大花金挖耳无菌外植体的获得及愈伤组织诱导的初步研究

以大花金挖耳(C arp esium m acrocepha lum F ranch.et Sav)种子为材料,进行了种子消毒和愈伤组织诱导及继代的初步研究。结果表明,对大花金挖耳种子进行消毒的最佳条件为,以体积分数70%的乙醇消毒5 m in后,再用50%的N aC lO3处理30 m in。在大花金挖耳无菌苗的根形成愈伤组织的正交试验中,基本培养基对愈伤组织诱导率的影响最大,NAA次之,6-BA影响最小,对愈伤组织诱导的较优组合是B5+NAA 3.0 m g/L+6-BA 0.2m g/L,诱导率为100%;在优化诱导愈伤组织的激素浓度配比试验中,B5+NAA 2.0~3.0 m g/L+6-BA 0.2~0.4m g/L对大花金挖耳愈伤组织的诱导率为100%,且产生的愈伤组织多为淡黄色、质地疏松、生长势极好、褐化时间长,适合于大花金挖耳愈伤组织的诱导。在培养基中分别添加500 m g/L VC和10 000 m g/L PVP能抑制大花金挖耳愈伤组织的褐变,且有利于愈伤组织的继代。

Study on the obtaining of sterile explants and callus induction of Carpesium macrocephalum Franch.et Sav

It was the first time to induce the callus of Carpesiurn macrocephalurn Franch. et Sav by taking its root as the explants from its seeds in this paper. The results indicated that the asepsis germination rate of seed amounted to 98.80% when its seeds were sterilized with 70% alcohol for 5 min,then 50% NaClO3 for 30 minutes. It also showed that such factors as basic media, types and concentration of hormones had a great effect on the induction rate,growth and browning time of the callus. The proper callus induction media were selected by orthogonal design,more-over,the kind and suitable level of hormone were ascertained. The basic media were the critical factors to callus induction,then NAA,the optimum medium for callus induction of Carpesiurn macrocephalum Franch. et Sav was B5 4- NAA 3. 0 mg/L 4- 6-BA 0. 2 mg/L. Moreover the best effects were obtained by using NAA of 2.0-3.0 mg/L4-6-BA 0. 2--0. 4 mg/L in B5 solid mediums. The induction rate of the callus was 100%,and the callus was primrose yellow and loose. By adding 500 mg/L Vc or 10 000 mg/L PVP,the callus kept primrose yellow and vigorous.