香根草组织培养技术的研究

香根草常规的无性繁殖方式难以满足运用生物技术筛选香根草抗性种质对种苗的需求,更难以满足大规模的香根草生态工程对种苗的需求。因此,创建香根草的组织培养技术是很有必要的。以香根草的腋芽为外植体进行离体培养,以MS为基本培养基,对胚性愈伤组织的诱导和植株再生体系的建立进行了优化。结果表明,适宜的诱导愈伤培养基为MS 2.0 mg/L生长素(2,4-D) 1.0 mg/L细胞分裂素(6-BA),愈伤组织诱导率最高可达96.7%;适宜的分化培养基为MS 1.0 mg/L 6-BA;适宜的生根培养基为1/2 MS 0.1 mg/L吲哚丁酸(IBA) 0.1 mg/L多效唑(PP333)。不同品种在相同培养条件下的愈伤诱导率存在较大差异。香根草的胚性愈伤组织具有单子叶植物典型的胚胎结构,其植株再生能力在继代条件下可以长期保持,继代18次的愈伤组织植株再生能力仍高达92.0%,即使继代24次后仍可达89.6%。香根草高效再生系统的建立为香根草开展基因工程及遗传转化方面的研究奠定了基础,也为大规模繁殖香根草种苗提供了新的技术保障。

Study on tissue culture of Vetiveria zizanioides

Conventional asexual propagation methods for vetiver seedlings have not matched up with the huge demands of Vetiveria zizanioides eco-engineering,bio-technology and genetic engineering of V.zizanioides.Therefore,it is necessary to establish tissue culture techniques that are highly effectively for multiplying V.zizanioides.Axillary buds of V.zizanioides were used as explants to culture in vitro on Murashige and Skoog(MS) basal culture medium.The best reducing medium was MS 2.0 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D) 1.0 mg/L 6-benzyladenine(6-BA),and the highest induction frequency was up to 96.7%.The best differentiation medium was MS 1.0 mg/L 6-BA,and the best rooting medium was 1/2 MS 0.1 mg/L indole-3-butyric acid(IBA) 0.1 mg/L poclobutrazol(PP333).V.zizanioides callus has the typical monocotyledon embryo structure.The regeneration ability of embryogenic calli could be preserved and reached 92.0% after subculturing 18 times.It was still up to 81.6% after 24 subcultures.The establishment of an effective regeneration system of V.zizanioides will be valuable for future research in gene engineering and genetic transformation of V.zizanioides.