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通过玻璃化法进行超低温保存的桃棕(Bactris gasipaes, Kunth)体细胞胚的存活及超微结构特征

Survival and ultrastructural features of peach palm (Bactris gasipaes, Kunth) somatic embryos submitted to cryopreservation through vitrification.

作者信息

Heringer Angelo Schuabb, Steinmacher Douglas André, Schmidt Éder Carlos, Bouzon Zenilda Laurita, Guerra Miguel Pedro

机构信息

Graduate Program in Plant Genetic Resources, Plant Developmental Physiology and Genetics Laboratory, Federal University of Santa Catarina, Rod. Admar Gonzaga, Km 3, Florianópolis, SC, 88034-001, Brazil.

出版信息

Protoplasma. 2013 Oct;250(5):1185-93. doi: 10.1007/s00709-013-0500-4. Epub 2013 May 1.

Abstract

Bactris gasipaes (Arecaceae), also known as peach palm, was domesticated by Amazonian Indians and is cultivated for its fruit and heart-of-palm, a vegetable grown in the tree's inner core. Currently, the conservation of this species relies on in situ conditions and field gene banks. Complementary conservation strategies, such as those based on in vitro techniques, are indicated in such cases. To establish an appropriate cryopreservation protocol, this study aimed to evaluate the ultrastructural features of B. gasipaes embryogenic cultures submitted to vitrification and subsequent cryogenic temperatures. Accordingly, somatic embryo clusters were submitted to Plant Vitrification Solution 3 (PVS3). In general, cells submitted to PVS3 had viable cell characteristics associated with apparently many mitochondria, prominent nucleus, and preserved cell walls. Cells not incubated in PVS3 did not survive after the cryogenic process in liquid nitrogen. The best incubation time for the vitrification technique was 240 min, resulting in a survival rate of 37 %. In these cases, several features were indicative of quite active cell metabolism, including intact nuclei and preserved cell walls, an apparently many of mitochondria and lipid bodies, and the presence of many starch granules and condensed chromatin. Moreover, ultrastructure analysis revealed that overall cellular structures had been preserved after cryogenic treatment, thus validating the use of vitrification in conjunction with cryopreservation of peach palm elite genotypes, as well as wild genotypes, which carry a rich pool of genes that must be conserved.

摘要

桃棕(棕榈科),也被称为桃椰,由亚马逊印第安人驯化,因其果实和棕榈心(一种生长在树内芯的蔬菜)而被种植。目前,该物种的保护依赖于原地条件和田间基因库。在这种情况下,需要采取补充性的保护策略,比如基于体外技术的策略。为了建立合适的超低温保存方案,本研究旨在评估经玻璃化处理及随后的低温处理的桃棕胚性培养物的超微结构特征。因此,将体细胞胚簇置于植物玻璃化溶液3(PVS3)中。一般来说,经PVS3处理的细胞具有存活细胞的特征,有明显较多的线粒体、突出的细胞核以及完整的细胞壁。未在PVS3中孵育的细胞在液氮低温处理后未能存活。玻璃化技术的最佳孵育时间为240分钟,存活率为37%。在这些情况下,一些特征表明细胞代谢相当活跃,包括完整的细胞核和完整的细胞壁、明显较多的线粒体和脂质体,以及许多淀粉粒和凝聚染色质的存在。此外,超微结构分析表明,低温处理后细胞的整体结构得以保存,从而验证了玻璃化结合超低温保存桃棕优良基因型以及野生基因型的方法的有效性,这些野生基因型携带了必须保存的丰富基因库。

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