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研究水稻卵细胞中线粒体形态和 DNA 含量。

Studies of mitochondrial morphology and DNA amount in the rice egg cell.

机构信息

Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.

出版信息

Curr Genet. 2010 Feb;56(1):33-41. doi: 10.1007/s00294-009-0277-3. Epub 2009 Dec 5.

Abstract

In plant vegetative cells, mitochondria are usually small and grain-shaped. In contrast, unusually shaped giant mitochondria (large cup-shaped or long stretched-rod-shaped) appear in the egg cells of geranium, maize, Ipomoea nil, and bracken. In this study, to characterize egg cell mitochondria in rice, we used nonenzymatic manual dissection to isolate unfertilized egg cells of rice and observed the egg cell mitochondria and mitochondrial DNA (mtDNA) simultaneously. These observations showed that the mitochondria in the rice egg cell are small and grain-shaped, unlike the mitochondria in geranium, maize, I. nil, and bracken. Double staining of mitochondria by MitoTracker and mtDNA by SYBR Green I showed that mitochondria in the rice egg cell have a large amount of mtDNA compared with the rice root protoplast. We also used real-time PCR analysis to quantify the mtDNA amount in the rice egg cell. We quantified the copy numbers of four mitochondrial genes per single rice egg cell and rice leaf protoplast. Real-time PCR analysis revealed that the egg cell has more than ten times more copy numbers of all of four genes encoded in the mitochondrial genome compared with the leaf protoplast.

摘要

在植物的营养细胞中,线粒体通常呈小颗粒状。相比之下,天竺葵、玉米、蕹菜和蕨类植物的卵细胞中会出现形状异常的巨型线粒体(大杯状或长拉伸棒状)。在这项研究中,为了研究水稻卵细胞中的线粒体,我们使用非酶促手动分离方法分离水稻未受精卵细胞,并同时观察卵细胞线粒体和线粒体 DNA(mtDNA)。这些观察结果表明,水稻卵细胞中的线粒体呈小颗粒状,与天竺葵、玉米、蕹菜和蕨类植物中的线粒体不同。用 MitoTracker 对线粒体进行双重染色和用 SYBR Green I 对 mtDNA 进行双重染色表明,与水稻根原生质体相比,水稻卵细胞中的线粒体含有大量的 mtDNA。我们还使用实时 PCR 分析定量水稻卵细胞中的 mtDNA 含量。我们对每个水稻卵细胞和水稻叶原生质体中的四个线粒体基因进行了实时 PCR 分析,定量了每个基因的拷贝数。实时 PCR 分析显示,与叶原生质体相比,四个基因的所有基因在卵细胞中的拷贝数都多了十倍以上。

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