胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

王丽丽; 赵韩生; 孙化雨; 董丽莉; 娄永峰; 高志民

胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

1.
国际竹藤中心, 竹藤科学与技术重点开放实验室, 北京 100102
基金项目:
国家科技支撑计划专题"分子辅助育种技术集成与示范"(2012BAD23B0504)、国际竹藤中心基本科研业务费专项资金项目"毛竹快速生长期特异转录调控网络的构建及其应用"(1632015008)、国家林业局948项目"小分子RNA分离技术引进"(2011-4-55)。
中图分类号: S795.7

Expression Analysis of miR164b and Its Target Gene PeNAC1 in Phyllostachys edulis under Stress

1.
International Center for Bamboo and Rattan, Key Laboratory on the Science and Technology of Bamboo and Rattan, Beijing 100102, China
CLC number: S795.7

摘要: MicroRNAs(miRNAs)在植物生长发育以及抗逆等过程中起着重要的调控作用。miR164 作为植物特有的miRNA,其主要的靶基因是植物NAC转录因子。为揭示毛竹 miR164 对其靶基因的调控机制,通过茎环引物法和RT-PCR技术,从毛竹(Phyllostachys edulis)中克隆出miR164b成熟序列及其靶基因PeNAC1。序列分析表明miR164b的靶点位于PeNAC1 编码区,RLM-5'RACE PCR产物测序结果证实切割位点位于靶点的第10-11位碱基之间。组织特异性表达分析表明,毛竹 miR164b和PeNAC1在根、茎、叶及鞘中均表达,其中miR164b在根中表达丰度最高,在茎中表达丰度最低;而PeNAC1的表达丰度恰好与miR164b 相反。实时定量PCR结果显示,NaCl(250 mmol·L-1)、低温(4℃)和强光(1 500 μmol·m-2·s-1)处理后毛竹叶片中 miR164b 的表达均明显下调,GA3(100 μmol·L-1)处理后 miR164b表达量明显上调;而在同样处理条件下,PeNAC1的表达恰好呈现出与其完全相反的趋势。由此表明,miR164b对靶基因PeNAC1 具有表达调控作用,可能与毛竹响应非生物胁迫的抗逆过程密切相关,这为利用miRNA开展竹子抗逆分子育种提供了参考。
- 毛竹
- / miRNA
- / NAC转录因子
- / 非生物胁迫
Abstract: MicroRNAs (miRNAs) play important roles in a variety of biological growth processes and stress-resistance responses. As the plant-specific miRNA family, the miR164 mainly targets for NAC transcriptional factors. To reveal the regulatory mechanism for miR164 and its target gene, the mature sequence of miR164b and its targeted sequence of PeNAC1 were isolated respectively from Phyllostachys edulis seedlings by RT-PCR with stem-loop RT primers. The analysis indicated that there was one miR164b complementary site located in the open reading frame region of PeNAC1 mRNA. The sequencing result of RLM-5'RACE confirmed that PeNAC1 was regulated by miR164b through specific cleavage at the site between the 10th and 11th bases. Tissue specific expression of miR164b and PeNAC1 demonstrated that they both expressed in root, stem, leaf and sheath, of which miR164b expressed in root with the highest level and the lowest level in stem, while that of PeNAC1 coincided with miR164b conversely. Real-time quantitative PCR analysis showed that miR164b was down-regulated under the treatments of NaCl (250 mmol·L-1), low temperature (4℃) and high light (1 500 μmol·m-2·s-1), but it was up-regulated obviously under the treatment of GA3 (100 μmol·L-1). Meanwhile, the expression of PeNAC1 showed exactly the opposite trends. It is suggested that miR164b played a regulator role in the expression of PeNAC1, which might be closely related to the resilience process of respond to abiotic stress. This result provides a reference of bamboo molecular breeding for resilience by using miRNA.
- Phyllostachys edulis
- / miRNA
- / NAC transcriptional factor
- / abiotic stress

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胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

Expression Analysis of miR164b and Its Target Gene PeNAC1 in Phyllostachys edulis under Stress

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胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

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Expression Analysis of miR164b and Its Target Gene PeNAC1 in Phyllostachys edulis under Stress

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胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

Expression Analysis of miR164b and Its Target Gene PeNAC1 in Phyllostachys edulis under Stress

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胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

English Abstract

Expression Analysis of miR164b and Its Target Gene PeNAC1 in Phyllostachys edulis under Stress

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