向日葵E3泛素连接酶基因的分析定位和表达鉴定
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内蒙古农牧业青年创新基金(2017QNJJNO3);


Subcellular Localization Analysis and Expression Verification of E3 Ubiquitin Ligase Gene HERC2 of Sunflower (Helianthus annuus L.)
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    摘要:

    该研究利用前期获得的向日葵耐盐相关基因E3泛素连接酶基因序列(HERC2),构建瞬时表达载体Cam35SHERC2GFP,采用基因枪法转化洋葱表皮细胞进行亚细胞定位;采用RTPCR技术,分析盐胁迫下HERC2在耐盐品种P50和盐敏感品种P29根、下胚轴和叶中的表达差异;构建HERC2植物表达载体pPZP221HERC2,采用农杆菌介导法将HERC2导入烟草,进行耐盐功能验证。结果表明:(1)HERC2蛋白定位在细胞膜、细胞质和细胞核中。(2)受到NaCl胁迫后,HERC2基因在耐盐品种P50和盐敏感品种P29中均上调表达,但耐盐品种中的表达量较高。(3)HERC2基因的表达,能够提高转基因烟草的耐盐性。该研究结果为进一步解析向日葵对盐胁迫的响应机制,以及耐盐新品种的选育奠定了基础。

    Abstract:

    In this study, utilizing the salt tolerant gene E3 ubiquitin ligase HERC2 sequence in sunflower which acquired previously, we constructed the transient expression vector Cam35SHERC2GFP. Subcellular localization was carried out by transforming onion epidermal cells with particle bombardment. The expression analysis, that of root, hypocotyl and leaf between the salttolerant hybrid P50 and the saltsensitive hybrid P29 under 120 mmol·L-1 NaCl stress, was done by RTqPCR. Plant expression vector pPZP221HERC2 was constructed. HERC2 was transferred into tobacco to verify salttolerant functionality by Agrobacteriummediated. The results showed that: (1) subcellular localization showed that HERC2 protein expressed in cell membrane, cytoplasm and nucleus; (2) the expression of HERC2 gene was upregulated between the salttolerant hybrid P50 and saltsensitive hybrid P29 under NaCl stress, but the expression quantity of the former is higher than that of the latter; (3) the salt tolerance of transgenic HERC2 tobacco was stronger than that of the wild type tobacco. The study laid a foundation for analyzing of salt tolerance mechanism in sunflower and breeding salttolerant cultivar.

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张艳芳,孙瑞芬,郭树春,等.向日葵E3泛素连接酶基因的分析定位和表达鉴定[J].西北植物学报,2018,38(2):211-218

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  • 在线发布日期: 2018-03-23
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