Gao Qi-Fei, Gu Li-Li, Wang Hui-Qin, Fei Cui-Fang, Fang Xiang, Hussain Jamshaid, Sun Shu-Jing, Dong Jing-Yun, Liu Hongtao, Wang Yong-Fei
National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; Institute of Plant Physiology and Ecology, University of Chinese Academy of Sciences, Shanghai 200032, China;
National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;
Proc Natl Acad Sci U S A. 2016 Mar 15;113(11):3096-101. doi: 10.1073/pnas.1524629113. Epub 2016 Feb 29.
In flowering plants, pollen tubes are guided into ovules by multiple attractants from female gametophytes to release paired sperm cells for double fertilization. It has been well-established that Ca(2+) gradients in the pollen tube tips are essential for pollen tube guidance and that plasma membrane Ca(2+) channels in pollen tube tips are core components that regulate Ca(2+) gradients by mediating and regulating external Ca(2+) influx. Therefore, Ca(2+) channels are the core components for pollen tube guidance. However, there is still no genetic evidence for the identification of the putative Ca(2+) channels essential for pollen tube guidance. Here, we report that the point mutations R491Q or R578K in cyclic nucleotide-gated channel 18 (CNGC18) resulted in abnormal Ca(2+) gradients and strong pollen tube guidance defects by impairing the activation of CNGC18 in Arabidopsis. The pollen tube guidance defects of cngc18-17 (R491Q) and of the transfer DNA (T-DNA) insertion mutant cngc18-1 (+/-) were completely rescued by CNGC18. Furthermore, domain-swapping experiments showed that CNGC18's transmembrane domains are indispensable for pollen tube guidance. Additionally, we found that, among eight Ca(2+) channels (including six CNGCs and two glutamate receptor-like channels), CNGC18 was the only one essential for pollen tube guidance. Thus, CNGC18 is the long-sought essential Ca(2+) channel for pollen tube guidance in Arabidopsis.
在开花植物中,花粉管由雌配子体产生的多种吸引剂引导进入胚珠,以释放成对的精细胞进行双受精。花粉管顶端的Ca(2+)梯度对于花粉管导向至关重要,这一点已经得到充分证实,并且花粉管顶端的质膜Ca(2+)通道是通过介导和调节外部Ca(2+)内流来调节Ca(2+)梯度的核心组件。因此,Ca(2+)通道是花粉管导向的核心组件。然而,目前仍缺乏遗传证据来鉴定对花粉管导向至关重要的假定Ca(2+)通道。在此,我们报告,环核苷酸门控通道18(CNGC18)中的点突变R491Q或R578K通过损害拟南芥中CNGC18的激活,导致Ca(2+)梯度异常和强烈的花粉管导向缺陷。CNGC18的花粉管导向缺陷在cngc18-17(R491Q)和转移DNA(T-DNA)插入突变体cngc18-1(+/-)中可被CNGC18完全挽救。此外,结构域交换实验表明,CNGC18的跨膜结构域对于花粉管导向是不可或缺的。另外,我们发现,在八个Ca(2+)通道(包括六个CNGCs和两个类谷氨酸受体通道)中,CNGC18是唯一对花粉管导向至关重要的通道。因此,CNGC18是拟南芥中长期以来寻找的对花粉管导向至关重要的Ca(2+)通道。