黑麦草幼穗离体培养及植株再生

In vitro culture and plant regeneration from immature ears of Lolium perenne and Lolium multiflorum varieties

以多年生黑麦草和一年生黑麦草的适宜发育期(1～3mm)的幼穗为外植体,在附加适宜浓度2,4-D(1mg/L)的改良MS培养基上诱导愈伤组织发生,诱导率可达95%。一些愈伤组织在诱导培养基上直接发生不定芽,另一些愈伤组织能多次继代培养并保持高分化能力。后类愈伤组织在转入含有0～0.5mg/L6-BA和0～0.5mg/L2,4-D的分化培养基后产生大量丛生芽,部分小芽发育成苗。幼穗发育时期、培养基的激素组成明显影响愈伤组织的继代培养和植株再生能力,基因型对愈伤组织诱导率和植株再生能力也有影响。该体系适用于黑麦草基因工程和细胞工程研究,且具有实验周期短、基因型制约小、植株再生率高等优点。

Calli were induced from segments of immature ears of Lolium perenne and Lolium multiflorum varieties respectively on modified MS media supplemented with appropriate concentration of 2,4-D, usually 1 ((mg/L)). The frequency of calli induction could reach 95%. Some calli developed adventitious buds directly on the induction media. Some calli with higher ability of differentiation produced large amounts of buds or shoots after being cultured on the differentiation media. The factors influenced the frequency of calli induction and plantlet regeneration were evaluated. The development stage of the immature ears and the combination of 6-BA and (2,4-D) in the media played important roles on the calli subculture and plant regeneration as well as the genotypes of ryegrass affected the calli induction and plant regeneration. This system, with advantages of short-period, less genotype-dependence and higher plantlet regeneration, was available for cell engineering and (genetic) manipulation for ryegrass.