Author to whom correspondence should be addressed; E-Mail: [email protected]

(Q. W.); 9

Tel.: +86-20-8528-0308; Fax: +86-20-8528-0292. 10

Received: / Accepted: Published: 11

12

13

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Abstract: The effectiveness of Co-60 gamma irradiation in controlling citrus red 1

mite (Panonychus citri McGrego) had been proved in our earlier work, however,2

whether it could be used as an alternative method to replace the current way of 3

quarantine treatment against citrus red mites depends on the performances of 4

those effective doses on citrus fruits. This study was conducted to explore the5

effects of Co-60 gamma irradiation on nutrient composition of citrus (Shatang 6

mandarin), selected fruits were divided into different groups and each group were 7

irradiated at 0.0 kGy, 0.2 kGy, 0.3 kGy, 0.4 kGy, 0.5 kGy, 0.6 kGy, respectively, 8

then the treated fruits were stored at temperature of 4 � and the nutrient 9

composition were studied in the following days. The results showed that the 10

shelf-life could be extended when fruits were irradiated at a dose range of 0.2-0.411

kGy, while most unirradiated citrus decayed by 15 days. It also turned out that the 12

citrus irradiated at 0.5 and 0.6 kGy were fully decayed within 45 days of 13

refrigerated storage. Total soluble solids (TSS), total sugar, ascorbic acid (AA),14

and titrable acidity had no significant differences against control during the 15 15

days storage period. Nevertheless, the activities of peroxidase (POD) and 16

superoxide dismutase (SOD) decreased after 15 days; the improvement of storage 17

quality and shelf life may be explained by the change of the protective enzyme 18

activity. As a conclusion, the results of citrus fruit under irradiation at a certain19

dose indicating the potential use of Co-60 gamma irradiation as a safe quarantine 20

treatment.21

Keywords: Shatang mandarin; irradiation; cold storage; nutrient composition22

23

1. Introduction24

Shatang mandarin (Citrus reticulate Blanco) is a kind of characteristic fruit in25

Southeast China, mainly grows in Guangdong Province, South China. It is one of the 26

best commercial fruits of Guangdong Province. Due to the fruit tastes as sweet as 27

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sugar, it’s also called Mitangju. It is cultivated on a large area which accounts for 1

about 80% of citrus planting area in Guangdong Province, due to its reasonably higher2

yield, better quality, greater taste and flavor than those of the other citrus fruits.3

Gamma radiation has been used as a post-harvest food preservation process for 4

many years[1]. Citrus is a seasonal product, which was consumed fresh or processed. 5

Gamma irradiation emerged recently as a possible alternative technology for the citrus6

post-harvest processing, in order to fulfill the requirements of international 7

phytosanitary trade laws. The goal of quarantine or phytosanitary treatments is to 8

prevent the invasion and propagate of regulated pests [2]. Phytosanitary treatments, 9

allow for the destruction or removal of pests or for making the pest reproductively 10

sterile, to achieve the goal of control pest. There are various types of disinfestation 11

treatments including cold, hot water immersion, heated air, methyl bromide 12

fumigation and irradiation [3]. In addition, some of these techniques could help to 13

minimize the loss of quality in terms of flavor, color and nutritional value [4]. Studies 14

have showed that ‘Clemenules’ is a mandarin cultivar highly tolerant to X-ray 15

irradiation and the commercial quality of the fruit was not adversely affected by 16

postharvest quarantine applications that effectively controlled the Medfly [5]. Miller 17

found that grapefruit irradiated at 0.3 kGy resulted in minimal injury to the fruit [6]. 18

Furthermore, studies have demonstrated that ‘Rio Red’ grapefruit exposed to 19

irradiation doses of up to 0.5 kGy did not affect soluble solids, titratable acidity, 20

appearance, and organoleptic quality compared to untreated fruit[7]. Ascorbic acid is 21

water-soluble vitamin, which is present in fresh fruit, especially citrus fruit and 22

vegetable. In addition, ascorbic acid shows antioxidative effects and under certain 23

conditions can protect against oxidativlely induced DNA damage [8]. Khalil reported 24

that citrus fruits with doses of 0.25 and 0.5 kGy alone packed in cellophane bags and 25

stored at room temperature for 42 days, acidity and ascorbic acid values were higher 26

for the oranges irradiated at 0.5 kGy, weight loss decreased and total soluble solid 27

(TSS) increased during storage period[9]. Girennavar studied the influence of E-beam 28

irradiation on bioactive compounds of grapefruits, found that the acidity decreased 29

slightly with an increasing E-beam dose, whereas the total soluble solids increased 30

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and irradiation did not affect the vitamin C content at doses up to 1.0 kGy[ 10].1

Irradiation for postharvest disinfestation has been investigated for various fruits 2

and vegetables and shows great promise in that it sterilizes insects at doses that are 3

low enough not to be detrimental to most fruits and vegetables [11, 12]. The United 4

States Department of Agriculture-Animal and Plant Health Inspection Services 5

(USDA-APHIS), together with other international regulatory bodies, such as the 6

International Atomic Energy Agency (IAEA) and the International Plant Protection 7

Convention (IPPC), have issued guidelines for irradiation treatments to meet export 8

and quarantine restrictions [13].9

The benefits of using irradiation are that the cold treatment allows quality to be 10

maintained, leaves no residue on the product, and reduces the use of fumigants and 11

pallet loads can be treated at a time. Low dose gamma irradiation (1.0 kGy or less) 12

has been shown to control insect pests with little quality loss to fresh produce [14];13

however, energy is imparted into metabolically alive tissues of the commodities, so 14

undesirable damage could be occurred [15]. In 2006, the USDA-APHIS approved 15

generic treatments of 0.15 kGy for fruit flies and 0.40 kGy for all insects except pupa 16

and adult Lepidoptera [16]. However, there are few reports on irradiation quarantine 17

treatments on controlling pest mites. Similarly, the content of health-promoting 18

compounds in citrus fruit may be altered by postharvest treatments such as irradiation. 19

For instance, recent studies showed that irradiation of citrus fruit significantly reduced 20

the total ascorbic acid (TAA) content when irradiation doses were high[10, 17]. 21

However, information is still scarce on the effect of Co60-γ irradiation on nutritional 22

quality of many citrus cultivars.23

The objectives of this research were to evaluate the dose response of quality 24

factors of Shatang mandarin to irradiation at 0.2-0.6 kGy. Meanwhile, we found the 25

irradiation treatment extend the shelf life and delay fruit senescence of the citrus fruit.26

2. Materials and methods27

2.1. Fruits and Reagents28

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Shatang mandarin samples were obtained from a certain supermarket, the citrus 1

fruits were harvested in a commercial orchard in Shaoguan city, Guangdong Province. 2

The uniform size, maturity and fresh citrus were selected.3

Bovine albumin was purchased from Shanghai Fanke Biological Technology Co., 4

Ltd, Guaiacol was purchased from Tianjin Kemiou Chemical reagent Co., Ltd. SOD 5

enzyme kit was purchased from Nanjing Jiancheng Bioengineering Institute, and6

other chemicals and solvents were purchased from Guangzhou Chemical reagent 7

Factory. Water was treated in a water purification system, Unique + UV + UF, 8

Research Scientific Instruments Co., Ltd.9

2.2. Exposure to Co60γ Irradiation10

The experimental material was irradiated by Co60γ-rays at the Furui high-energy 11

Technology Co. Ltd., a Canadian company Nordion Co60γ radiation source, Nansha 12

District, Guangzhou Guangdong Province, China. The samples were divided into six 13

groups to be exposed to different radiation doses (0, 0.2, 0.3, 0.4, 0.5 and 0.6 kGy, 14

and the dose rate was 4 Gy/min, using Fricke dosimeter calibration) with 100 units 15

per group. The groups 2-6 were placed into polyethylene plastic bags and irradiated 16

with 0.2, 0.3, 0.4, 0.5 and 0.6 kGy, respectively. Group 1 was the control sample. The 17

control and the treated samples were then packed in plastic bags and stored at 4 � for 18

45 days.19

2.3. Analysis of major individual quality after Co60γ radiation20

2.3.1.Analysis of weight loss and decay rates21

Each treatment contained 10 fruits and their weight loss were tested. The 22

percentage of weight loss was calculated by following formula:23

[(Fresh weight -Weight at storage interval)/Fresh weight] ×100. 24

Similarly, the decay rate was calculated by (decayed fruits number/total tested 25

fruits number) ×100.26

2.3.2. Total sugar27

Total sugar was determined by saccharimeter (LB32T, Guangzhou Mingrui28

Electronic Technology Co., Ltd.) , measuring refractive index of fruit juices, the 29

concentration of sugar was calculated.30

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2.3.3. Titrable acidity1

1.0 mL fresh juice was added into bidistilled water of 10 mL, titrable acidity was 2

determined according to the AOAC methods[18]. 3

2.3.4. Ascorbic acid4

Ascorbic acid was determined by direct iodine titration. Each 25 ml of the herbal 5

fresh juice was transferred into a 250 ml Erlenmeyer flask. Twenty-five milliliter of 26

N sulfuric acid was added, mixed, diluted with 50 ml of water and 3 ml of starch T.S. 7

was added as an indicator. The solution was directly titrated with 0.1 N iodine 8

previously standardized with primary standard arsenic trioxide. A blank titration was 9

performed prior titration of each sample (n=5). Each ml of 0.1 N iodine is equivalent10

to 8.806 mg ascorbic acid[19].11

2.4. Analysis of protective enzyme activities12

The pulp (1.00 g) of citrus tissues with 10mL 5mM phosphate buffer (pH 7.8 or 13

pH 6.8) in a cold mortar, to grind. The seriflux was then centrifuged at 12,000×g at 4 14

℃ for 15 min. The supernatant was used as the crude enzyme extract. Peroxidase 15

(POD) activity was assayed by measuring the increase in absorbance at 470 nm using 16

4-methylcatechol as a substate prepared in a buffer solution with a pH of 6.8. The 17

reaction was carried out in a 10 mm light path quartz cell. One unit (U) of POD was 18

defined as the amount of enzyme that caused the increase of one absorbance unit (AU) 19

at 470 nm in 10 min. Superoxide dismutase (SOD) activity was assayed by SOD 20

enzyme kit. SOD activity was assessed by measuring the dismutation of superoxide 21

radical generated by xanthine oxidase and hypoxanthine, colormetric analysis. 22

2.5. Statistical analysis23

Statistical analysis was conducted for each of the measured traits by analysis of 24

variance(ANOVA) and the means were separated by Duncan Multiple Range test 25

using the SPSS software, version 18.0 (SPSS, Inc.). In addition, a linear discriminant 26

analysis (LDA) was used to assess the influence of either different storage times or 27

irradiation doses on proximate composition profiles as well as in major individual 28

quality (weight loss, decay rate, total sugar, titrable acidity, ascorbic acid and 29

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protective enzyme activities). All statistical tests were performed at a 5% significance 1

level. All the assays were carried out in triplicate. The results were expressed as mean 2

values with standard deviation (SD).3

4

3. Results and Discussion5

3.1 Physical Properties6

There were little influence on the index of Shatang mandarin when treated at the 7

dose of 0.2-0.4 kGy. The shelf life of Shatang mandarin could be prolonged by dose8

of 0.2-0.4 kGy, and only slight changes of weight losses were observed during the 9

whole experimental process. There was no significant difference between irradiated 10

citrus and the control in weight loss after 7 days, also, weight loss rates of groups 11

treated at 0.2-0.4 kGy, which was no more than 0.01%, were significantly lower than 12

the control after 15, 30 and 45 days by irradiation, respectively(Table 1.), while 13

groups treated by 0.5 and 0.6 kGy were almost decayed, indicating that the irradiation 14

treatment at 0.2-0.4 kGy could prolong the shelf life of Shatang mandarin, and the 15

high dose (0.5-0.6 kGy) would cause damage to fruit.16

Decay rates of groups treated by 0.5 kGy and 0.6 kGy reached 21% and 35% after 17

7 days of treatment, respectively (Table 2.), while, the control and the citrus treated by 18

0.2-0.4 kGy, no rotted fruit was appeared. 15 days after irradiation, 0.5 kGy and 0.619

kGy irradiated citrus decay rates were both more than 50%, while it was only 2.00% 20

and 1.67% in the control group and 0.4 kGy treated group, respectively. There were21

no decay fruit in 0.2 kGy and 0.3 kGy irradiated groups. 30 days after 22

irradiation, the decay rates of 0.5 kGy and 0.6 kGy irradiated groups were more than 23

90%, while the 0.2 kGy and 0.3 kGy treated groups still kept the lowest decay 24

rates, less than 5%, and the decay rates of 0.4kGy irradiated group was 5.3%, as 25

compared to the control 6.7%. The decay rates reached 100% after 45 days26

irradiation, with 0.5 kGy and 0.6 kGy, but they were less than 10% in 0.2 kGy and 0.327

kGy irradiated groups, and the 0.4 kGy irradiated group was lower than that of 28

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control. The irradiation dose below 0.4 kGy (including 0.4 kGy), had a preservative 1

effect on Shatang mandarin, while high doses would cause serious damage to 2

the Shatang mandarin.3

The effect of gamma irradiation on total sugar under different doses was carried 4

out in this study. The results showed that the total sugar content decreased with the 5

extension of storage time (Table 3.). There were no significant differences between 6

treated fruits and the control group in the content of total sugar after 30 7

days. Irradiation dose above 0.5 kGy had cause great damage to the citrus fruit, while 8

0.2-0.4 kGy irradiation treatment had little effect on total sugar content.9

It turned out that citrus fruits treated at doses of 0.2-0.4 kGy had no significant 10

differences in weight loss, soluble solids content, total sugar content and titratable 11

acid content compared with the control group. Fernandes[20] studied the effects of 12

irradiation on chestnut fruits and found that the gamma irradiation doses ≤ 3 kGy did 13

not affect the nutritional and chemical quality of chestnut fruits, which indicated that 14

fruits irradiated at suitable doses would have little harm on its physical properties. 15

According to Farkas[21] , irradiation could reduced storage losses, extended shelf life 16

and/or improved microbiological and parasitological safety of foods. In this study, the 17

similar phenomenon was observed, showing that irradiation doses at proper level18

could prolong the shelf life of Shatang mandarin as well as enhance its appearance 19

quality. We also found that the citrus fruit treated at 0.5 kGy and 0.6 kGy completely 20

decayed in 45 days, suggesting that high dose irradiation treatment had a damaging 21

effect on Citrus fruits, however, at which exactly dose would this happen is still 22

unknown, further studies would be needed to carried out to conform the dosage.23

No significant differences in ascorbic acid between treated fruits and the control 24

were observed in the first 7 days yet it was increased a little bit in control group, 25

which, then sharply decreased on the 30th day and increased again on the 45th26

days. Nevertheless, the ascorbic acid of irradiation treatment groups kept decreasing 27

during the whole experimental period, and this kind of change was much faster in the 28

control than it in irradiation treatment. There were no significant differences between 29

treated groups and the control in ascorbic acid after 15 days and 30 days,30

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respectively.When tested on the 30th day, the ascorbic acid of the treatment groups1

was slightly higher than control but it in groups irradiated at 0.2-0.4 kGy then became 2

lower than control after 45 days treatment. The results illustrated that irradiation 3

treatment doses of 0.2-0.4 kGy on citrus fruits could somehow affect its ascorbic acid 4

content; however, the difference between irradiated samples and the control was not 5

obvious after 30 days of storage at 4 �. 6

The results showed that titratable acid of Shatang mandarin decreased as the 7

storage time extended. There was no significant difference between groups treated by 8

0.2 kGy and 0.3 kGy in titratable acid content when tested on the 7th day and 15th day, 9

but it significantly decreased after 30 and 45 days irradiation treatment in these two 10

groups when it was compared with control (Figure 1.), and the content of titratable 11

acid in groups treated at a dose at 0.3 kGy or higher than it were less than a half of 12

control. It indicated that the irradiation effect on titratable acid content of Shatang 13

mandarin would become serious after 30 days.14

Figure 2 showed that although irradiation treatment had a certain impact on the 15

ascorbic acid content, the ascorbic acid content of irradiation treatment group reduced16

much slower than that of the control. Kaewsuksaeng[22] reported that UV-B treatment 17

induced a gradual increase in citric acid and suppressed the increase of sugar contents 18

during storage. In addition, the ascorbic acid content with or without UV-B treatment 19

decreased during storage, but the decrease in the control was faster than that with 20

UV-B treatment. Others also pointed that irradiation not only affected seed formation 21

in the fruit, but also lowed acidity [23]. From figure 1 we can conclude that titratable 22

acid of Shatang mandarin decreased with the extension of storage time, which 23

revealed that irradiation treatments had significant effect on titratable acid content of 24

Shatang mandarin.25

26

27

Table 1. Weight loss Rate(%)of the citrus fruits after different gamma irradiation 28

treatments.29

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Days in storageSample

7 15 30 45

Control 0.17±0.03a 0.64 ±0.09a 0.96±0.10c 1.22±0.02a

0.2 kGy 0.16±0.05a 0.35±0.01d 0.56±0.03d 0.75±0.02c

0.3 kGy 0.16±0.05a 0.36±0.03d 0.59±0.01d 0.79±0.02bc

0.4 kGy 0.16±0.02a 0.40±0.05cd 0.65±0.05d 0.83±0.004b

0.5 kGy 0.18±0.02a 0.44±0.05bc 1.26±0.09b —

0.6 kGy 0.18±0.02a 0.49±0.01b 1.57±0.07a —

1

Table 2. Decay Rate(%)of the citrus fruits after different gamma irradiation 2

treatments.3

Days in storageSample

7 15 30 45

Control 0.00±0.00c 2.00±1.00c 6.67±1.15c 14.67±2.52b

0.2 kGy 0.00±0.00c 0.00±0.00c 3.00±1.00d 8.00±2.00d

0.3 kGy 0.00±0.00c 0.00±0.00c 3.67±1.15cd 8.33±1.15d

0.4 kGy 0.00±0.00c 1.67±0.58c 5.33±0.58cd 11.33±1.53c

0.5 kGy 21.00±3.61b 55.00±5.00b 92.33±2.52b 100.00±0.00a

0.6 kGy 35.00±5.00a 69.67±4.73a 97.67±2.52a 100.00±0.00a

4

Table 3. Total sugar(g/100 ml)of the citrus fruits after different gamma irradiation 5

treatments.6

Days in storageSample

7 15 30 45

Control 15.42±0.42a 15.11±0.38a 14.99±0.38a 14.65±0.33a

0.2 kGy 15.40±0.42a 15.08±0.38a 14.77±0.37a 14.24±0.29ab

0.3 kGy 15.40±0.42a 14.93±0.37a 14.68±0.35a 14.02±0.26ab

0.4 kGy 15.39±0.42a 14.84±0.35a 14.49±0.32a 13.69±0.29bc

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0.5 kGy 15.37±0.42a 14.66±0.34a 14.16±0.33a —

0.6 kGy 15.07±0.38a 14.15±0.25a 13.98±0.28a —

Note: Date(Table 1, 2 and 3) are presented as the mean±standard deviation of 1

triplicate measurements. Days in storage (7, 15, 30, 45.) mean the days after treatment. 2

Values in the same column with different letters are significantly different (p<0.05).3

4

Figure 1.Chang in content of Titrable acidity after 7, 15, 30, 45 days of different 5

gamma irradiation treated samples and control as compared to time 0 day. Values 6

with an asterisk differ significantly (p < 0.05) from the start of the experiment [CK7

(control); 0.2 kGy treatment, 0.3 kGy treatment, 0.4 kGy treatment, 0.5 kGy treatment and 0.68

kGy treatment, for parts A=7 days after treatment, B=15 days after treatment, C=30 days after 9

treatment, and D=45 days after treatment respectively].10

11

Figure 2.Chang in content of Ascorbic acid of the citrus fruit after 7, 15, 30, 45 12

days of different gamma irradiation treated samples and control as compared to time 0 13

A B

C D

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day. Values with an asterisk differ significantly (p < 0.05) from the start of the 1

experiment [CK(control); 0.2 kGy treatment, 0.3 kGy treatment, 0.4 kGy treatment, 0.5 kGy 2

treatment and 0.6 kGy treatment, for parts A=7 days after treatment, B=15 days after treatment, 3

C=30 days after treatment, and D=45 days after treatment respectively].4

5

6

3.2 Enzyme Activities7

Results of the enzymatic analyses in Figure 3 and 4 illustrated that the SOD 8

enzyme activity of citrus fruits with or without irradiation all increased greatly at the 9

first 7 days, and the control showed the highest activity than other irradiation 10

treatment groups (Figure 3). This may attibute to free radicals induced by 11

irradiation. The SOD activity of all groups dropped to a low level after 15 days, it 12

might because the free radical were cleared by SOD enzyme increased before.13

After 30 days, SOD enzyme activity of all groups bounded back to a relatively higher 14

leve again, in which, the value of 0.4 kGy and 0.5 kGy treatment groups 15

A B

C D

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increased greatly. 45 days after irradiation treatment, 0.5 kGy and 0.61

kGy irradiated citrus fruit decayed completely, and the SOD activity of other groups 2

decreased, and the SOD activity of irradiation treatment groups were lower than the 3

control, showing that irradiated fruit produced more free radicals , however, the 4

treated groups had a larger fluctuations in SOD activity than the control group during 5

the whole experimental process; overall, low dose (�0.4 kGy) irradiation could play 6

a role in delaying Citrus Reticulate Blanco fruit senescence as well as in prolonging 7

the storage shelf. 8

SOD catalyzes the dismutation of superoxide anions to produce hydrogen peroxide, 9

which is then removed by catalase, and the two enzymes are thought to extend food 10

freshness by protecting the integrity of membranes [24]. SOD activity in the citrus fruit 11

increased with or without irradiation treatments, but the activity of the enzymes 12

significantly lower than that in the control throughout 7 days of storage at 4� (Figure 13

3.). It may result of free radicals induced by irradiation . SOD is a primary scavenger 14

for superoxide free radicals, which plays a role in the dismutation of superoxide 15

radicals, whereas catalase (CAT), ascorbate peroxidase (APX) and glutathione 16

reductase (GR) activities would contribute, at least to some extent, to the elimination 17

of hydrogen peroxide[25].18

Studies have been carried out a certain correlation browning of fresh fruits and 19

vegetables with tissue POD, POD through the oxidation reaction can lead to 20

deterioration of the quality of fruits and vegetables. The results of Fig. 2 showed 21

that POD activity of Shatang mandarin fruit increased at first place, then decreased as 22

the storage time extended. The POD activity of all experimental groups increased23

after 7 days irradiation, showing a dose-dependent manner; The POD enzyme activity24

of 0.2-0.4 kGy treatment groups were significantly lower than control after 45 days 25

irradiation, And the 0.5 kGy and 0.6 kGy irradiation groups showed the highest POD 26

activity of the whole experiment process, indicated that high dose 27

irradiation would cause damage to the citrus fruit.28

With regard to the effects on enzyme activity, a study by Falguera et al. [26] found a 29

slightly decrease in the activity of POD in fresh apple juices from Golden, Starking 30

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and Fuji, in UV irradiation treatments, this value slightly decreased during the 1

experiment. Meanwhile, in the juice from King David the loss was 70.0%. In this 2

study, a decrease of POD activity was also observed (Figure 4), Shatang 3

mandarin, low acidity, contained ascorbic acid isn't enough to inhibit the activity of 4

POD for long time, a large number of high active POD will lead to deterioration 5

of the quality. The reason of gamma irradiation to extend the shelf life of the 6

fruit might be due to the influence of the basic metabolism of the fruit, 7

suppressing respiratory enzyme activity, inhibiting the release of CO2 and ethylene 8

production, resulting in delayed ripening and senescence; secondly, irradiation could 9

kill microbial spoilage of fruit, and irradiation played a role in preservation. And it 10

suggested that the SOD and POD enzymes played important roles in the citrus fruit 11

senescence, while the mechanism needed to be further explored.12

13

Figure 3.Effect of Co60γ irradiation on the superoxide dismutase (SOD) of the citrus 14

treated with or without irradiation after 7, 15, 30, 45 days at different dose during 15

storage at 4 �.16

17

18

Figure 4.Effect of Co60γ irradiation on the peroxidase (POD) activity of the citrus 19

treated with or without irradiation after 7, 15, 30, 45 days at different dose during 20

storage at 4 �.21

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1

2

The results of effects of Co-60 gamma-irradiation on the quality index and the 3

protective enzyme activities of the citrus fruit during storage. Our studies showed that 4

low dose irradiation (0.2-0.4 kGy) treatments could prolong the shelf life of the citrus 5

fruit and these doses would slightly affect the quality of citrus fruits with an 6

acceptable level. These results showed that certain Co-60 gamma-irradiation could be 7

a promising safety method to control insect pests and extend the shelf life and delay 8

senescence of other kinds of fruits and vegetables.9

4. Conclusions10

Our results suggested that irradiation at the dose range of 0.2-0.4 kGy and in 11

combination with refrigerated storage is an effective post-harvest technique in 12

mitigating the risk of pest and decay of quarantined fruit.13

Acknowledgments14

The researchers gratefully acknowledge the grants from the International Atomic 15

Energy Agency under Research Contract No.15630.16

Conflicts of Interest17

The authors declare no competing financial interest.18

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