| 摘要: |
| 【目的】研究6-磷酸山梨醇脱氢酶基因启动子（S6PDHp）的组织表达特点。【方法】利用Gateway技术构建了S6PDHp与GUS基因的融合表达载体，然后通过农杆菌介导法转化番茄“中蔬5号”，对转基因植株进行PCR检测，对鉴定为阳性的植株进行GUS组织化学染色，验证S6PDHp的组织表达特性。【结果】将S6PDHp-GUS融合表达载体转化番茄，经PCR检测显示，成功获得了转S6PDHp基因的阳性番茄植株。对阳性植株各组织器官的GUS 化学染色和活性检测结果显示，除叶片外许多器官都有很强的GUS活性，且在转基因番茄成熟期植株的茎部活性最高，根部活性最低；叶片从新叶到老叶的各个时期，GUS活性持续降低。【结论】S6PDHp表达在植物衰老过程中降低；且在转基因番茄茎部的GUS活性最强，说明该启动子具有组织表达特异性。 |
| 关键词: 苹果 6-磷酸山梨醇脱氢酶 启动子 转基因 GUS染色 |
| DOI： |
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| 基金项目:国家自然科学基金项目“苹果6 磷酸山梨醇脱氢酶基因启动子的功能分析与应用”（31201600）；国家苹果产业技术体系项目“生物技术与抗性育种”（CARS-28）；国家科技支撑计划项目“主要仁果类果树新品种选育”（2013BAD02B01） |
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| Transformation and transcriptional activity of sorbitol-6-phosphate dehydrogenase promoter of apple |
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YANG Guochan,XIE Yinpeng,MA Fengwang,et al
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| Abstract: |
| 【Objective】This study investigated the tissue-specific expression of sorbitol-6-phosphate dehydrogenase gene promoter (S6PDHp).【Method】The S6PDHp sequence was fused with the GUS reporter gene to construct plant reporter vector.Then,the plant reporter vector was introduced into tomato via Agrobacterium-mediated transformation.PCR detection of the transgenic plants was conducted and tissue-specific expression of S6PDHp in positive plants were validated by GUS histochemical staining.【Result】PCR results showed that the transgenic plants were obtained successfully.In transgenic tomato maturity plant,GUS activity assays indicated that the expression of GUS was highly active in stem while that in root was the lowest.The GUS activity was stronger than that of control plants.New leaf blade to the repressive period of old leaves,GUS activity was reduced.【Conclusion】Promoter expression in plants reduced and GUS activity in transgenic tomato stems was the strongest.The obtained S6PDHp fragment expressed specifically in transgenic tomato. |
| Key words: apple sorbitol-6-phosphate dehydrogenase promoter transgenic GUS staining |
