Messerli M A, Créton R, Jaffe L F, Robinson K R
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
Dev Biol. 2000 Jun 1;222(1):84-98. doi: 10.1006/dbio.2000.9709.
Pollen tubes grown in vitro require an intracellular tip-high gradient of Ca2+ in order to elongate. Moreover, after about 2 h in vitro both the tip Ca2+ and the elongation rate of lily tubes begin to oscillate regularly with large amplitudes. This raises the question of the phase relation between these two oscillations. Previous studies lacked the temporal resolution to accurately establish this relationship. We have studied these oscillations with a newly developed, high temporal resolution system and the complementary use of both luminescent and fluorescent calcium reporters. We hereby show that the periodic increases in elongation rate during oscillatory growth of Lilium longiflorum pollen tubes clearly precede those in subtip calcium and do so by 4.1 +/- 0.2 s out of average periods of 38.7 +/- 1.8 s. Also, by collecting images of the light output of aequorin, we find that the magnitude of the [Ca2+] at the tip oscillates between 3 and 10 microM, which is considerably greater than that reported by fluorescent indicators. We propose an explanatory model that features cyclic growth and secretion in which growth oscillations give rise to secretion that is essential for the subsequent growth oscillation. We also critically compile data on L. longiflorum stylar growth rates, which show little variation from in vitro rates of pollen tubes grown in optimal medium.
体外生长的花粉管需要细胞内钙离子尖端高梯度才能伸长。此外,在体外培养约2小时后,百合花粉管的尖端钙离子浓度和伸长率开始有规律地大幅振荡。这就引出了这两种振荡之间相位关系的问题。以往的研究缺乏时间分辨率来准确确定这种关系。我们使用新开发的高时间分辨率系统以及发光和荧光钙报告基因的互补应用来研究这些振荡。我们在此表明,在麝香百合花粉管振荡生长期间伸长率的周期性增加明显先于亚尖端钙离子浓度的增加,平均周期为38.7±1.8秒,前者比后者提前4.1±0.2秒。此外,通过收集水母发光蛋白光输出的图像,我们发现尖端的[Ca2+]浓度在3至10微摩尔之间振荡,这比荧光指示剂报告的浓度要高得多。我们提出了一个解释模型,其特征是循环生长和分泌,其中生长振荡引发分泌,而分泌对于随后的生长振荡至关重要。我们还严格整理了麝香百合花柱生长速率的数据,这些数据显示与在最佳培养基中生长的花粉管的体外生长速率几乎没有差异。