2.1.2. Testing for autogamy (auto- or self-pollination)
This test will tell us whether the flower can set seeds and fruits from its own pollen. In such a situation, the contribution of flower visitors may be little or none, but even in auto-pollinating plants, the movements of bees inside the flower can sometimes optimize pollen transfer from anthers to the stigma and increase fruit- or seed-set. Auto-pollination or self-pollination should be distinguished from geitonogamy (see section 2.1.3.). Auto-pollination is associated with hermaphroditic flowers and pollen transfer within that flower that is automatic (soybean) or pollinator-optimized, whereas geitonogamy could apply to monoecious plants in which pollen is self-compatible but the actions of a pollen vector are nevertheless needed.
- Choose a given number of flower buds prior to anthesis. The investigator must become familiar with the flowering pattern of the model plant because flowers in some species open and close more than once, making anthesis difficult to determine. The number of buds may vary with availability and ease of access, but larger samples produce more reliable results.
- Protect two thirds of these buds with pollination bags (see section 2.1.1.) and leave the other third unbagged as open controls, or in the case of pollinator shortage pollinate these flowers manually with pollen from another plant of the same species. Identify each treatment with weather-resistant tags.
- After anther dehiscence and when stigmas are receptive, remove the bags of half of the protected flowers (one third of the total marked buds) and hand-pollinate the stigmas with a soft brush using pollen from the anthers of the same flower. Dehiscence can usually be recognized as anthers with a split in the anther wall, pore, or flap that is exposing the pollen. After hand-pollinating, re-bag the flowers to prevent flower visitors or wind pollination. Leave bags on flowers until they are no longer receptive, then remove the bags.
- At the end of the season, check whether fruit developed from the flowers that remained bagged throughout the experiment. If all or most of these flowers have developed into fruit, the plant species is autogamous and its flowers are capable of auto-pollinating. Honey bees can contribute little to increasing fruit- or seed-set. If only the hand-pollinated flowers developed into fruits, this means that the plant species is autogamous but flowers need a pollinating agent to transfer the pollen grains from their anthers to the stigmas within the flower. In this case, honey bees may be of great value. The proportion of fruit- or seed-set obtained from the bagged treatment in comparison to the hand-pollinated treatment will tell the comparative strength of autogamy in this plant species (strictly autogamous, highly autogamous, etc.). If no bagged flowers produce fruit or seeds, this means the species may be self-incompatible and probably needs cross pollen to set fruits and seeds. However, sometimes a few fruits or seeds can set even in self-incompatible plants because self-recognition can be incomplete. But in this case, there is little variation in fruit- or seed-set among the treated plants. One should not confound self-incompatibility with self-sterility resulting from inbreeding depression because in the latter case seed set varies greatly among treated flowers, ranging from low values in more inbred plants to high values in less inbred ones. Confirmation of self-incompatibility must be done by examining pollen tube growth in the pistil, a subject covered in section 3.1.5.