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The axillary buds, which are embedded in the stem of Phalaenopsis and lurked at the base of leaves, can germinate and elongate to form pedicels under suitable conditions. The microscopic observation of the differentiation process is one of the important research contents to understand the flower development process and influencing factors of Phalaenopsis. In order to establish a paraffin section method for two different tissues with soft and hard connection, the stem segment with axillary bud was used as the test material. The preparation steps, such as fixed time, softening liquid concentration and softening time, dehydration, etc., which mainly affect the paraffin section effect, were optimized. The results showed that vacuuming the fixative and extending the fixative time to 48 h or more were conducive to the full penetration of the fixative, and the fixative effect was the best. Using 10%, 12% hydrofluoric acid solution to soften for 12, 15 days and vacuuming for 30 min during softening could greatly reduce the brittleness and hardness of stem tissues with high lignification degree, meet the soft and hard requirements of sections, and effectively solve the problem of tissue separation and fragmentation caused by uneven softening. Dehydration treatment was set up with 7 step by step dehydration gradients from 30% to 100% ethanol, and the gradual replacement steps of anhydrous ethanol to xylene were added, which could avoid the shrinkage and deformation of tissue materials of Phalaenopsis and ensure the integrity of tissue morphology. The optimized preparation method avoids the problems such as incomplete softening, separation of tissues from paraffin wax, crumpling and crushing of wax bands, and cavity of sections caused by conventional preparation of the stem segment of the axillary bud of Phalaenopsis girdle, etc. The overall structure of the obtained sections tends to be complete, the staining is uniform and bright, and the tissue cells in each part are obviously colored. Young bud tissue and lignified stem tissue can be clearly distinguished. In this study, a set of paraffin section preparation method was developed which could be applied to the stem segment of Phalaenopsis axillary bud, a soft and hard tissue material, and would provide a technical basis for further study on the differentiation process of latent axillary bud germination of Phalaenopsis.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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