仙茅叶片的组织培养及其细胞学观察

Tissue culture and cytological observations of leaf explants of Curculigo orchioides

目的通过对仙茅叶片组织培养的研究与细胞学观察,为仙茅的快速繁殖提供技术依据。方法将仙茅幼叶培养在MS培养基,通过设计不同的光照条件,附加不同的激素、水解酪蛋白和活性炭成分,调查出愈率和成苗率。细胞学观察采用石蜡切片法。结果对仙茅叶片的愈伤诱导,黑暗的效果好于光照;在所试验的培养基成分中,适宜的激素配比是2.0mg/L2,4-D或6-BA1.5mg/L+2,4-D2.5mg/L,并且附加300mg/L水解酪蛋白和0.2%活性炭,对于仙茅叶片的离体成苗较好;培养后,愈伤组织主要由叶片的中脉产生,位于中脉上表皮内侧的薄壁细胞首先启动分裂,随后维管束鞘薄壁细胞及其外侧的叶肉细胞也启动分裂,参与愈伤组织的形成。愈伤分化时,芽发生于愈伤组织的表面,根发生于愈伤组织的内部。结论可以通过仙茅幼叶的组织培养进行仙茅的快速繁殖。

Objective The studies on tissue culture and cytological observations of leaf explants of Curculigo orchioides were conducted in order to provide the basis for the rapid propagation of C. orchioides. Methods Young leaf explants of C. orchioides were cultured on MS basal media. Differences in the callus induction and plantlet regeneration rate were observed by different light treatment as well as chemical factors like different phytohormones, casein hydrolysate （CH）, and activated charcoal （AC） concentrations. Paraffin method was used to cytological observation. Results For callus induction of leaf explants of C. orchioides, dark treatment gave better results compared to light treatment ; among the media tested, the suitable phytohormone combinations were 2.0 mg/L 2, 4-D or 6-BA 1.5 mg/L＋2, 4-D 2.5 mg/L, and 300 mg/L CH＋0.2% AC was good for plantlet regeneration from leaf explants. The callus from leaf explants mainly originated from midrib. The parenchyma cells near epicuticle of midrib firstly were initiated to division. Then the parenchyma cells of vascular bundle sheath and mesophyll cells on each side of vascular bundle were also divided to form callus. The buds developed on the peripheral parts of the calli, but the roots developed in the regions deep within the calli. Conclusion Tissue culture of young leaf explants of C. orchioides can make the propagation of C. orchioides rapid.