Journal of Nuclear Agricultural Sciences ›› 2019, Vol. 33 ›› Issue (10): 1905-1912.DOI: 10.11869/j.issn.100-8551.2019.10.1905

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Cloning and Analysis of the Flowering-related Gene,BoAG-like in Bambusa oldhamii

WEI Hantian, SHI Yan, LI Ling, LIN Xinchun*   

  1. Zhejiang Provincial Collaborative Innovation Center for Bamboo Resources and High-efficiency Utilization/ State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Hangzhou, Zhejiang 311300
  • Received:2018-04-09 Revised:2018-07-24 Online:2019-10-10 Published:2019-07-31

绿竹花发育相关BoAG-like基因的克隆与表达特性分析

魏涵天, 时燕, 李玲, 林新春*   

  1. 浙江农林大学省部共建亚热带森林培育国家重点实验室/浙江省竹资源与高效利用协同创新中心, 浙江 杭州 311300
  • 通讯作者: * 林新春,男,教授,主要从事植物生物技术研究。E-mail:lxc@zafu.edu.cn
  • 作者简介:魏涵天,男,主要从事生物技术研究。E-mail:1620753918@qq.com
  • 基金资助:
    省部共建亚热带森林培育国家重点实验室自主课题(ZY20180203),浙江省农业(林木)新品种选育重大科技专项(2016C02056-8)

Abstract: The AG gene belongs to the MADS-box family and plays an important role in the determination of floral meristem characteristics and floral organ development. To analysis the gene expression pattern of AG in bamboo, a complete cDNA sequence named BoAG-like was cloned from in vitro flower buds of Bambusa oldhamii by using the rapid amplification of cDNA ends technology. The results showed that: BoAG-like gene was 792 bp in length, prossessed AGI and AGII regions which were unique to the AG gene. The BoAG-like protein shared more than 80% similarity with the AG-like homologous proteins including OsMADS58 of Oryza sativa, ZAG1 of Zea mays, and PhMADS3 of Phyllostachys edulis, all belonged to the euAG line of the C gene. Tertiary structure of BoAG-like protein showed that this protein displayed a similar tertiary structure with OsMADS58 of Oryza sativa and BdMADS18 of Brachypodium distachyon, but still with significant differences. RT-qPCR analysis revealed that the expression level of BoAG-like in flower organs was much higher than that in vegetative organs, and increased with the development of flower organs. The expression level of BoAG-like gene in B. oldhamii was significantly different among different parts of flower buds, with the highest level in the pistil and the lowest in the lemma. This study provides a guideline for further research of the molecular mechanism of bamboo flower development.

Key words: Bambusa oldhamii, AG\like, flower development, gene cloning, expression analysis

摘要: AG基因属于MADS-box家族,在决定花分生组织特性和花器官发育中起着重要的作用。为阐明竹类植物AG基因的表达特性,本研究以绿竹开花试管苗花芽为植物材料,采用cDNA末端快速扩增技术(RACE),获得了完整的cDNA序列,命名为BoAG-like。序列分析结果表明,BoAG-like长度为792 bp,具有AG基因特有的AG Ⅰ区和AG Ⅱ区;BoAG-like与水稻OsMADS58、玉米ZAG1和毛竹PhMADS3的AG-like同源蛋白所编码的氨基酸序列同源性达到80%以上,均属于C类基因的eu-AG系;蛋白的三级结构预测结果表明,BoAG-like与水稻OsMADS58和二穗短柄草BdMADS18最为接近,但也存在明显差异;实时荧光定量PCR结果表明,BoAG-like基因在花器官的表达量明显高于营养器官的表达量,且其表达量随着绿竹花发育进程呈上升趋势;BoAG-like的表达量在绿竹花芽不同部位存在显著差异,其中在雌蕊中最高,外稃中最低。本研究结果为进一步深入研究竹子花发育的分子机制奠定了一定的理论基础。

关键词: 绿竹, AG-like, 花发育, 基因克隆, 表达分析