A method for in vitro culture and rapid propagation of Centella asiatica was introduced. Stem segments with nodules were obtained from aseptic materials. Sterilized with 70% ethanol for 30 seconds, then soaked in mercury-raising solution of 1 g/L for 6 minutes, then rinsed with aseptic water for 5-8 times. The residual mercury-rising segments were rinsed clean and the adventitious buds were differentiated to intercept the noduled segments of aseptic seedlings, and transferred into MT+0.5 mg/LBA and MT+0.5 mg/LTDZ media. In the middle, the stem segments remain green. Adventitious buds grew from the stem nodes after 15 days. With the prolongation of culture time, the number of buds increased and cluster buds formed. However, the number of buds induced on MT + 0.5 mg/LTDZ medium increased significantly. MT+0.5mg/LTDZ was more conducive to the formation of cluster buds.
【技术实现步骤摘要】
一种积雪草离体培养和快速繁殖的方法
本专利技术涉及一种积雪草离体培养和快速繁殖的方法,具体地说是以一种积雪草离体培养和快速繁殖的方法。
技术介绍
目前公知的积雪草Centellaasiatica(L)Urb,又名崩大碗,为伞形科植物,以全草入药。积雪草性寒、味苦、辛,具清热利湿、解毒消肿的功效,临床上用于湿热黄疸、痈疮肿毒、跌打损伤、解砒霜中毒、覃中毒、解暑、传染性肝炎、流行性脑脊髓膜炎等。积雪草产于印度、日本、中国、印度尼西亚、南非、斯里兰卡等国,在我国主要分布于江苏、浙江、福建、江西、广东、广西等省区。积雪草作为草药使用在我国已有数千年历史,其有效组分积雪草总甙在国内外也广泛应用于临床。目前,积雪草主要依赖于野生资源,难以满足临床及中成药生产的需要,同时,大量采挖野生药材,造成资源枯竭及生态环境的恶化。
技术实现思路
材料1植物材料积雪草采自广西野生资源,经广州中医药大学药用植物教研室潘超美教授鉴定为伞形科植物积雪草Centellaasiatica(L.)Urb.,取其带节茎段,进行表面消毒和培养。离体培养根据试验要求,选用的基本培养基为MurashgeandTucker(MT),根据试验的不同阶段添加不同浓度的植物激素:不定芽分化试验添加6苄基腺嘌呤(6benzylaminopurine,BA)、N苯基N′1,2,3噻二唑5脲(thidiazuron,TDZ);根的诱导试验添加萘乙酸(naphthaleneacid,NAA)、吲哚丁酸(indolebutyricacid,IBA)及吲哚乙酸(indoleaceticacid,IAA)。每天光照12h,培养 ...
【技术保护点】
1.一种积雪草离体培养和快速繁殖的方法,无菌材料的获得取积雪草带节茎段,用体积分数为70%的乙醇浸泡30s,转入1g/L的升汞溶液中浸泡消毒6min,再用无菌水漂洗5~8次,把残留升汞漂洗干净,不定芽分化截取无菌苗的带节茎段,移入MT+0.5mg/LBA及MT+0.5mg/LTDZ的培养基中,茎段保持绿色;15d后从茎节处陆续长出不定芽,随着培养时间延长,芽的数量增多,形成丛生芽;而在MT+0.5mg/LTDZ培养基上诱导出的芽明显增多;MT+0.5mg/LTDZ更有利于丛生芽的产生。
【技术特征摘要】
1.一种积雪草离体培养和快速繁殖的方法,无菌材料的获得取积雪草带节茎段,用体积分数为70%的乙醇浸泡30s,转入1g/L的升汞溶液中浸泡消毒6min,再用无菌水漂洗5~8次,把残留升汞漂洗干净,不定芽分化截取无菌苗的带节茎段,移入MT+0.5mg/LBA及MT+0.5mg/LTDZ的培养基中,茎段保持绿色;15d后从茎节处陆续长出不定芽,随着培养时间延长,芽的数量增多,形成丛生芽;而在MT+0.5mg/LTDZ培养基上诱导出的芽明显增多;MT+0.5mg/LTDZ更有利于丛生芽的产生。2.根据权利要求1所述的方法,其特...
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