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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Rose protoplast isolation and culture and heterokaryon selection by immobilization in extra thin alginate film
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Rose protoplast isolation and culture and heterokaryon selection by immobilization in extra thin alginate film

机译：固定在超薄藻酸盐膜中的玫瑰原生质体的分离和培养以及异核体的选择

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摘要

Somatic hybridization has been identified as one method for the genetic improvement of roses. The success of somatic hybridization programmes relies to a great extent upon efficient protoplast isolation and culture and selection of heterokaryons. This paper reports the isolation of rose cell suspension protoplasts by direct sucrose flotation and demonstrates their culture using extra thin alginate film. A comparative assessment of the efficiency of conventional culture techniques versus those with extra thin alginate film or thin alginate layer is also presented. A very high plating efficiency (80%) was obtained using thin alginate layer or extra thin alginate film techniques with improved media formulations. Protoplasts of Rosa damascena and R. bourboniana were fused by using polyethylene glycol as fusogen and later immobilized in the thin layer of alginate. The fused protoplasts were tracked on the basis of differential fluorescent staining, and the hybridity of heterokaryons following their development to callus was confirmed by molecular characterization. This novel selection strategy has general applicability and is faster and simpler to perform during somatic hybridization experiments.
机译：体细胞杂交已被确定为玫瑰遗传改良的一种方法。体细胞杂交程序的成功在很大程度上取决于有效的原生质体分离以及异核体的培养和选择。本文报道了通过直接蔗糖浮选分离玫瑰细胞悬液原生质体的方法，并证明了使用超薄藻酸盐薄膜进行培养的情况。还提供了对传统培养技术与超薄藻酸盐膜或藻酸盐薄层的效率的比较评估。使用薄藻酸盐层或超薄藻酸盐膜技术以及改进的介质配方，可以获得非常高的电镀效率（80％）。罗莎大马士革和波旁亚种的原生质体通过使用聚乙二醇作为融合剂进行融合，然后固定在藻酸盐薄层中。融合的原生质体在荧光差异染色的基础上进行了追踪，并通过分子表征证实了杂核体发展为愈伤组织后的杂交性。这种新颖的选择策略具有普遍的适用性，并且在体细胞杂交实验中执行起来更快，更简单。

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