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Evaluating a DOE screen to reduce hyperhydricity in the threatened plant, Cycladenia humilis var. jonesii
In Vitro Cellular & Developmental Biology - Plant ( IF 2.6 ) Pub Date : 2020-01-02 , DOI: 10.1007/s11627-019-10038-y
Valerie C. Pence , Linda R. Finke , Randall P. Niedz

In vitro methods for propagation and preservation are critical when seed banking is not an option for the ex situ conservation of an endangered species. Such methods rely on the ability to maintain normal growth for the ultimate goal of producing plants. Previous studies revealed that the threatened Cycladenia humilis var. jonesii can exhibit extreme hyperhydricity when cultured in vitro, but that this phenotype can be largely normalized by venting of the cultures. A design of experiments (DOE) approach was used to investigate factors contributing to hyperhydricity in this species. Using a 6-factor fractional factorial design, the effects of nitrogen, CaCl2, BAP, gelling agent, and the presence of venting on the phenotype were tested. Venting was the strongest driver decreasing hyperhydricity, but gelling agent and other factors had significant effects, depending on the response measured. A revised medium was then designed and compared with the original maintenance medium through 2–3 subcultures. Venting was again the strongest driver decreasing hyperhydricity, but the revised medium also significantly affected some responses, even in the absence of venting. The revised medium was not optimal, as shoots became progressively smaller with each transfer, but despite their size, hyperhydricity decreased. These results demonstrate the usefulness of DOE for simultaneous testing of multiple factors, but also the need for follow-up optimization experiments. DOE can be particularly useful in working with rare species. By making efficient use of limited resources, it can facilitate the development of in vitro propagation protocols for such species and help ensure their conservation.



中文翻译:

评估DOE筛选以减少濒临灭绝的植物Cycladenia humilis var。琼斯

当种子库不是濒危物种的非原生境保存的选择时,繁殖和保存的体外方法至关重要。此类方法依赖于维持正常生长的能力以达到生产植物的最终目的。先前的研究表明,濒临灭绝的Cycladenia humilis var。当在体外培养时jonesii可能表现出极高的水合作,但是这种表型可以通过排出培养物而在很大程度上归一化。实验设计(DOE)方法用于研究促成该物种高水度的因素。使用六因子分数阶乘设计,氮,CaCl 2的影响,BAP,胶凝剂和排气在表型上的存在进行了测试。排气是降低高水压的最强驱动力,但是胶凝剂和其他因素有显着影响,具体取决于测得的反应。然后设计了经过修改的培养基,并通过2-3次亚培养与原始的维持培养基进行了比较。放气再次成为降低高水压的最强驱动力,但是即使没有放气,修订后的培养基也显着影响了某些反应。改良的培养基不是最佳的,因为每次转移都使芽逐渐变小,但是尽管其大小,高水合度却降低了。这些结果证明了DOE对于同时测试多个因素的有用性,但也需要后续优化实验。DOE在处理稀有物种时特别有用。通过有效利用有限的资源,可以促进发展这些物种的体外繁殖方案,并有助于确保其保存。

更新日期:2020-01-02
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