剑麻愈伤组织的诱导和再生体系的建立

Establishment of Sisal Callus Induction and Shoot Regeneration System

以剑麻H.11648为材料,研究外植体类型、激素种类及其浓度对剑麻愈伤组织的诱导和植株再生的影响,并且建立剑麻茎尖愈伤组织诱导及高频再生体系。结果表明:MS基本培养基、6-BA/NAA激素组合有利于淡黄绿色愈伤组织的形成,SH基本培养基、6-BA/IBA/NAA激素组合有利于不定芽的分化;最佳愈伤组织诱导和分化培养基分别为MS+3.0 mg/L 6-BA+0.2 mg/L NAA,SH+5.0 mg/L 6-BA+0.1 mg/L NAA+0.1 mg/L IBA。在该培养条件下,H.11648麻茎尖愈伤组织的诱导率为86.67%,分化率为(98.33±1.67)%,分化系数为(13.19±0.58)。再生植株在不加激素的MS基本培养基上培养4周后的生根率为100%,平均生根数为(5.39±0.70)条,根长为(8.44±0.25)cm,该研究结果为剑麻转基因的研究和种质创新奠定了基础。

The effects of explant types, combination and concentration of plant growth regulators on callus induction and shoot regeneration in Agave hybrid cv No.11648 were examined. The results showed that MS basal medium supplied with 6-BA and NAA was suitable for the light-yellow green callus induction, SH basal medium supplied with 6-BA, NAA and IBA was suitable for adventitious differentiation. The highest rate of light yellow- green, compact and nodular-like calli was formed on the MS medium supplemented with 3.0 mg/L BA in combination with 0.2 mg/L NAA, with 86.67% of shoot tips producing a compact, light yellow-green and nodular- like callus. The calli derived from the different explants were subcuhured on Schenk and Hildebrandt （SH） medium containing different concentrations of 6-BA, NAA and indole-3-butyric acid（IBA）for shoot induction. Shoot organogenesis was achieved when the callus tissue derived from shoot tips was transferred to the SH medium. Maximum organogenesis was obtained with 5.0 mg/L 6-BA in combination with 0.1 mg/L NAA and 0.1 mg/L IBA, yielding （98.33 ±1.67）% shoot induction frequencies and （13.19±0.58）adventitious differentiation coefficient. The regenerated shoots were rooted on hormone-free MS medium, with an average number of 5.39 roots per shoot, and a length of 8.44 cm per root within 30 days, and acclimatized in a greenhouse, with more than 95% survival. These results establish foundation for transgenic technology and germplasm innovation in sisal.