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To establish a regeneration system for somatic embryo-derived plants from the anthers of the triploid rubber tree clone Yunyan 73-46, this study utilized anthers as explants and employed an orthogonal experimental design to examine the effects of various hormone combinations on callus and somatic embryo induction efficiency. Furthermore, the morphological characteristics and differentiation capacity of callus under different culture duration were analyzed. Hormones were crucial for callus induction. In the absence of hormones, anthers failed to dedifferentiate and underwent browning and necrosis after 10d. In media containing 2,4-D, NAA, KT and TDZ, anther dedifferentiation to produce callus occurred with induction rates ranging from 15% to 91.56%, with a mean of (69.41±30.37)%. The impact of 2,4-D, NAA, KT and TDZ on callus induction rate and subsequent somatic embryogenesis rate were all highly significant (P<0.01), with the order of influence being 2,4-D>TDZ>NAA>KT. The optimal combination identified through comprehensive screening was 2,4-D 1.0 mg/L+NAA 0.8 mg/L+TDZ 0.02 mg/L. Upon verification, the callus induction rate and subsequent somatic embryogenesis rate was (86.15±1.53)% and (14.46±0.89)%, respectively, marking significant improvements of 2.31% and 5.40% over the best experimental combination (G7). The somatic embryogenesis capacity of this clone was generally low, with somatic embryo formation rates ranging from 6.91% to 19.31%, with a mean of (10.58±3.95)%. The effects of 6-BA, ABA, NAA and KT on somatic embryogenesis rate were highly significant (P<0.01), with the order of influence being ABA>6-BA>NAA>KT. The optimal combination was 6-BA 0.4 mg/L+ABA 0.2 mg/L+NAA 0.2 mg/L+KT 0.2 mg/L (M9), achieving a somatic embryogenesis rate of (19.31±0.60)%, which was significantly higher than other treatments. The optimal culture duration for callus was determined to be 55 d, which exhibited superior growth status without browning and the strongest somatic embryo differentiation capacity. The somatic embryogenesis rate was (30.83±1.67)%, the cotyledonary embryo formation rate was (18.33±0.83)%, and the average number of somatic embryos was (0.63±0.03), all of which were extremely significantly higher than those at other culture durations. By optimizing the hormone ratios during callus induction and somatic embryogenesis stages of Yunyan 73-46 anthers, coupled with adjusting the duration of callus induction culture, the production efficiency of anther-derived somatic embryo plantlets can be significantly enhanced. This approach would provide technical support for establishing a large-scale propagation system of somatic embryo plantlets for triploid rubber tree clones.
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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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