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荔枝(Litchi Chinensis Sonn.)多酚氧化酶基因的克隆、表达分析及其与采后果皮褐变的关系

Cloning and expression analysis of litchi (Litchi Chinensis Sonn.) polyphenol oxidase gene and relationship with postharvest pericarp browning.

作者信息

Wang Jiabao, Liu Baohua, Xiao Qian, Li Huanling, Sun Jinhua

机构信息

Environment and Plant Protection Research Institute, Chinese Academy of Tropical Agriculture Sciences, Haikou, Hainan, China.

出版信息

PLoS One. 2014 Apr 24;9(4):e93982. doi: 10.1371/journal.pone.0093982. eCollection 2014.

Abstract

Polyphenol oxidase (PPO) plays a key role in the postharvest pericarp browning of litchi fruit, but its underlying mechanism remains unclear. In this study, we cloned the litchi PPO gene (LcPPO, JF926153), and described its expression patterns. The LcPPO cDNA sequence was 2120 bps in length with an open reading frame (ORF) of 1800 bps. The ORF encoded a polypeptide with 599 amino acid residues, sharing high similarities with other plant PPO. The DNA sequence of the ORF contained a 215-bp intron. After carrying out quantitative RT-PCR, we proved that the LcPPO expression was tissue-specific, exhibiting the highest level in the flower and leaf. In the pericarp of newly-harvested litchi fruits, the LcPPO expression level was relatively high compared with developing fruits. Regardless of the litchi cultivar and treatment conditions, the LcPPO expression level and the PPO activity in pericarp of postharvest fruits exhibited the similar variations. When the fruits were stored at room temperature without packaging, all the pericarp browning index, PPO activity and the LcPPO expression level of litchi pericarps were reaching the highest in Nandaowuhe (the most rapid browning cultivar), but the lowest in Ziniangxi (the slowest browning cultivar) within 2 d postharvest. Preserving the fruits of Feizixiao in 0.2-μm plastic bag at room temperature would decrease the rate of pericarp water loss, delay the pericarp browning, and also cause the reduction of the pericarp PPO activity and LcPPO expression level within 3 d postharvest. In addition, postharvest storage of Feizixiao fruit stored at 4°C delayed the pericarp browning while decreasing the pericarp PPO activity and LcPPO expression level within 2 d after harvest. Thus, we concluded that the up-regulation of LcPPO expression in pericarp at early stage of postharvest storage likely enhanced the PPO activity and further accelerated the postharvest pericarp browning of litchi fruit.

摘要

多酚氧化酶(PPO)在荔枝果实采后果皮褐变过程中起关键作用,但其潜在机制仍不清楚。在本研究中,我们克隆了荔枝PPO基因(LcPPO,JF926153),并描述了其表达模式。LcPPO cDNA序列长度为2120 bp,开放阅读框(ORF)为1800 bp。该ORF编码一个含有599个氨基酸残基的多肽,与其他植物PPO具有高度相似性。ORF的DNA序列包含一个215 bp的内含子。进行定量RT-PCR后,我们证明LcPPO表达具有组织特异性,在花和叶中表达水平最高。在新采收的荔枝果实果皮中,LcPPO表达水平与发育中的果实相比相对较高。无论荔枝品种和处理条件如何,采后果实果皮中的LcPPO表达水平和PPO活性都表现出相似的变化。当果实无包装室温贮藏时,采后2 d内,南岛无核(褐变最快的品种)荔枝果皮的褐变指数、PPO活性和LcPPO表达水平均达到最高,而紫娘喜(褐变最慢的品种)则最低。室温下将妃子笑果实保存在0.2μm塑料袋中,可降低果皮失水速率,延缓果皮褐变,并在采后3 d内使果皮PPO活性和LcPPO表达水平降低。此外,妃子笑果实采后在4℃贮藏可延缓果皮褐变,同时在采后2 d内降低果皮PPO活性和LcPPO表达水平。因此,我们得出结论,采后贮藏前期果皮中LcPPO表达上调可能增强了PPO活性,进而加速了荔枝果实采后果皮褐变。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c191/3998928/5b15d1961263/pone.0093982.g001.jpg

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