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在雌雄异株的宽叶蝇子草中,X染色体对于体细胞发育是必需的:细胞遗传学和分子证据以及单倍体基因组测序。

The X chromosome is necessary for somatic development in the dioecious Silene latifolia: cytogenetic and molecular evidence and sequencing of a haploid genome.

作者信息

Soukupova Magda, Nevrtalova Eva, Cížková Jana, Vogel Ivan, Cegan Radim, Hobza Roman, Vyskot Boris

机构信息

Department of Plant Developmental Genetics, Institute of Biophysics ASCR, Brno, Czech Republic.

出版信息

Cytogenet Genome Res. 2014;143(1-3):96-103. doi: 10.1159/000363431. Epub 2014 Jun 27.

Abstract

Silene latifolia (or white campion) possesses a well-established sex determination system with a dominant Y chromosome in males (the mammalian type). The heteromorphic sex chromosomes X and Y in S. latifolia largely stopped recombination; thus, we can expect a gradual genetic degeneration of the Y chromosome. It is well proven that neither diploid nor polyploid S. latifolia sporophytes can survive without at least one X, so the only life stage possessing the Y as the sole sex chromosome is the male gametophyte (pollen tube), while the female gametophyte seems to be X-dependent. Previous studies on anther-derived plants of this species showed that the obtained plants (largely haploid or dihaploid) were phenotypically and cytologically female. In this paper, we provide molecular evidence for the inviability of plants lacking the X chromosome. Using sex-specific PCR primers, we show that all plantlets and plants derived from anther cultures are female. In studying anther-derived diploid females by sequencing of X-linked markers, we demonstrate that these plants are really homozygous dihaploids. A haploid regenerant plant was sequenced (8× genome coverage) using Illumina technology. Genome data are disposable in the EMBL database as a standard for full genome and X chromosome assembly in this model species. Homozygous dihaploids were back-crossed with males to yield a progeny useful for the study of the evolution of the Y chromosome.

摘要

宽叶蝇子草(或白剪秋萝)拥有一套完善的性别决定系统,雄性具有一条显性Y染色体(哺乳动物类型)。宽叶蝇子草中异形性染色体X和Y基本停止了重组;因此,我们可以预期Y染色体会逐渐发生基因退化。充分证实,无论是二倍体还是多倍体的宽叶蝇子草孢子体,若没有至少一条X染色体都无法存活,所以唯一以Y作为唯一性染色体的生命阶段是雄配子体(花粉管),而雌配子体似乎依赖于X染色体。此前对该物种花药衍生植株的研究表明,所获得的植株(大多为单倍体或双单倍体)在表型和细胞学上均为雌性。在本文中,我们提供了缺乏X染色体的植株无法存活的分子证据。使用性别特异性PCR引物,我们表明所有来自花药培养的幼苗和植株均为雌性。通过对X连锁标记进行测序来研究花药衍生的二倍体雌性植株,我们证明这些植株确实是纯合双单倍体。使用Illumina技术对一株单倍体再生植株进行了测序(基因组覆盖度为8倍)。基因组数据已作为该模式物种全基因组和X染色体组装的标准存入EMBL数据库。将纯合双单倍体与雄性植株回交,以产生可用于研究Y染色体进化的后代。

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