Palanivelu Ravishankar, Preuss Daphne
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL 60637, USA.
BMC Plant Biol. 2006 Apr 5;6:7. doi: 10.1186/1471-2229-6-7.
Pollen tubes deliver sperm after navigating through flower tissues in response to attractive and repulsive cues. Genetic analyses in maize and Arabidopsis thaliana and cell ablation studies in Torenia fournieri have shown that the female gametophyte (the 7-celled haploid embryo sac within an ovule) and surrounding diploid tissues are essential for guiding pollen tubes to ovules. The variety and inaccessibility of these cells and tissues has made it challenging to characterize the sources of guidance signals and the dynamic responses they elicit in the pollen tubes.
Here we developed an in vitro assay to study pollen tube guidance to excised A. thaliana ovules. Using this assay we discerned the temporal and spatial regulation and species-specificity of late stage guidance signals and characterized the dynamics of pollen tube responses. We established that unfertilized A. thaliana ovules emit diffusible, developmentally regulated, species-specific attractants, and demonstrated that ovules penetrated by pollen tubes rapidly release diffusible pollen tube repellents.
These results demonstrate that in vitro pollen tube guidance to excised A. thaliana ovules efficiently recapitulates much of in vivo pollen tube behaviour during the final stages of pollen tube growth. This assay will aid in confirming the roles of candidate guidance molecules, exploring the phenotypes of A. thaliana pollen tube guidance mutants and characterizing interspecies pollination interactions.
花粉管在响应吸引和排斥信号的过程中穿过花组织后传递精子。玉米和拟南芥的遗传分析以及蓝猪耳的细胞消融研究表明,雌配子体(胚珠内的7细胞单倍体胚囊)和周围的二倍体组织对于引导花粉管到达胚珠至关重要。这些细胞和组织的多样性以及难以接近性使得表征引导信号的来源及其在花粉管中引发的动态反应具有挑战性。
在这里,我们开发了一种体外试验来研究花粉管对切除的拟南芥胚珠的引导作用。使用该试验,我们辨别了后期引导信号的时间和空间调节以及物种特异性,并表征了花粉管反应的动态变化。我们确定未受精的拟南芥胚珠会释放可扩散的、受发育调节的、物种特异性的吸引剂,并证明被花粉管穿透的胚珠会迅速释放可扩散的花粉管排斥剂。
这些结果表明,体外花粉管对切除的拟南芥胚珠的引导有效地概括了花粉管生长最后阶段的许多体内花粉管行为。该试验将有助于确认候选引导分子的作用,探索拟南芥花粉管引导突变体的表型,并表征种间授粉相互作用。