Assessment of genetic homogeneity and analysis of phytomedicinal potential in micropropagated plants of Nardostachys jatamansi, a critically endangered, medicinal plant of alpine Himalayas
Assessment of genetic homogeneity and analysis of phytomedicinal potential in micropropagated plants of Nardostachys jatamansi, a critically endangered, medicinal plant of alpine Himalayas
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DOI：
10.1007/s11240-015-0897-x
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发表时间：
2016-02-01
影响因子：
3
通讯作者：
Tandon, Pramod
中科院分区：
文献类型：
--
作者：
Bose, Biswajit;Kumaria, Suman;Tandon, Pramod
甘松（Nardostachys jatamansi，D.唐）华盛顿，一种小型的多年生地下茎草本植物，自古以来就具有巨大的药用价值，仅限于海拔3000 - 5200米的喜马拉雅山脉的特殊生境。该物种最近已被列为严重濒危物种的IUCN红色名录，由于过度开发其根茎用于药用，栖息地退化，贸易和其他生物和人为干扰。一个有效的协议，利用间接和直接芽器官发生已优化为N。jatamansi。最好的愈伤组织实现了从切端的叶和叶柄外植体在MS培养基中补充1.5毫克/升α-萘乙酸和1.0毫克/升间-Topolin的培养15天内。在低温（13 +/-1 ° C）下培养外植体导致更好的愈伤组织生长、芽再生、超水力控制和再生芽中光合色素含量的改善。此外，茎尖和叶柄外植体的直接器官发生实现在MS培养基中含有1.0毫克/升的meta-Topolin。在补充有1.0 mg/l吲哚乙酸的相同培养基中获得最佳生根，其中诱导的平均根数为4.52根/芽。利用分子标记和流式细胞术对试管苗的遗传稳定性进行了评估，并与母株进行了比较。内含子剪接连接（ISJ）和起始密码子靶向多态性（SCoT）标记为基础的分析显示，在直接芽器官发生（DSO）衍生的植物，而愈伤组织介导的器官发生（CMO）衍生的植物的情况下，均匀的带型相比，母株表现出轻微的变化。N.发现jatamansi为2C = 1.40 +/- 0.01 pg，因此为684.6 Mbp（1C）。虽然器官发生的愈伤组织表现为混倍性，但在愈伤组织衍生的植物中，通过流式细胞仪检测没有主要的表型和遗传重排。显着较高的抗氧化活性，观察callusderived植物相比，母亲和DSO衍生的植物。植株部位、再生途径和不同的溶剂系统对试管苗中总酚、黄酮、生物碱、单宁含量的影响很大。
Nardostachys jatamansi (D. Don) DC., a small, perennial, rhizomatous herb of immense medicinal importance since ancient times, is restricted to specialized habitats of alpine Himalayas ranging from 3000 to 5200 m asl. The species has been recently listed as critically endangered under IUCN Red list of threatened species due to over exploitation of its rhizomes for medicinal uses, habitat degradation, trade and other biotic and anthropogenic interferences. An efficient protocol using both indirect and direct shoot organogenesis has been optimized for N. jatamansi. Best callusing was achieved from the cut ends of leaf and petiole explants within 15 days of culture in MS medium supplemented with 1.5 mg/l alpha-naphthalene acetic acid and 1.0 mg/l meta-Topolin. Culturing the explants at low temperature (13 +/- 1 degrees C) resulted in better callus growth, shoot regeneration, hyperhydricity control and improvement in photosynthetic pigment content in regenerated shoots. Also, direct organogenesis from shoot tip and petiole explants was achieved in MS medium containing 1.0 mg/l meta-Topolin. Optimum rooting was achieved in the same medium supplemented with 1.0 mg/l indole acetic acid wherein averages of 4.52 roots/shoot were induced. Genetic stability of in vitro-derived plantlets was assessed and compared to mother plant using molecular markers and flow cytometry. Intron Splice Junction (ISJ) and Start Codon Targeted polymorphism (SCoT) marker based profiling revealed uniform banding profile in case of direct shoot organogenesis (DSO)-derived plants while callus mediated organogenesis (CMO)-derived plants showed slight variations as compared to mother plant. The genome size of N. jatamansi was found to be 2C = 1.40 +/- 0.01 pg and therefore 684.6 Mbp (1C). Although organogenic calli showed mixoploidy but no major phenotypic and genetic rearrangements were detected by flow cytometry in callus-derived plants. Significantly higher antioxidant activity was observed in callusderived plants as compared to mother and DSO-derived plants. Plant parts, regeneration pathways and various solvent systems greatly affected the yields of total phenolics, flavonoids, alkaloids, tannins contents present in the in vitro raised plantlets.