Abstract
BackgroundPollen tubes extend rapidly when pollen grains are incubated in defined media. Tube extension requires many critical functions of plant cells including molecular signaling, cytoskeleton remodeling, secretion, and cell wall synthesis. Consequently, pollen tube growth has been established as a way to conduct primary screens of chemical libraries to identify compounds that perturb key cellular processes in plants.ResultsHere we report a simple, inexpensive, rapid and semi-quantitative method for measurement of pollen tube growth in microtiter plates. The method relies on Congo Red binding to pollen tubes and correlates dye fluorescence to tube length.ConclusionsThis method can be used in any laboratory without specialized equipment, and has the potential to enable larger screens as chemical libraries grow and to make chemical screening accessible to researchers building specialized libraries designed to probe pathways in plant biology.
Highlights
Pollen tubes extend rapidly when pollen grains are incubated in defined media
Pollen tubes extend rapidly, reaching lengths in 4 hours that are 10 times the initial grain diameter. This rapid extension occurs by tip growth, a process that relies on many basic cellular functions including actin-myosin-mediated vesicle trafficking [8], calcium gradients and flux [9], GTPase activity [10], kinase signaling [11], and cell wall biosynthesis [12]
Alternative approaches for indirectly quantifying tube length, such as turbidity measurements or ELISAs, have been reported, but each of these require additional processing that reduces their attractiveness for chemical genetic screens [13,14]
Summary
Pollen tubes extend rapidly when pollen grains are incubated in defined media. Tube extension requires many critical functions of plant cells including molecular signaling, cytoskeleton remodeling, secretion, and cell wall synthesis. Small molecules that modulate biological activity have been used in chemical genetic screens to reveal molecular mechanisms underpinning many biological processes in a broad array of organisms [1,2,3]. Pollen tubes extend rapidly, reaching lengths in 4 hours that are 10 times the initial grain diameter. This rapid extension occurs by tip growth, a process that relies on many basic cellular functions including actin-myosin-mediated vesicle trafficking [8], calcium gradients and flux [9], GTPase activity [10], kinase signaling [11], and cell wall biosynthesis [12]. Alternative approaches for indirectly quantifying tube length, such as turbidity measurements or ELISAs, have been reported, but each of these require additional processing that reduces their attractiveness for chemical genetic screens [13,14]
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