Reference standards for determination of DNA content of plant nuclei
Reference standards for determination of DNA content of plant nuclei
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DOI：
10.2307/2656569
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发表时间：
1999-05-01
影响因子：
3
通讯作者：
Price, HJ
中科院分区：
文献类型：
--
作者：
Johnston, JS;Bennett, MD;Price, HJ
采用流式细胞术对14种植物DNA含量测定标准品进行比较。采用鸡和植物内标，碘化丙啶（PI）和4'-6-二氨基-2-苯基吲哚（DAPI）作为荧光染料。给出了14个潜在参考标准的平均值和标准误差，并与Feulgen密度测定法的平均值进行了比较。建议将五种植物作为未来植物DNA含量测定的初步国际标准：高粱；先锋8695 (2C = 1.74 pg), Pisum sativum cv。Minerva Maple (2C = 9.56 pg), Hordeum vulgare cv。苏丹（2C = 11.12 pg），蚕豆（2C = 26.66 pg）和葱属植物(cv。艾尔莎·克雷格（2C = 33.55 pg）。建议选择2C和JC核DNA含量峰与靶种2C和4C峰相似但不重叠的参比标准品。我们推荐PI作为流式细胞术测定植物DNA含量的首选荧光染料。只有当估计的DNA值通过使用第二种染色证实时，DAPI才应使用，该染色对基因组中富含AT或cc的序列没有偏倚。
Flow cytometry was used to compare 14 potential reference standards for plant DNA content determination. Both chicken and plant internal standards were used, as were propidium iodide (PI) and 4'-6-diamidino-2-phenylindole (DAPI) as fluorochromes. Means and standard errors of the means are presented for the 14 potential reference standards, and the means are compared to those obtained by Feulgen densitometry. Five species are recommended as an initial set of international standards for future plant DNA content determinations: Sorghum bicolor cv. Pioneer 8695 (2C = 1.74 pg), Pisum sativum cv. Minerva Maple (2C = 9.56 pg), Hordeum vulgare cv. Sultan (2C = 11.12 pg), Vicia faba (2C = 26.66 pg), and Allium cepa cv. Ailsa Craig (2C = 33.55 pg). It is recommended that the reference standard of choice be one with 2C and JC nuclear DNA content peaks similar to, but not overlapping, the 2C and 4C peaks of the target species. We recommend PI as the fluorochrome of choice for flow cytometric determination of plant DNA content. DAPI should be used only if the estimated DNA Value is corroborated by using a second stain that has no bias for AT- or CC-rich sequences within genomes.