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Applicability of the droplet vitrification method to olive somatic embryos

机译：液滴玻璃化法对橄榄体细胞胚的适用性

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摘要

Somatic embryos of olive have been successfully cryopreserved using the droplet-vitrification method on aluminium foil strips (1) after dehydration in PVS2 for 30 min. Although acceptable recovery rates have been obtained (2) and cultures morphology apparently did not change, the influence of this cryopreservation protocol on the somatic embryogenesis process is unknown. The aim of this investigation was to assess the influence of the droplet-vitrification method on the different phases of somatic embryogenesis and to evaluate its applicability to somatic embryos of different genotypes.The results obtained revealed that the genotype plays a key role in the maintenance and maturation phases and determines the regeneration potential of the embryogenic lines. Cryopreservation only affected some specific aspects, such as the proliferation capacity in maintenance medium, germination of newly developed embryos and length of the shoots obtained. Nevertheless, a statistically significant interaction between genotype and cryopreservation was appreciated in different parameters related to cultures proliferation. The influence of cryopreservation on cultures morphology during the maturation phase and embryo germination was also genotype-dependent.Thus, the regeneration potential was not significantly affected by cryopreservation and the quality of the plantlets obtained was similar in both types of cultures, although slightly longer shoots were obtained in plants derived from cryopreserved explants.In relation to the applicability of the droplet-vitrification method to olive somatic embryos of different origin, response to cryopreservation significantly varied depending of the genotype, with recovery rates ranging from 0 to 60%. The appearance of cultures established from frozen somatic embryos was similar to that of the corresponding controls.In conclusion, our investigations show that droplet-vitrification is a reliable procedure for preserving the viability, embryogenic competence and regeneration capacity of olive somatic embryos. Nevertheless, additional optimization of this method is required in order to improve the recovery rates obtained in some of the genotypes tested.(1) Panis B, Piette B & Swennen R (2005) Plant Sci. 168, 45-55. (2) Sánchez-Romero C & Bradaï F (2013) in Abstract Book X Reunión SECIVTV, pp 56.
机译：在PVS2中脱水30分钟后，已使用液滴玻璃化方法在铝箔条（1）上成功冷冻了橄榄的体细胞胚。尽管已经获得了可接受的恢复率（2），并且培养物的形态显然没有改变，但是这种低温保存方案对体细胞胚发生过程的影响尚不清楚。这项研究的目的是评估液滴玻璃化方法对体细胞胚发生的不同阶段的影响，并评估其对不同基因型的体细胞胚胎的适用性。结果表明，基因型在维持和维持胚乳中起着关键作用。成熟阶段，并确定胚系的再生潜力。冷冻保存仅影响某些特定方面，例如在维持培养基中的增殖能力，新发育的胚的发芽和获得的芽的长度。然而，在与培养物增殖有关的不同参数中，基因型和冷冻保存之间的统计学上显着的相互作用被赞赏。冷冻保存对成熟期和胚胎萌发过程中培养物形态的影响也是基因型依赖性的。因此，冷冻保存对再生潜能没有显着影响，尽管两种苗的芽长略长，但两种培养物的幼苗质量相似从冷冻外植体衍生的植物中获得。关于液滴玻璃化方法对不同来源的橄榄体细胞胚的适用性，对冷冻保存的响应随基因型而显着变化，回收率在0％至60％之间。从冷冻的体细胞胚建立的培养物的外观与相应的对照相似。总之，我们的研究表明，液滴玻璃化是保存橄榄体细胞胚的活力，胚胎发生能力和再生能力的可靠方法。尽管如此，仍需要对该方法进行其他优化，以提高在某些测试的基因型中获得的回收率。（1）Panis B，Piette B和Swennen R（2005）Plant Sci。 168，45-55。 （2）Sánchez-RomeroC和BradaïF（2013）在《 SEC XTV XReunión》摘要第56页中。

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