* To whom correspondence should be addressed.
Received November 28, 2002; Revision received May 12, 2003
The role of Cl- transport across the plasma membrane was studied in an early step of pollen grain germination in tobacco Nicotiana tabacum L. The Cl- channel blockers, 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and niflumic acid, completely suppress the germination with IC50 ~ 8 µM. At this concentration NPPB reduces the rate of Cl- efflux out of pollen grain by 1.8-fold in the interval 5-12 min, and niflumic acid reduces the rate 1.2-fold. 4,4´-Diisothiocyanatostilbene-2,2´-disulfonic acid, a known inhibitor of Cl- channels and antiporters, completely suppresses germination as well (IC50 = 240 µM), but has no effect on the rate of Cl- efflux. Inhibitors of chloride co-transporters, such as furosemide, bumetanide, and bis(1,3-dibutylbarbituric acid)pentamethine oxonol, suppress the germination by less than 50%. This set of data suggests that NPPB-sensitive anion channels are involved in the activation of pollen grains in the early stage of germination.
KEY WORDS: anion channel, anion co-transporter, NPPB, DIDS, furosemide, bumetanide, DiBAC4(5), pollen germination