Genetic evidence as long as essential calcium transporters in pollen growth in addition to fertilization. Sabine Frietsch 1,3, Shawn M. Romanowsky 1,2, Morton Schiøtt 4, Michael G. Palmgren 4, Jeffrey F. Harper 1,2 sfrietsch@uscd.edu in addition to jfharper@unr.edu

Genetic evidence as long as essential calcium transporters in pollen growth in addition to fertilization. Sabine Frietsch 1,3, Shawn M. Romanowsky 1,2, Morton Schiøtt 4, Michael G. Palmgren 4, Jeffrey F. Harper 1,2 sfrietsch@uscd.edu in addition to jfharper@unr.edu www.phwiki.com

Genetic evidence as long as essential calcium transporters in pollen growth in addition to fertilization. Sabine Frietsch 1,3, Shawn M. Romanowsky 1,2, Morton Schiøtt 4, Michael G. Palmgren 4, Jeffrey F. Harper 1,2 sfrietsch@uscd.edu in addition to jfharper@unr.edu

Carnes, Jennifer, Managing Editor has reference to this Academic Journal, PHwiki organized this Journal Genetic evidence as long as essential calcium transporters in pollen growth in addition to fertilization. Sabine Frietsch 1,3, Shawn M. Romanowsky 1,2, Morton Schiøtt 4, Michael G. Palmgren 4, Jeffrey F. Harper 1,2 1 The Scripps Research Institute, Cell Biology, 10550 N. Torrey Pines Rd, 92037 La Jolla, CA, USA 2 University of Nevada, Biochemistry, 1664 North Virginia Str, 89557 Reno, CA, USA 3 University of Cali as long as nia, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0116 4 The Royal Veterinary in addition to Agricultural University, Plant Biology, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark sfrietsch@uscd.edu in addition to jfharper@unr.edu Introduction: Calcium dynamics are thought to play a central role in pollen development, as evidenced by pharmaco-chemical approaches in addition to visualization of calcium gradients in addition to oscillations. Using Arabidopsis as a model system, we provide the first genetic evidence to support a model in which calcium signals are natural regulators of pollen tube growth in addition to fertilization. Calcium signals are largely controlled by influx (through channels) in addition to efflux (through pumps in addition to antiporters). We have identified T-DNA gene disruptions in all 14 calcium pumps, in addition to all 20 cyclic nucleotide gated channels (CNGCs). From this set of mutants, distinct pollen specific phenotypes have been found as long as disruptions of ACA9 (a calmodulin activated plasma membrane calcium pump) in addition to CNGC18 (a cyclic nucleotide in addition to calmodulin regulated putative calcium channel). Calcium Pump Mutation aca9 Reduces Pollen Tube Growth Potential (in vivo) cngc18 -/+ have full seed set indicating that the defect is very different than the aca9 phenotype. Reciprocal crosses in addition to out-crossing to the male sterile mutant, ms-1 show that cngc18 is male sterile. CONCLUSIONS: funiculus funiculus aca9 disruption results in partial male sterility WT aca9 Longest pollen tube Aniline blue stain WT ACA9promoter::GUS ACA9 appears to function as a plasma membrane pump in pollen aca9 disruption results in >50% reduced frequency of synergid penetration (discharge) required as long as fertilization aca9 -/- blocked blocked fertilized Synergid penetration penetration / discharge Confocal image of GFP merged with DIC image No synergid pentration or sperm release Discharge + Discharge – Discharge +: Wild type 96% (n=178) aca9 41% (n=247) CNGC18 disruption results in complete male sterility No homozygous cngc18 in over 400 F1 progeny of two independent gene disruption lines. Homozygous gene disruption lines in all CNGCs but CNGC18. Alex in addition to er staining as long as pollen viability in the quartet background (qrt) shows that cngc18 pollen is normal developed in addition to viable. Expression profile of CNGCs during pollen development Modified after Maeser et al., 2001 CNGC8 in addition to CNGC18 are highly expressed in mature pollen. Only disruption of CNGC18 results in male sterility. 100x cngc18 pollen tubes germinate, but only grow a short distance, with a “kinky-like” non-directional growth, often prematurely terminating with a bursting event. cngc18 pollen germinate but has impaired tube growth in vitro 400x 400x control, qrt (-/-) cngc18 (-/+), qrt (-/-) In vitro germination assays in the quartet background (pollen doesn’t separate during development). control, qrt (-/-) cngc18 (-/+), qrt (-/-) aca9 is the first mutant with a defect in pollen/ovule interaction. CNGC18 is the only CNGC essential as long as the plant life cycle. CNGC18 is the first potential calcium channel that was found to be involved in pollen tube growth. ACA9 in addition to CNGC18 could be involve in a CaM regulated calcium oscillator, that is essential as long as pollen tube growth in addition to fertilization. aca9 -/- plants display reduced pollen tube growth in addition to >50% reduction of synergid penetration (discharge). ACA9: CNGC18: Model: cngc18 pollen germinates with underdeveloped, kinky pollen tubes, that often burst at the tip. Disruption of ACA9 results in partial male sterile plants with shorter siliques in addition to reduced seed set. Disruption of CNGC18 results in complete male sterility. Honys in addition to Twell, 2003, Extracted from NASC database WT,col cngc18 (-/+)

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