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The anther regeneration system is the earliest in vitro tissue culture regeneration plant system of Hevea brasiliensis established in China, and the regenerated plants were named self-rooted juvenile clone, which have the advantages of rapid growth, high resistance as seedlings and genetic homogeneity as grafted seedlings, and yield increase by more than 20% due to rejuvenation. However, the buds collection period is only about 60 d per year, which does not allow for a year-round supply of callus formed by dedifferentiation anther wall cells, resulting in unsustainable research relying on this material. In this study, Reyan73397 stamens were utilized to study the callus induction and somatic embryogenesis of anther callus at different developmental periods (0 d, 7 d, 14 d, 21 d, 28 d, 35 d and 42 d), which were stored at two low temperatures of 15 ℃ and 20 ℃ for 15 d. The results showed that temperature 15 ℃ and 20 ℃ had no significant effect on the stages of callus induction and somatic embryogenesis after preservation, except that the number of embryogenic callus was significantly lower than that of the control the developmental period had significant effect on callus induction and somatic embryogenesis after preservation, and showed the greatest effect on the stage of cotyledonary embryo formation, of which the anther callus at 21 d and 35 d responded strongly, the number of cotyledonary embryos of 21 d anther callus was significantly higher than that of the control, and the number of cotyledonary embryos of 35 d anther callus was significantly lower than that of the control after preservation; the preservation of anther callus with different developmental periods for 15 d at 15 ℃ and 20 ℃ could extend the supply time of anther callus for 15~60 d, and primary cotyledonary embryos for 27-95 d compared to the control; the optimal solution was to keep the anther callus with developmental periods at 21 d at 15 ℃ and 20 ℃ for 15days, which could prolong the supply of anther callus and primary cotyledonary embryo for 60 days and 60-95 days separately. Although the treatment reduced the number of callus by 46.61%, it promoted the formation of cotyledon embryo by 27.18% in number. In this study, the short-term preservation of H. brasiliensis anther callus was realized, which would greatly alleviate the difficulty of insufficient source of genetic transformation receptors due to the short flowering period, the study time could be carried out for the whole year from 60 days to 240 days, which could help to speed up the research on optimization of the genetic transformation system of anther callus in Hevea brasiliensis.
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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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