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本研究旨在离体培养条件下建立起海雀稗植株再生的技术体系,为开展海雀稗细胞和分子生物学育种工作创造条件。以Adalay海雀稗(Paspalum vaginatumSw.cv.Adalay)的幼穗为外植体材料,在附加2,4-D 2.0~4.0 mg/L稍作修改的MS培养基上,愈伤组织诱导率达90%以上。不同发育时期的幼穗影响其愈伤组织的诱导发生频率:1.0~2.0 cm长的幼穗,愈伤组织诱导率达80%以上;2.0 cm以上的幼穗,诱导率降至60%以下。在添加2,4-D2.0 mg/L和BAP 0.05 mg/L的愈伤组织诱导培养基上,诱导产生的颗粒状愈伤组织占愈伤组织总数的40%以上。愈伤组织继代培养基中琼脂的用量提高到16 g/L时,愈伤组织在继代培养过程中保持颗粒状结构。颗粒状愈伤组织经连续继代培养3次后转移到附加BAP 2.0 mg/L的分化培养基上,植株再生频率达98%。通过提高愈伤组织继代培养基的渗透压,海雀稗幼穗诱导产生的颗粒状愈伤组织在继代培养过程中能够保持颗粒状的结构和高频率植株再生的能力。
The aim of this study is to establish a technical system for the regeneration of A. paspalum plantlets in vitro and to create the conditions for the breeding of B. barnyardis cells and molecular biology. Using the young spike of Adalay paspalum (Paspalum vaginatumSw.cv.Adalay) as explant material, the callus induction rate of MS medium supplemented with 2,4-D 2.0-4.0 mg / L More than 90. The young ear at different developmental stages affected the induction frequency of callus: the young callus with the length of 1.0 ~ 2.0 cm, the callus induction rate was over 80%; the young callus above 2.0 cm reduced the callus induction below 60%. Callus induction induced by adding 2,4-D2.0 mg / L and BAP 0.05 mg / L accounted for more than 40% of the total number of callus. When the amount of agar in the callus subculture medium increased to 16 g / L, the callus maintained the granular structure during the subculture. The granular callus was subcultured three times in succession and then transferred to a differentiation medium supplemented with 2.0 mg / L BAP. The frequency of plant regeneration reached 98%. By increasing the osmotic pressure of callus subculturing medium, the granular callus induced by young sparrow panicle can maintain the granular structure and the ability of high-frequency plant regeneration during subculture.