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首页> 外文期刊>Journal of Biotechnology >Flow cytometric and morphological analyses of Pinus pinaster somatic embryogenesis
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Flow cytometric and morphological analyses of Pinus pinaster somatic embryogenesis

机译：松树体细胞胚发生的流式细胞仪和形态分析

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摘要

An approach combining morphological profiling and flow cytometric analysis was used to assess genetic stability during the several steps of somatic embryogenesis in Pinus pinaster. Embryogenic cell lines of P. pinaster were established from immature zygotic embryos excised from seeds obtained from open-pollinated trees. During the maturation stage, phenotype of somatic embryos was characterized as being either normal or abnormal. Based upon the prevalent morphological traits, different types of abnormal embryos underwent further classification and quantification. Nuclear DNA content of maritime pine using the zygotic embryos was estimated to be 57.04 pg/2C, using propidium iodide flow cytometry. According to the same methodology, no significant differences (P< or =0.01) in DNA ploidy were detected among the most frequently observed abnormal phenotypes, embryogenic cell lines, zygotic and normal somatic embryos, and somatic embryogenesis-derived plantlets. Although the differences in DNA ploidy level do not exclude the occurrence of a low level of aneuploidy, the results obtained point to the absence of major changes in ploidy level during the somatic embryogenesis process of this economically important species. Therefore, our primary goal of true-to-typeness was assured at this level.
机译：结合形态分析和流式细胞仪分析的一种方法被用来评估松树体细胞胚发生的几个步骤中的遗传稳定性。从未成熟的合子胚中建立了P. pinaster的胚性细胞系，该胚从从开放授粉的树中获得的种子中切除。在成熟阶段，体细胞胚的表型被表征为正常或异常。根据普遍的形态特征，对不同类型的异常胚胎进行了进一步的分类和定量。使用碘化丙啶流式细胞术，使用合子胚的海生松的核DNA含量估计为57.04 pg / 2C。根据相同的方法，在最常观察到的异常表型，胚胎发生细胞系，合子和正常体细胞胚以及体细胞胚发生衍生的小植株之间，未检测到DNA倍性的显着差异（P <或= 0.01）。尽管DNA倍性水平的差异并不排除非整倍性水平的降低，但是获得的结果表明，在该经济上重要的物种的体细胞胚发生过程中，倍性水平没有重大变化。因此，在这个级别上可以确保我们做到真实的首要目标。

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