Figure S3.
Two pollen ADF isovariants from S. lycopersicum show pH-dependent activity in the in vitro actin filament disassembling assay. (A) Phylogenetic analysis of ADFs from several plant species. The maximum-likelihood phylogenetic tree of ADFs from several plant species was generated by the Maximum Likelihood method based on the JTT matrix-based model (Jones et al., 1992). Four subclasses of ADFs are highlighted with different colors. Species abbreviations are as follows: At, Arabidopsis thaliana; Zm, Zea mays; Os, Oryza sativa; Solyc, S. lycopersicum; Bd, Brachypodium distachyon; Php, Physcomitrella patens. PhpADF was used as an outlier. The indicated numbers of the branch nodes in the phylogenetic tree refer to posterior probabilities. Bootstrap = 500. The protein accession numbers from GeneBank were as follows: AtADF1, NP_190187; AtADF2, NP_566882; AtADF3, NP_851227; AtADF4, NP_851228; AtADF5, NP_565390; AtADF6, NP_565719; AtADF7, NP_194289; AtADF8, NP_567182; AtADF9, NP_195223; AtADF10, NP_568769; AtADF11, NP_171680; Solyc01g094400, XP_019068822; Solyc01g111380, XP_004231029; Solyc03g025750, XP_010317604; Solyc04g011370, XP_004236758; Solyc06g005360, XP_004240356; Solyc06g035980, XP_004240732; Solyc09g010440, XP_004246462; Solyc09g072590, XP_004247519; Solyc09g090110, XP_004247731; Solyc10g017550, XP_019071428; Solyc10g084660, XP_004249528; OsADF1, XP_015622816; OsADF2, XP_015630947; OsADF3, XP_015632173; OsADF4, XP_015632174; OsADF5, XP_015631622; OsADF6, XP_015637229; OsADF8, XP_015644676; OsADF9, XP_015645783; OsADF10, XP_015613388; OsADF11, XP_015618885; Zm2G015127, NP_001148445; ZmADF1, XP_008651860; ZmADF2, NP_001105590; ZmADF3, CAA66311; ZmADF5, XP_008681563; ZMADF6, ACG32422; BdADF1, XP_003570044; BdADF2, XP_003558421; BdADF3, XP_010227648; BdADF4, XP_010227656; BdADF5, XP_003558363; BdADF6, XP_014751334; BdADF9, XP_010238336; BdADF10, XP_003574212; PhpADF, PNR26890. (B) Tissue expression of subclass II ADFs from S. lycopersicum. Based on the phylogenetic analysis shown in A, four S. lycopersicum ADFs (Solyc01g094400 (SlADF1), Solyc03g025750 (SlADF3), Solyc06g035980 (SlADF6), and Solyc10g017550 (SlADF10)) were classified into subclass II. The original tissue expression datasets were retrieved from the eFP Browser (http://bar.utoronto.ca/). (C) The expression of SlADF1, SlADF3, and SlADF10 in S. lycopersicum pollen was assessed by RT-PCR analysis. (D and E) SDS-PAGE analysis of samples from high-speed F-actin cosedimentation experiments. [F-actin], 3 μM; [ADF], 20 μM. S, supernatant; P, pellet. (F) The amount of actin in the supernatant was quantified and plotted versus pH. Data are presented as mean ± SD, n = 3. Source data are available for this figure: SourceData FS3.

Two pollen ADF isovariants from S. lycopersicum show pH-dependent activity in the in vitro actin filament disassembling assay. (A) Phylogenetic analysis of ADFs from several plant species. The maximum-likelihood phylogenetic tree of ADFs from several plant species was generated by the Maximum Likelihood method based on the JTT matrix-based model (Jones et al., 1992). Four subclasses of ADFs are highlighted with different colors. Species abbreviations are as follows: At, Arabidopsis thaliana; Zm, Zea mays; Os, Oryza sativa; Solyc, S. lycopersicum; Bd, Brachypodium distachyon; Php, Physcomitrella patens. PhpADF was used as an outlier. The indicated numbers of the branch nodes in the phylogenetic tree refer to posterior probabilities. Bootstrap = 500. The protein accession numbers from GeneBank were as follows: AtADF1, NP_190187; AtADF2, NP_566882; AtADF3, NP_851227; AtADF4, NP_851228; AtADF5, NP_565390; AtADF6, NP_565719; AtADF7, NP_194289; AtADF8, NP_567182; AtADF9, NP_195223; AtADF10, NP_568769; AtADF11, NP_171680; Solyc01g094400, XP_019068822; Solyc01g111380, XP_004231029; Solyc03g025750, XP_010317604; Solyc04g011370, XP_004236758; Solyc06g005360, XP_004240356; Solyc06g035980, XP_004240732; Solyc09g010440, XP_004246462; Solyc09g072590, XP_004247519; Solyc09g090110, XP_004247731; Solyc10g017550, XP_019071428; Solyc10g084660, XP_004249528; OsADF1, XP_015622816; OsADF2, XP_015630947; OsADF3, XP_015632173; OsADF4, XP_015632174; OsADF5, XP_015631622; OsADF6, XP_015637229; OsADF8, XP_015644676; OsADF9, XP_015645783; OsADF10, XP_015613388; OsADF11, XP_015618885; Zm2G015127, NP_001148445; ZmADF1, XP_008651860; ZmADF2, NP_001105590; ZmADF3, CAA66311; ZmADF5, XP_008681563; ZMADF6, ACG32422; BdADF1, XP_003570044; BdADF2, XP_003558421; BdADF3, XP_010227648; BdADF4, XP_010227656; BdADF5, XP_003558363; BdADF6, XP_014751334; BdADF9, XP_010238336; BdADF10, XP_003574212; PhpADF, PNR26890. (B) Tissue expression of subclass II ADFs from S. lycopersicum. Based on the phylogenetic analysis shown in A, four S. lycopersicum ADFs (Solyc01g094400 (SlADF1), Solyc03g025750 (SlADF3), Solyc06g035980 (SlADF6), and Solyc10g017550 (SlADF10)) were classified into subclass II. The original tissue expression datasets were retrieved from the eFP Browser (http://bar.utoronto.ca/). (C) The expression of SlADF1, SlADF3, and SlADF10 in S. lycopersicum pollen was assessed by RT-PCR analysis. (D and E) SDS-PAGE analysis of samples from high-speed F-actin cosedimentation experiments. [F-actin], 3 μM; [ADF], 20 μM. S, supernatant; P, pellet. (F) The amount of actin in the supernatant was quantified and plotted versus pH. Data are presented as mean ± SD, n = 3. Source data are available for this figure: SourceData FS3.

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