以11份不同基因型胡萝卜为试材进行游离小孢子培养研究,并对胚状体及愈伤组织形成过程进行细胞学观察。诱导培养92d后,有5份材料形成肉眼可见的胚状体或愈伤组织。其中70198和80Q54均诱导出胚状体和愈伤组织,胚状体产率分别为每皿10.56和7.00个,愈伤组织产率分别为每皿3.56和8.38个;90234仅形成胚状体,产率为每皿6.75个;70Q78和80Q52仅产生愈伤组织,产率分别为每皿3.00和1.00个。细胞学观察表明胚状体和愈伤组织形成过程具有完全不同的结构特征。发育成胚状体的小孢子细胞壁变薄,膨大伸长,长度可至原来的1.5~4.0倍,有明显大液泡,初期细胞纵向分裂,成串排列,细胞之间紧密连接,而发育成愈伤组织的小孢子膨大成球状,多次分裂后松散地连接在一起。通过流式细胞仪鉴定70198、70Q78和80Q54获得的137株再生植株倍性,单倍体、二倍体及三倍体所占比例分别为68.6%、29.9%和1.5%。
The differences about the development of embryoids and calli arising from microspores were investigated in isolated microspore culture with eleven carrot accessions.After cultured about 92 days,five accessions began to form embryoids or calli.70198 and 80Q54 produced both,the productions of embryoids were 10.56 and 7.00 per dish,and the productions of calli were 3.56 and 8.38 per dish,respectively.90234 only produced embryoids,and the production was 6.75 per dish.70Q78 and 80Q52 only formed calli,and the productions were 3.00 and 1.00 per dish,respectively.Microspores with thin wall,size elongating 1.5-4.0 folds,and large vacuole would develop into embryoids.Firstly they latitude split,arranged in series and connected tightly each other.While the microspores expanding like globe shape,splitting and connecting loosely would grow up to calli.The ploidy level of 137 regenerated plants from 70198,70Q78 and 80Q54 were detected using flow cytometry,and 68.6% were haploid,29.9% double haploid,and 1.5% triploid.