Reduced polyphenol oxidase gene expression and enzymatic browning in potato (Solanum tuberosum L.) with artificial microRNAs.
Reduced polyphenol oxidase gene expression and enzymatic browning in potato (Solanum tuberosum L.) with artificial microRNAs.
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DOI：
10.1186/1471-2229-14-62
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发表时间：
2014-03-11
影响因子：
5.3
通讯作者：
Xiang Y
中科院分区：
文献类型：
--
作者：
Chi M;Bhagwat B;Lane WD;Tang G;Su Y;Sun R;Oomah BD;Wiersma PA;Xiang Y
多酚氧化酶（PPO）通常由多基因家族编码，其导致氧化性布朗宁，这是许多食品中的重要问题。低布朗宁马铃薯以前是通过抑制PPO基因的表达来生产的，但是单个PPO基因异构体对氧化布朗宁过程的贡献是未知的。在这里，我们调查了不同的PPO基因的总PPO蛋白活性的贡献，以及PPO蛋白水平，PPO活性和块茎组织布朗宁潜力之间的相关性，通过抑制所有先前表征的马铃薯PPO基因，单独和组合使用人工microRNA（amiRNA）技术。通过对马铃薯基因组数据库的调查，发现了9个PPO类基因模型，命名为StuPPO 1至StuPPO 9。StuPP 0 1、StuPP 0 2、StuPP 0 3和StuPP 0 4分别与所表征的POTP 1/P2、POT 32、POT 33和POT 72等位。发现较少的EST支持StuPPO 5到StuPPO 8的翻译。StuPPO 9相关EST在病原体感染的马铃薯组织中以显著更高的水平表达。通过抑制StuPPO 1 ~ StuPPO 4基因的单独和联合作用，获得了一系列的布朗宁表型。在转基因马铃薯块茎中，一个或几个PPO基因的下调通常不会引起其他PPO基因的上调，但会导致PPO蛋白水平的降低。不同的PPO基因对块茎组织中总PPO蛋白含量的贡献并不相等，其中StuPPO 2占约55%，其次是StuPPO 1，占约25-30%，StuPPO 3和StuPPO 4占不到15%。PPO蛋白水平、PPO活性与布朗宁势之间存在极显著的正相关关系。低PPO活性和低褐变的马铃薯产生的同时下调StuPPO 2到StuPPO 4，但最大的减少发生时，StuPPO 1到StuPPO 4都被抑制。StuPPO 1 ~ StuPPO 4基因对块茎组织的布朗宁反应有促进作用，但其作用大小不一。不同的PPO基因可以独立调节，反映了它们的多样性功能。我们的研究结果表明，amiRNA可以用来抑制高度保守的多基因家族的密切相关的成员。这种方法也提出了一种新的策略，用于育种低褐变作物使用小的DNA插入。
Polyphenol oxidase (PPO), often encoded by a multi-gene family, causes oxidative browning, a significant problem in many food products. Low-browning potatoes were produced previously through suppression of PPO gene expression, but the contribution of individual PPO gene isoform to the oxidative browning process was unknown. Here we investigated the contributions of different PPO genes to total PPO protein activity, and the correlations between PPO protein level, PPO activity and tuber tissue browning potential by suppression of all previously characterized potato PPO genes, both individually and in combination using artificial microRNAs (amiRNAs) technology. Survey of the potato genome database revealed 9 PPO-like gene models, named StuPPO1 to StuPPO9 in this report. StuPPO1, StuPPO2, StuPPO3 and StuPPO4 are allelic to the characterized POTP1/P2, POT32, POT33 and POT72, respectively. Fewer ESTs were found to support the transcriptions of StuPPO5 to StuPPO8. StuPPO9 related ESTs were expressed at significant higher levels in pathogen-infected potato tissues. A series of browning phenotypes were obtained by suppressing StuPPO1 to StuPPO4 genes alone and in combination. Down-regulation of one or several of the PPO genes did not usually cause up-regulation of the other PPO genes in the transgenic potato tubers, but resulted in reduced PPO protein levels. The different PPO genes did not contribute equally to the total PPO protein content in the tuber tissues, with StuPPO2 accounting for ~ 55% as the major contributor, followed by StuPPO1, ~ 25-30% and StuPPO3 and StuPPO4 together with less than 15%. Strongly positive correlations between PPO protein level, PPO activity and browning potential were demonstrated in our analysis. Low PPO activity and low-browning potatoes were produced by simultaneous down-regulation of StuPPO2 to StuPPO4, but the greatest reduction occurred when StuPPO1 to StuPPO4 were all suppressed. StuPPO1 to StuPPO4 genes contributed to browning reactions in tuber tissues but their effect was not equal. Different PPO genes may be regulated independently reflecting their diversified functions. Our results show that amiRNAs can be used to suppress closely related members of highly conserved multi-gene family. This approach also suggests a new strategy for breeding low-browning crops using small DNA inserts.
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影响因子：
5.1
作者：
HUNT, MD;EANNETTA, NT;STEFFENS, JC
通讯作者：
STEFFENS, JC
影响因子：
7.4
作者：
Thipyapong, P;Joel, DM;Steffens, JC
通讯作者：
Steffens, JC
影响因子：
1.6
作者：
Murata, M;Nishimura, M;Itoh, Y
通讯作者：
Itoh, Y
影响因子：
11.4
作者：
Mallory, AC;Reinhart, BJ;Bartel, DP
通讯作者：
Bartel, DP
影响因子：
6.1
作者：
Coetzer, C;Corsini, D;Tumer, N
通讯作者：
Tumer, N