麻竹同源异型盒基因DlKNOX1的克隆及功能初步分析

林业科学 ›› 2014, Vol. 50 ›› Issue (2): 56-62.

麻竹同源异型盒基因DlKNOX1的克隆及功能初步分析

刘青¹, 汪玉凤², 赵韩生¹, 陈颖¹, 高志民¹

1. 国际竹藤中心国家林业局竹藤科学与技术重点开放实验室北京 100102;
2. 南京林业大学竹类研究所南京 210037

收稿日期:2013-07-29 修回日期:2013-10-26 出版日期:2014-02-25 发布日期:2014-03-11
基金资助:
“十二五”国家科技支撑计划项目“竹子优良种质选育技术研究与示范”专题“分子辅助育种技术集成与示范”（2012BAD23B0504）。

Molecular Characteristics and Primary Functional Analysis of DlKNOX1 Gene from Dendrocalamus latiflorus

Liu Qing¹, Wang Yufeng², Zhao Hansheng¹, Chen Ying¹, Gao Zhimin¹

1. International Center for Bamboo and Rattan Key Laboratory of Bamboo and Rattan Science and Technology of State Forestry Administration Beijing 100102;
2. Bamboo Research Institute, Nanjing Forestry University Nanjing 210037

Received:2013-07-29 Revised:2013-10-26 Online:2014-02-25 Published:2014-03-11
Contact: 高志民

摘要/Abstract

摘要：

同源异型盒基因在决定植物的形态建成中发挥着重要作用。采用RT-PCR和RACE技术，从麻竹中分离到1个同源异型盒基因，其cDNA全长1 452 bp，编码1个330个氨基酸的蛋白，包含1个KNOX1区、1个KNOX2区、1个ELK区和1个Homeobox区，属于KNOX蛋白，该基因命名为DlKNOX1 （GenBank No. KF709955）。组织表达特异性分析表明，DlKNOX1 在节中的表达量最高，茎和鞘中次之，而在叶片中最低。将DlKNOX1 置于35S启动子控制下，构建到载体pPZP的多克隆位点并转化拟南芥。RT-PCR分析表明，DlKNOX1 已在转基因植株中表达。转基因植株表型变化明显，花期比野生型延迟，果荚着生部位明显发生变化，这意味着DlKNOX1 基因可能参与麻竹形态建成的调控。

关键词: 麻竹, 同源异型盒基因, 组织特异性表达, 异位表达, 形态建成

Abstract:

Homeobox genes play important roles in determining plant morphogenesis. A homeobox gene was isolated from Dendrocalamus latiflorus with RT-PCR and RACE methods. The full length of the cDNA was 1 452 bp which would encode a 330-aa protein containing one KNOX1 domain, one KNOX2 domain, one ELK domain and one Homeobox domain, indicating that it belongs to KNOX protein, and the gene was designed as DlKNOX1 . Tissue specific expression showed that DlKNOX1 expressed in knob with the highest level, followed by in stem and sheath, and the lowest in leaf. DlKNOX1 gene was constructed into the multiple cloning sites of pPZP vector driven by a 35S promoter, and transferred into Arabidopsis thaliana. RT-PCR analysis showed that DlKNOX1 was expressed in the transgenic plants. The phenotype of transgenics showed that flowering time was later than that of wild type, and the pods position changed obviously, which suggested that DlKNOX1 might participate in the regulation of D. latiflorus morphogenesis.

Key words: Dendrocalamus latiflorus, homeobox gene, tissue specific expression, ectopic expression, morphogenesis

中图分类号:

S718.46

刘青, 汪玉凤, 赵韩生, 陈颖, 高志民. 麻竹同源异型盒基因DlKNOX1的克隆及功能初步分析[J]. 林业科学, 2014, 50(2): 56-62.

Liu Qing, Wang Yufeng, Zhao Hansheng, Chen Ying, Gao Zhimin. Molecular Characteristics and Primary Functional Analysis of DlKNOX1 Gene from Dendrocalamus latiflorus[J]. Scientia Silvae Sinicae, 2014, 50(2): 56-62.

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