Bhattacharya Sabita, Bhattacharyya Sanghamitra
Department of Botany, Bose Institute, Kolkata, India.
Methods Mol Biol. 2010;589:117-26. doi: 10.1007/978-1-60327-114-1_12.
The growing demand for flower extracts in perfume trade can primarily be met by increasing flower production and multiplying planting material. The major commercial aromatic flower yielding plants including Jasminum officinale L., a member of the Family Oleaceae have drawn the attention of a large section of the concerned sectors leading to a thrust upon developing advanced propagation technologies for these floral crops, in addition to conventional nature-dependent agro-techniques. This chapter describes concisely and critically, a protocol developed for in vitro propagation of Jasminum officinale by shoot regeneration from existing as well as newly developed adventitious axillary buds via proper phytohormonal stimulation. To start with nodal segments as explants, March-April is the most ideal time of the year when planting material suitable for in vitro multiplication is abundantly available. Prior to inoculation of explants in the culture medium, special care is needed to reduce microbial contamination by spraying on selected spots of the donor plant with anti-microbial agents 24 h prior to collection; treatment with antiseptic solution after final cleaning and surface sterilization by treating explants with mercuric chloride. Inoculated explants are free from brown leaching from cut ends by two consecutive subcultures within 48 h in MS basal medium. Multiplication of shoots, average 4-5 at each node, takes place in MS medium containing 4.0 mg/L BAP, 0.1 mg/L NAA, and 40 g/L sucrose over a period of 8 weeks. For elongation of regenerated shoots, cultures are transferred to MS medium, supplemented with a single growth hormone, kinetin at 2.0 mg/L. Emergence and elongation of roots from shoot base is facilitated by placing on the notch of a filter paper bridge. The hardened in vitro propagated plants are able to grow normally in soil like other conventionally propagated Jasminum officinale.
香水行业对花卉提取物日益增长的需求，主要可以通过增加花卉产量和繁殖种植材料来满足。主要的商业芳香花卉生产植物，包括木犀科的素馨属植物素馨，已经引起了相关领域很大一部分人的关注，这使得除了传统的依赖自然的农业技术外，还大力推动为这些花卉作物开发先进的繁殖技术。本章简要且批判性地描述了一种通过适当的植物激素刺激，从现有的以及新发育的不定腋芽进行芽再生，从而实现素馨体外繁殖的方案。以节段作为外植体开始培养，3月至4月是一年中最理想的时间，此时有大量适合体外繁殖的种植材料。在将外植体接种到培养基之前，需要特别小心以减少微生物污染，即在采集前24小时用抗菌剂喷洒供体植物的选定部位；在最终清洁并用氯化汞处理外植体进行表面消毒后，再用防腐溶液处理。接种后的外植体在MS基本培养基中48小时内通过连续两次继代培养，切口端无褐色渗出物。在含有4.0mg/L BAP、0.1mg/L NAA和40g/L蔗糖的MS培养基中，经过8周时间，每个节点平均可产生4 - 5个芽进行增殖。为了使再生芽伸长，将培养物转移到添加了单一生长激素2.0mg/L激动素的MS培养基中。通过放置在滤纸桥的切口上，有助于从芽基部生出根并伸长。经过硬化处理的体外繁殖植物能够像其他传统繁殖的素馨一样在土壤中正常生长。
