摘要
大麦小孢子单核中期的穗,经4℃预处理4星期后,直接剪成小段,捣碎、过滤、离心和洗涤后收集小孢子。用MSm+Maltose60.0g/L+1.0mg/LBAP对小孢子慢速振荡培养,7天后可观察到小孢子分裂的小细胞团,经3星期左右培养,小细胞团总数达最高,其频率为总小孢子的2.36%。这些小细胞团80%发育为类似体细胞胚结构,20%为典型愈伤组织,类似体细胞结构胚置于1/2MS+1.0mg/LBAP+0.2mg/LNAA+0.2mg/LZT分化培养基上,总分化频率为67%,其中绿苗率为39%,愈伤组织经胚性改造后,在1/2MS+4.0mg/LBAP+0.5mg/LNAA+0.5mg/LIAA培养基上分化,总分化频率为62%,绿苗率为18%。
Microspores were isolated from the spike of the six-row barley (Hordeum vulgare L,) cultivarFrances at the mid-unicellular pollen stage, pretreated at 4℃ for 4 weeks, by directly cutting the spike intosmall segments and homogenizing. The isolated microspores were incubated in MSm medium supplementedwith 60 g/ L maltose, 1.0mg/ L BAP with slowly shaking at 25℃ in darkness. Cell aggregatesfrom microspores were observed after 7 days from in initial culturing and the number of them reached tothe highest point at a frequency of 2.36% after about 3 weeks of culture. The cell aggregates developedembryo-like structure and call us at the rate of 80% and 20%, respectively. The embryo-like structureson 1/2 MS containing 1.0mg/ L BAP, 0.2mg/ L NAA and 0.2mg/ L ZT regenerated green plantlets(39%) and albino plantlets (28%). The calli by being improved on L3 medium with 2.0 mg / L 2, 4-D,0.1mg / L ABA and 0.2mg / L ZT, regenerated green plantlets (18%) and albino plantlets (44%), on 1 / 2MS medium by adding 4.0 mg / L BAP, 0.5mg / L N AA and 0.5mg/ L 1AA.
出处
《浙江农业学报》
CSCD
1995年第3期182-186,共5页
Acta Agriculturae Zhejiangensis
关键词
六棱大麦
游离小孢子
细胞团
植株再生
大麦
Six-row barley (Hordeum vulgare L.)
Isolated microspore
Cell aggregate, Plant regeneration