干旱和盐胁迫中大麦实时定量PCR内参基因的筛选
Reference Genes Screening for Quantitative Real-Time PCR in Barley under Drought and Salt Stress
查看参考文献27篇
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文摘
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为筛选大麦中干旱和盐胁迫条件下适宜的实时定量PCR内参基因,我们对栽培大麦23-19和野生大麦4-3进行了3种不同的胁迫处理:不浇水自然干旱、30% PEG干旱胁迫、0.4 mol/L NaCl盐胁迫,用内参分析软件GeNorm和NormFinder分析比较了6种常用内参基因ubiquitin、a-tubulin 2、Actin、ARF1、EF1a、 SAM在不同组织部位的表达稳定性,并以筛选到的稳定内参分析了胁迫诱导表达基因DHN1的表达情况。结果显示:筛选内参基因时,将所有实验数据综合在一起分析,未能找到稳定表达的内参基因及其组合;按根、茎、叶进行内参基因的筛选发现不同的组织部位、在不同的胁迫处理下,稳定表达的内参基因不完全相同;叶片中内参基因的表达稳定性最为一致,ubiquitin、Actin和ARF1在3种不同的胁迫处理下均有较为稳定的表达。以筛选到的内参基因ubiquitin、Actin和ARF1分析DHN1的表达情况,结果与植物的生长表现吻合。研究结果表明同时考虑太多的变量组合(如不同胁迫,不同组织)不易筛选到稳定表达的内参基因,建议在进行实时定量PCR研究之前根据具体的实验条件先对内参基因进行筛选。叶片适宜作为组织材料、ubiquitin、 Actin和ARF1适宜作为内参基因组合对干旱和盐胁迫下野生大麦和栽培大麦进行实时定量PCR研究。 |
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其他语种文摘
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To screen the most reliable reference genes for barley quantity real-time PCR, 6 different candidate references (ubiquitin, a-tubulin 2, Actin, ARF1, EF1a, SAM) were studied. Barley 23-19 and wild barley 4-3 were treated with different stresses (no water supply, 30%PEG, 0.4 mol/L NaCl). Software of GeNorm and NormFinder were used to evaluate the expression stability of different genes. No reliable references genes can be selected if combined and analyzed all the samples together. While analyze different tissues (root, shoot, and leaves) and different stress separately, different reliable references genes can be screened. The expression patterns of DHN1 were used to confirm the reliability of reference genes. This study recommend ubiquitin, Actin and ARF1 as the reliable reference gene in barley quantity real-time PCR research under drought and salt stress and leaves can be used as the suitable tissues. |
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来源
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分子植物育种
,2016,14(11):3093-3101 【核心库】
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DOI
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10.13271/j.mpb.014.003093
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关键词
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大麦
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干旱胁迫
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盐胁迫
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内参基因
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地址
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1.
南方科技大学, 深圳生物系细胞微环境重点实验室, 深圳, 518055
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中国科学院西北生态环境资源研究院, 兰州, 730000
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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1672-416X |
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学科
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农作物 |
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基金
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国家自然科学基金青年科学基金
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文献收藏号
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CSCD:5869464
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