拟南芥CBL9的质膜定位对花粉管顶端生长的影响

doi:10.13560/j.cnki.biotech.bull.1985.2018-1093

生物技术通报 ›› 2019, Vol. 35 ›› Issue (5): 58-63.doi: 10.13560/j.cnki.biotech.bull.1985.2018-1093

拟南芥CBL9的质膜定位对花粉管顶端生长的影响

周利明, 房玮

华北理工大学生命科学学院,唐山 063210

收稿日期:2018-12-26 出版日期:2019-05-26 发布日期:2019-05-23
作者简介:周利明,男,博士,研究方向：植物细胞信号传导;E-mail：zhouliming363@163.com
基金资助:
国家自然科学基金项目（31400168）,河北省自然科学基金项目（C2014209216,C2018209112）,国家重点实验室开放课题（SKLECRA2013OFP15）

Effects of Plasma Membrane Localization of Arabidopsis thaliana CBL9 on the Growth of Pollen Tube Tip

ZHOU Li-ming, FANG Wei

College of Life Sciences,North China University of Science and Technology,Tangshan 063210

Received:2018-12-26 Published:2019-05-26 Online:2019-05-23

摘要/Abstract

摘要： 旨在揭示拟南芥CBL9（Calcineurin B-like protein 9）在花粉管顶端生长中的作用。基于花粉管瞬时表达体系,通过基因枪技术将带有黄色荧光蛋白（YFP）标签的CBL9在烟草花粉中进行瞬时表达,观察其亚细胞定位及过表达表型。针对CBL9的N端酰基化的保守位点,通过点突变技术构建CBL9G2A突变体,对比研究其表型及定位的改变情况。结果显示CBL9-YFP定位于花粉管顶端质膜及颗粒状细胞器上,CBL9过量表达可以引起显著的花粉管去极化生长。而CBL9G2A-YFP则表现出非特异性的胞内弥散定位,同时CBL9G2A过量表达没有显著影响花粉管的顶端生长。CBL9的N端酰基化位点是CBL9质膜定位的重要因素,而正确定位的CBL9才能发挥调控花粉管生长的相关功能。

关键词: 拟南芥, 花粉管, 顶端生长, 钙离子, 类钙调神经磷酸酶B亚基

Abstract: This work is aimed to understand the role of Arabidopsis thaliana CBL9（calcineurin B-like protein 9）in the growth of pollen tube tip. Based on the transient expression system,the CBL9 with YFP-tag were transiently expressed in tobacco pollen grains by gene gun,and the overexpression phenotypes and subcellular localization patterns of CBL9-YFP were observed. At the conservative sites for N-terminal acylation of CBL9,a mutant version of CBL9,CBL9G2A was constructed via overlap-PCR-mediated site mutagenesis,and the changes in their phenotypes and localization were compared and studied. The results demonstrated that CBL9-YFP was localized to plasma membrane（PM）of growing pollen tubes and granular organelles. CBL9 overexpression induced a severe loss of growth polarity,leading to a short tube with the swelling tip. CBL9G2A was evenly distributed in cytosol of pollen tubes,and its overexpression did not cause the depolarization growth of pollen tubes. These results suggested that N-terminal acylation site of CBL9 is an important factor in correct targeting to PM,which is required for its function in polarized growth of pollen tubes.

Key words: Arabidopsis thaliana, pollen tube, tip growth, calcium, calcineurin B-like protein

周利明, 房玮. 拟南芥CBL9的质膜定位对花粉管顶端生长的影响[J]. 生物技术通报, 2019, 35(5): 58-63.

ZHOU Li-ming, FANG Wei. Effects of Plasma Membrane Localization of Arabidopsis thaliana CBL9 on the Growth of Pollen Tube Tip[J]. Biotechnology Bulletin, 2019, 35(5): 58-63.

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拟南芥CBL9的质膜定位对花粉管顶端生长的影响

Effects of Plasma Membrane Localization of Arabidopsis thaliana CBL9 on the Growth of Pollen Tube Tip

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