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首页> 美国卫生研究院文献>BioMed Research International >Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant
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Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant

机译：四叶紫杉的间接再生和SCoT，ISSR和RAPD标记对驯化植物遗传保真度的评估：濒危药用植物

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Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.
机译：四叶紫罗兰是重要的药用植物，因其具有重要的生物碱而闻名。在本研究中，我们报告了通过使用基于叶和茎的愈伤组织进行间接再生的标准化方案，通过体外克隆繁殖进行的保守性保存，并通过起始密码子靶向（SCoT），简单序列重复序列评估了适应植物的遗传保真度。 （ISSR）以及基于随机扩增多态性DNA（RAPD）标记的分析。最初在Murashige和Skoog（MS）培养基中补充了5.0μmg/ L，3.0μmg/ L的2,4-苯并呋喃，从叶，根和茎的外植体中诱导出最大量的易碎愈伤组织（按毫克计分别为378.7、323.8和412.8）。二氯苯氧基乙酸（2,4-D）和5.0）mg / L的萘乙酸（NAA）。在补充了TDZ（0.25 mg / L）+ BAP（2 mg / L）的MS培养基上，从叶片和老茧上获得具有最大芽芽数（25和20）的芽再生。再生的芽成功地生根，最大生根率为98.0，在IAA（1.0 mg / L）和IBA（1.0 mg / L）修正的全强度MS培养基上。使用2：1比例的无菌花园土壤和沙子硬化再生的小苗，然后在田间条件下进行驯化，使成活率达到86％。进行了基于SCoT，ISSR和RAPD引物的聚合酶链反应（PCR）分析，以检查与母本植物相比微繁殖植物中可能的遗传变异。在所使用的十种SCoT（S），ISSR（R）和RAPD（OPA）引物中，S2，R10和OPA3带有良好的DNA条带，具有良好的扩增能力。结果表明，再生植株与母株没有任何多态性。因此，体外再生的四叶红小豆幼苗被确认为真型。

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