多酚氧化酶基因NtPPOE差异表达的效应分析

为研究烟草中多酚氧化酶基因的功能,本研究从GenBank中挑选一个烟草多酚氧化酶基因Polyphenol oxidase E(基因登录号:NW_015916967.1),命名为NtPPOE。构建NtPPOE基因的CRISPR载体和超表达载体,分别得到NtPPOE基因抑制/过量表达的转化植株。研究表明,基因敲除植株NtPPOE基因相对表达量降低,PPO活性降低,多酚类物质含量升高;超表达植株NtPPOE基因相对表达量升高,PPO活性升高,多酚类物质含量降低。NtPPOE基因表达量与PPO酶活性呈正相关,与多酚类物质含量呈极显著性负相关,NtPPOE基因的表达影响了烟草多酚代谢。本研究通过分析NtPPOE基因差异表达产生的效应,初步验证了NtPPOE基因的功能,并得到了NtPPOE基因抑制/过量表达的突变植株,为PPO基因功能研究和不同PPO活性品种的育种提供材料基础。

Effect of Differential Expression of Polyphenol Oxidase Gene NtPPOE

To study the function of the polyphenol oxidase gene in tobacco,a tobacco polyphenol oxidase gene polyphenol oxidase E(Gene accession number:NW_015916967.1)was selected from GenBank and named NtPPOE.NtPPOE gene CRISPR vector and overexpression vector were constructed,and transformed plants with NtPPOE gene suppression/overexpression were obtained.Studies have shown that the relative expression of NtPPOE gene in knockout plants is reduced,PPO activity is reduced,and the content of polyphenols is increased;the overexpression of NtPPOE gene is increased,PPO activity is increased,and polyphenols content is reduced.The expression of NtPPOE gene was positively correlated with the activity of PPO enzyme and negatively correlated with the content of polyphenols.The expression of NtPPOE gene affected the metabolism of tobacco polyphenols.In this study,by analyzing the effects of differential expression of the NtPPOE gene,the function of the NtPPOE gene was preliminarily verified,and mutant plants inhibited/overexpressed by the NtPPOE gene were obtained,laying a material foundation for the study of PPO gene function and breeding of different PPO active varieties.