摘 要:为分析十字花科植物体细胞胚胎的诱导发生规律,本文以蔊菜为实验材料,对蔊菜不同外植体进行了多种组合诱导体胚发生。在含有山梨醇、甘露醇和NaCl的胁迫培养基上诱导出“类蛙卵胚”结构,然后采用双染色法对其进行胚性鉴定;通过冷冻切片技术对体胚进行发育细胞形态学的检测与起源分析。实验结果表明,对蔊菜外植体(根)进行胁迫处理7-10天,显出愈伤结构,18-22天后,出现类蛙卵胚;双染色法鉴定的类蛙卵胚结构为鲜亮的红色,愈伤为深蓝色;冷冻切片分析该体胚起源于根的内皮层。本研究对于十字花科体胚发生诱导具有重要的理论价值。
关键词:体细胞胚胎;蔊菜;类蛙卵胚7735
The Studies on Somatic Embryogenesis of Rorippa indica
Abstract: In this paper, the Rorippa indica was made as the experimental material for the somatic embryogenesis through a variety of combinations with different explants, for the analysis of induction regularity on cruciferous plants somatic embryos. The frog egg-like body(FELB) structure was inducted on the stress medium supplemented with sorbitol, mannitol and NaCl, then the structure and identification of embryos were analyzed by double staining. And the frozen section technique was used on detection the cell morphology and origin analysis of embryonic development. The experimental results show that the callus structures were inducted after the Rorippa indica explants (root) on stress treatments for 7-10 days, 18-22 days later, the FELB appeared. And the double staining made the FELB to bright red, then callus were dark blue, the frozen section analysis of FELB from the endodermis of root explant. The studies has great theoretical value on cruciferous somatic embryogenesis induction.
Key words: Somatic embryos; Rorippa indica; Frog egg-like body
目 录
摘要 1
引言 1
1. 材料与方法 4
1.1 材料与仪器 4
1.1.1 材料 4
1.1.2 试剂 4
1.1.3 仪器设备 4
1.2 试验流程 4
1.3 实验方案 4
1.3.1 播种 4
1.3.2 取材 5
1.3.3 暗处理 5
1.3.4 光下培养 5
1.4 鉴定方法 5
1.4.1 宏观判断 5
1.4.2 显微镜分析 5
1.4.3 双染色法鉴定 5
1.4.4 光学与荧光显微镜分析 6
1.4.5 冷冻切片技术 6
2. 结果与分析 6
2.1 蔊菜体胚发生 6
2.2 胚性鉴定 7
2.3 体胚起源分析 8
3. 结论 8
参考文献 10
致谢 15
蔊菜体胚发生研究引言
蔊菜(Rorippa indica (L.) Hiern),蔊菜别名野油菜、塘葛菜、山芥菜、地豇豆、江剪刀草等,一年生或二年生草本。植株较粗壮,高20-50cm,无毛或具疏毛。茎单一或分枝,直立斜升。叶形多变化,基生叶和茎下部叶具长柄;叶片通常大头羽状分裂,长4-10cm,宽1.5-2cm,顶裂片大,边缘具不规则牙齿,侧裂片1-3对,上部叶片宽披针形或匙形,具短柄或耳状抱茎,边缘具疏齿。总状花序顶生或侧生,开花时花序轴逐渐向上延伸,花小,多数;萼片4片,直立,浅黄色而微带黄绿色,光滑无毛,灾区披针形或卵状长圆形,先端内凹;花瓣4瓣,鲜黄色,宽匙形或长倒卵形,长2.5-4mm,全缘,基部具有短而细的爪;雄蕊6个,4长2短,长雄蕊长约3mm,短雄蕊长约2.5mm;雌蕊1个,子房圆柱形,长2.5-3mm,花柱短粗,柱头略膨大,顶部扁平线状圆柱形,较粗壮,长1-2cm,直立或稍弯曲,成熟时果瓣隆起。种子每室2行,多数,淡褐色,宽椭圆形,近三角形或不规多角形,长0.5-0.7mm,表面有凹陷的大网纹。花期4-5月(8-9月偶有),花后果实渐次成熟。无瓣蔊菜,一年生或二年生草本,高10-30cm。植株光滑无毛,较柔弱。茎直立或者呈铺散状分枝。叶抽薄;基生叶及茎下部叶具柄,叶柄长3-5cm,两侧具窄翼;叶片常为大头羽状分裂,长柄、叶柄长3-5cm,顶裂片大,宽卵形或长椭圆形边缘具大小不整齐钝锯齿,侧裂片1-3对,宽披针形或长椭圆形,上至下渐小;茎上部宽披针形或长卵形,长36cm,宽1.5-2.5cm,边缘具不整齐锯齿或者近全缘,基部具短柄或无柄。总状花序顶生或生,开花后,花序渐延长;花无瓣或偶有退化的花瓣;萼片4,淡黄绿色,披针形,先端微带紫色并内凹,长2.5-3mm,雄蕊6,4长2短,长雄蕊长约3mm,短雄蕊长约2.5mm;雌蕊1,子房圆柱形,花柱不明显,柱头盘状。长角果细圆柱形,细长而直,长1.5-3cm;果瓣近扁平,光滑或稀有柔毛。种子每室1行,淡褐色,细小,多数,宽椭圆形、长方形或方形或不规则多角形。表面有小疣点及细网纹。花期4-9月,果期5-10月。叶片卵形或大头状羽裂。长角果细圆柱形或线形,长2cm以上,宽1mm。 蔊菜体胚发生研究+文献综述:http://www.751com.cn/shengwu/lunwen_5805.html