Table 2.
Purification of the Stylar Adhesive Molecule using an Adhesion Assay with SCA
| No. of Adhered Pollen Tubes
|
||
|---|---|---|
| Fractionsa | + SCA(SG)b | + SCA(SE)c |
| Imidazole extract | 227 ± 52 (6) | 328 ± 50 (3) |
| Ethanol precipitation of imidazole extract | ||
| Ethanol 20 to 40% insoluble | 77 ± 50 (2) | —d |
| Ethanol 40 to 60% insoluble | 236 ± 86 (8) | 334 ± 26 (3) |
| Ethanol 60 to 80% insoluble | 101 ± 30 (2) | — |
| Q Sepharose of ethanol 40 to 60% insoluble | ||
| Unbound | 35 ± 34 (3) | — |
| 100 mM NaCl eluted | 13 ± 13 (3) | — |
| 200 mM NaCl eluted | 10 ± 6 (3) | — |
| 300 mM NaCl eluted | 58 ± 40 (3) | — |
| 400 mM NaCl eluted | 220 ± 29 (4) | — |
| 500 mM NaCl eluted | 39 ± 24 (3) | — |
| 2 M NaCl eluted | 46 ± 26 (4) | — |
| Sepharose CL-4B of the 400 mM NaCl elutede | ||
| Fraction A | — | 320 ± 16 (2) |
| Fraction B | — | 261 ± 29 (2) |
| Fraction C | — | 401 ± 53 (5) |
| Fraction D | — | 204 ± 66 (2) |
| Fraction E | — | 286 ± 38 (2) |
| Fraction F | — | 20 ± 11 (2) |
| Fraction G | — | 298 ± 82 (2) |
| Fraction H | — | 37 ± 24 (2) |
| Fraction I | — | 13 ± 5 (2) |
| Fraction J | — | 1 ± 1 (2) |
None of the imidazole extracts (75 μg) noted in this table allowed pollen tube adhesion when used on their own (i.e., <20 adhered pollen tubes with at least three replicates for each extract).
Number of pollen tubes adhered to a nitrocellulose membrane coated with 75 μg of imidazole extract combined with 10 μg of SCA partially purified SCA(SG). The numbers in parentheses indicate the number of replicates.
Number of pollen tubes adhered to a nitrocellulose membrane coated with 50 μg of imidazole extract combined with 5 μg of SCA isolated from SCA(SE). The numbers in parentheses indicate the number of replicates.
(—), not determined.
Fractions of the size exclusion Sepharose CL-4B column are identified as shown in Figure 1.