甘薯紫外光受体IbUVR8基因克隆及表达特性分析
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国家甘薯产业技术体系(CARS10);福建省人民政府中国农业科学院农业高质量发展超越“5511”协同创新工程(KXXYJBG0039);福建省属公益类科研院所基本科研专项(2020R1031008);福建省农科院农业科技创新联盟专项(CXLM2021001)


Cloning and Expression Analysis of UVB Photoreceptor IbUVR8 in Sweet Potato
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    摘要:

    UVR8是植物唯一的UVB紫外光受体,为深入研究甘薯UVR8的功能,该研究以叶菜型甘薯‘福菜薯23号’为试验材料,采用CODEHOP结合RACE技术,设计简并引物,克隆得到甘薯UVR8全长cDNA序列,并命名为IbUVR8,利用qRTPCR分析IbUVR8在非生物胁迫下的表达特性,并通过原核表达获取IbUVR8蛋白,为IbUVR8基因在甘薯花青素合成调控作用奠定理论基础。结果表明:(1)成功克隆获得甘薯IbUVR8基因(NCBI登录号KT749986),该基因全长开放阅读框为1 329 bp,共编码了441个氨基酸,相对分子量为47 kD,理论等电点为5.72。(2)序列对比和进化分析结果显示,IbUVR8与番茄、马铃薯UVR8蛋白序列相似性高,进化相对保守。(3)qRTPCR结果显示,IbUVR8基因在甘薯中具有组织特异性,表达量为叶片>茎>根;在低温、干旱和NaCl等非生物胁迫下,IbUVR8基因在3种不同叶色甘薯中均为上调表达;在激素GA3和重金属Cu2+胁迫下,IbUVR8基因在不同甘薯材料的表达趋势差异显著。(4)原核表达获得大小约为80 kD的IbUVR8GST融合蛋白。研究认为,甘薯IbUVR8基因受低温、干旱和NaCl等胁迫影响,推测IbUVR8可能在甘薯抗逆胁迫信号通路中发挥调控作用。

    Abstract:

    UVR8 is the only UVB photoreceptor in plants. In order to further study the function of UVR8 in sweet potato, this study took leafy sweet potato ‘Fucaishu 23’ as the experimental material, and used CODEHOP combined with RACE technology to clone the fulllength cDNA sequence of UVR8 from sweet potato, and named IbUVR8. The expression characteristics of IbUVR8 under abiotic stress were analyzed by qRTPCR, and the IbUVR8 protein was obtained by prokaryotic expression,which lay a theoretical foundation for the regulation of IbUVR8 on anthocyanin biosynthesis in sweet potato. The results showed: (1) the sweet potato IbUVR8 gene was successfully cloned. The fulllength open reading frame of the gene was 1 329 bp, encoding 441 amino acids, with a relative molecular weight of 47 kD and a theoretical isoelectric point of 5.72. The NCBI accession number was KT749986. (2) The results of sequence comparison and evolution analysis showed that IbUVR8 had high sequence similarity with tomato and potato UVR8 protein and was relatively conservative in evolution. (3) The results of qRTPCR showed that IbUVR8 was tissuespecific in sweet potato, and the expression level was leaf>stem>root. Under abiotic stresses such as low temperature, drought and NaCl, the expression of IbUVR8 was upregulated in three different leafcolored sweet potatoes. Under the stress of hormone GA3 and heavy metal Cu2+, the expression of IbUVR8 was significantly different among different sweet potato varieties. It was speculated that the expression of this gene was affected by low temperature, drought and NaCl stress. (4) IbUVR8GST fusion protein with a size of about 80 kD was obtained by prokaryotic expression. The results indicated that IbUVR8 was affected by low temperature, drought and NaCl stress, which suggested that IbUVR8 may play a regulatory role in stress resistance signal pathway in sweet potato.

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李国良,林赵淼,张 鸿,等.甘薯紫外光受体IbUVR8基因克隆及表达特性分析[J].西北植物学报,2022,42(12):2013-2020

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  • 在线发布日期: 2023-01-15
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