为了得到生长状态良好的胚性愈伤组织（EC）,本研究以红花玉兰的未成熟种子作为外植体,通过1/2 MS基本培养基添加不同浓度的2,4-D、6-BA、水解干酪素（CH）以及提高蔗糖浓度和使用脱落酸（ABA）两步法的措施来诱导胚性愈伤组织更好的生长,并通过测定其可溶性蛋白、可溶性糖以及游离脯氨酸等指标来进行差异分析。结果表明：最适合诱导胚性愈伤组织的培养基为1/2 MS＋2.0 mg/L 2,4-D＋0.2 mg/L 6-BA＋750 mg/L CH（诱导率高达96.8%）;蔗糖浓度提高到5%和6%时,胚性愈伤组织的状态较好,诱导率可达50.0%和64.7%,且测得胚性愈伤组织的可溶性蛋白和游离脯氨酸含量均显著高于其他处理,当蔗糖浓度为6%时,胚性愈伤组织的可溶性糖含量显著高于其他处理。结果表明胚性愈伤组织较非胚性愈伤（NEC）具有较高的代谢活性,且经高浓度蔗糖处理后的胚性愈伤对这一刺激作出了积极的响应,为体胚分化提供了科学的理论基础。
In order to obtain embryogenic callus with good growth status, the embryogenic callus（EC） were inducted from immature seeds of Magnolia wufengensis which were used as the explant. The seeds were cultured in 1/2 MS minimal medium with 2,4-D, 6-BA and casein hydrolysate（CH） in different concentrations, and then sucrose concentration was improved and abscisic acid（ABA） was added for better growth. Besides, soluble protein, soluble sugar and free proline of embryogenic callus were determined for differential analysis. The results showed that the content of the optimal induction medium was 1/2 MS, 2.0 mg/L 2,4-D, 0.2 mg/L 6-BA and 750 mg/L CH（induction rate up to 96.8%）. Embryogenic callus grew well when they were transferred to medium with sucrose concentration increasing to 5% and 6% and the induction rate could reach 50.0% and 64.7%, respectively.Meanwhile, the soluble protein and free proline content of embryogenic callus were significantly higher than other treatments. Especially when the sugar concentration was 6%, the soluble sugar content was significantly higher than other treatments. This phenomenon indicated that embryogenic callus had higher metabolic activity than non-embryogenic callus（NEC）. The embryogenic callus treated with higher concentration of sucrose had a positive response to this stimulus, which could provide a scientific basis for the differentiation of somatic embryos.
Molecular Plant Breeding
Physiology and biochemistry