J Integr Plant Biol. ›› 2002, Vol. 44 ›› Issue (5): 573-578.

• Research Articles • Previous Articles     Next Articles

Identification of the Rice Vacuolar ATPase B Subunit Gene and Its Expression Pattern Analysis Under Phosphorus Deficiency

XIA Ming, WANG Xiao Bing, LI Hai Bo, WU Ping*   

  • Published:2002-05-15

Abstract:

A vacuolar ATPase (V ATPase) B subunit gene has been cloned and characterized from a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99. Southern analysis of the genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6-12 h) after phosphorus starvation in roots, and lately stage (24-48 h) in leaves. Under phosphorus deficiency, the up regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate phosphorus transport.

水稻液泡ATPase B 亚基基因的克隆及其在低磷
胁迫下的表达特征分析
夏 铭 王小兵 李海波 吴平*

(浙江大学植物科学研究所,杭州310029)

摘要: 利用抑制性扣除杂交(SSH)技术构建水稻(Oryza sativa L.)根系磷饥饿诱导cDNA文库,获得编码液泡ATPase (V-ATPase) B亚基的克隆,通过反转录PCR方法获得该基因的完整序列.该基因编码487个氨基酸,含有一个保守的ATP结合位点,其蛋白分子量为54.06 kD,等电点为4.99.Southern印迹表明,V-ATPase B亚基基因在水稻基因组中以单拷贝形式存在.氨基酸同源性分析发现,V-ATPase B亚基是一个较为保守的蛋白亚基,其序列变化伴随生物的进化过程同步进行.Northern印迹表明,V-ATPase B亚基在水稻根系中受到磷饥饿诱导表达,磷饥饿6~12 h出现表达高峰,而在叶片中表达高峰有所滞后(24~48 h).在缺磷环境条件下,ATPase B亚基可能通过提高其表达量,进而提高质子转运活性,形成跨膜的电化学梯度,为体内储备磷跨液泡膜运输提供能量,从而提高植物体内磷的利用效率及其耐低磷的能力。

关键词: 水稻;液泡ATPase B 亚基;磷饥饿;抑制性扣除杂交(SSH);基因克隆

通讯作者。E-mail: docpwu @ cls.zju.edu.cn,Tel:0571-86971130

Key words: Oryza sativa, vacuolar ATPase B subunit gene, phosphorus starvation, suppression subtractive hybridization (SSH), gene cloning

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