Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (4): 811-822.doi: 10.3864/j.issn.0578-1752.2020.04.013

• HORTICULTURE • Previous Articles     Next Articles

Influence of Three Enzymes on Oxidation of Ascorbic Acid in Postharvest 'Hayward' and 'Huate' Kiwifruit

Qi FENG1,ChaoZheng LI1,GuiTian GAO1(),You WU1,Yan XIAO1,WuQi ZHAO1,YuShan LEI2   

  1. 1 College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an 710119
    2 Shaanxi Rural Science and Technology Development Center, Xi’an 710054
  • Received:2019-07-09 Accepted:2019-11-30 Online:2020-02-16 Published:2020-03-09
  • Contact: GuiTian GAO E-mail:gaoguitian2006@snnu.edu.cn

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Abstract:

【Objective】This study explored the differences in activities of enzymes and their gene expressions related with oxidation of ascorbic acid (AsA) in postharvest Hayward and Huate kiwifruit to provide a theoretical basis for the oxidation mechanism of AsA in postharvest kiwifruit to regulate fruit ripening and senescence, maintain postharvest quality and extend storage life.【Method】 Hayward and Huate kiwifruit were harvested, and total ascorbic acid (T-AsA), dehydroascorbic acid (DHA), AsA/DHA, and AsA content were measured under the storage conditions of 25℃. Enzyme activity of ascorbic acid oxidase (AO), laccase, ascorbate peroxidase (APX) and gene expression related with AsA oxidation were measured. The relationships between AsA content and the enzyme activity of AO, laccase, APX and gene expression were investigated in the two varieties of kiwifruit.【Result】The content of AsA in Hayward was 86.9 mg/100 g FW at the beginning of harvest, and the loss was about 45% at the end of storage. The content of AsA in Huate was higher at the beginning of harvest (610 mg/100 g FW), and it rose to 886 mg/100 g FW, reached its peak value during the storage, and dropped to 778 mg/100 g FW at the end, which was higher than that on the first day after harvest. The DHA content of the two varieties showed a downward trend throughout the whole storage period, but the DHA content of Hayward was lower than that of Huate. The T-AsA content and AsA content change trend was similar. In the later stage of storage, the AsA/DHA ratio of Hayward was much lower than that of Huate. AO activity was significantly negatively correlated with AsA content of two varieties of kiwifruit, and laccase activity was negatively correlated with AsA content. From the 8th day after harvest, the AO activity of Huate was lower than Hayward, and the laccase activity was lower than Hayward during the whole storage period of Huate kiwifruit. The activity of AO and laccase got the lowest value on the 11th day after harvest; laccase activity reached its highest value in Hayward on the 16th day after harvest. APX had little effect on AsA content, and its activity had no significant correlation with AsA content. Among the three genes in the AO gene family, Achn020161 was a key gene for the oxidative decomposition of AsA, while Achn191341 and Achn316521 had no significant correlation with the oxidation of AsA. Among the three genes in the laccase gene family, Achn007661 and Achn191341 had certain effect on the change of AsA content, while Achn163871 had no significant correlation with the oxidation of AsA. Achn123021, Achn082241 and Achn187071 in the APX gene family had certain effect on the change of AsA content, but they were not key genes for oxidative degradation of AsA.【Conclusion】The high ratio of AsA/DHA played an important role in the accumulation of AsA. AO was the key enzyme on the oxidation of AsA, laccase had certain effect, and APX was not a main enzyme in oxidation of AsA. It’s speculated that Achn020161 in the AO gene family was the key gene for oxidizing AsA, and Achn007661 and Achn19134 in the laccase gene family had certain effects on the oxidation of AsA.

Key words: kiwifruit, Huate, Hayward, ascorbic acid, enzyme activity, gene expression

Table 1

Primers for quantitative real-time PCR"

酶名称 Enzyme name 基因名称 Gene name 上游引物 Forward primer 下游引物 Reverse primer
抗坏血酸氧化酶 AO Achn020161 CGGGTCCTGAACTCTACTGC GCAAGGTAGTCCTGTTTGGA
Achn230551 CCTGTGTGGAACGATGTGGA GCGAGGTAAAGGACAGCGTA
Achn316521 CCAAGTACTCACGGCCACAT AGTGTCTGCTGGGATGAACG
漆酶Laccase Achn007661 CAGTTGTTGCTGTGGATGCT TGAGAAGTGTCGGCTGGTTA
Achn191341 GGTGAAATCTGGGGAGACAA GACTACGACGGATGTGGTTC
Achn163871 CCCAACAAGTGGACAAACATT CCACTTTGGTTGGAGTAAGC
抗坏血酸过氧化物酶 APX Achn123021 CAACTGAAGAGTGCCCGAGA GAATCACCAGCAGCATTGGC
Achn082241 TCCCGGTACTTTCAGAGGTG ACCACAACGACCTCAAGTTC
Achn187071 GGTGGAATGAATGGCTCAAT CACACGCCTCCAGGTTTATA

Fig. 1

Changes of AsA (A), DHA (B), and T-AsA (C) contents of two varieties of kiwifruit during storage “*” and “**” indicate significant differences (P<0.05) and extremely significant (P<0.01) between the two varieties. The same as below"

Fig. 2

Changes of AsA/DHA ratio of two varieties of kiwifruit during storage"

Fig. 3

Changes of AO (A), laccase (B) and APX (C) activities of two varieties of kiwifruit during storage"

Table 2

Relation coefficient among AsA content and related indexes of two varieties of kiwifruit"

总抗坏血酸
T-AsA		 抗坏血酸/脱氢抗坏血酸
AsA/DHA		 抗坏血酸氧化酶
AO		 漆酶
Laccase		 抗坏血酸过氧化物酶
APX
海沃德 Hayward 0.999** 0.839 -0.910* -0.875 0.253
华特 Huate 0.993** 0.834 -0.958* -0.803 -0.500

Fig. 4

Changes of expression levels of related enzyme genes of two varieties of kiwifruit during storage"

Table 3

Relation coefficient among AsA content and genes expression of two varieties of kiwifruit"

海沃德
Hayward		 华特
Huate
Achn020161 -0.922* -0.920*
Achn191341 0.769 0.494
Achn316521 0.02 0.494
Achn007661 -0.947* -0.950*
Achn191341 -0.852 -0.423
Achn163871 -0.325 0.599
Achn123021 -0.84 0.536
Achn082241 -0.390 0.246
Achn187071 -0.803 -0.517
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