Boavida Leonor C, McCormick Sheila
Plant Gene Expression Center and Plant and Microbial Biology, USDA/ARS and UC-Berkeley, 800 Buchanan Street, Albany, CA 94710, USA.
Plant J. 2007 Nov;52(3):570-82. doi: 10.1111/j.1365-313X.2007.03248.x. Epub 2007 Aug 30.
Despite much effort, a robust protocol for in vitro germination of Arabidopsis thaliana pollen has been elusive. Here we show that controlled temperatures, a largely disregarded factor in previous studies, and a simple optimized medium, solidified or liquid, yielded pollen germination rates above 80% and pollen tube lengths of hundreds of microns, with both Columbia and Landsberg erecta (Ler) ecotypes. We found that pollen germination and tube growth were dependent on pollen density in both liquid and solid medium. Pollen germination rates were not substantially affected by flower or plant age. The quartet1 mutation negatively affected pollen germination, especially in the Ler ecotype. This protocol will facilitate functional analyses of insertional mutants affecting male gametophyte function, and should allow detailed gene expression analyses during pollen tube growth. Arabidopsis thaliana can now be included on the list of plant species that are suitable models for physiological studies of pollen tube elongation and tip growth.
尽管付出了诸多努力，但一直未能找到一种完善的拟南芥花粉体外萌发方案。在此我们表明，以往研究中很大程度上被忽视的控制温度，以及一种简单的优化培养基（固体或液体），能使哥伦比亚和直立型生态型（Ler）的花粉萌发率高于80%，花粉管长度达到数百微米。我们发现，在液体和固体培养基中，花粉萌发和花粉管生长均依赖于花粉密度。花粉萌发率受花朵或植株年龄的影响不大。四重奏1突变对花粉萌发有负面影响，尤其是在Ler生态型中。该方案将有助于对影响雄配子体功能的插入突变体进行功能分析，并应能在花粉管生长过程中进行详细的基因表达分析。拟南芥现在可以被列入适合进行花粉管伸长和顶端生长生理研究的植物物种名单之中。
