Doubled Haploid Technology: Generation of Doubled Haploid Maize Lines Using Haploid Inducers.

Vencke K Grüning, Thomas Lübberstedt, Ursula K Frei
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Abstract

Doubled haploid (DH) technology allows for the development of completely homozygous lines from heterozygous plants in only two generations. This approach has been widely adopted in maize breeding programs, as it expedites the generation of inbred lines compared to traditional methods. The DH approach is based on the use of maize genotypes that have the ability to induce haploid seeds when used as the pollen parent. The most common method for producing maize haploid plants for the generation of DH lines is in vivo maternal haploid induction. The process involves pollination with a haploid inducer maize line to generate haploid seeds. Then, haploids are screened for and identified (typically via the expression of a particular marker gene), germinated, treated with an exogenous doubling agent to induce genome duplication, and transplanted to the field. Following successful self-pollination, seeds harvested from the ear represent fully homozygous lines. The seed set at this stage, however, is often low, necessitating one or two additional rounds of self-pollination to increase the number of fully homozygous inbred lines. Here, we describe a protocol for the generation of maize DH lines using maternal haploid-inducing maize lines. We outline the steps for setting up the donor material, performing induction crosses, selecting haploids based on two different marker alleles, treating seedlings with colchicine to double the genome, transplanting the treated seedlings to the field, and self-pollinating the treated plants.

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双单倍体技术:使用单倍体诱导剂生成双倍单倍体玉米品系。
双单倍体(DH)技术只需两代就能从杂合植株培育出完全同源的品系。与传统方法相比,这种方法能加快近交系的产生,因此已被玉米育种计划广泛采用。DH 方法的基础是使用具有诱导单倍体种子能力的玉米基因型作为花粉亲本。生产玉米单倍体植株以培育近交系的最常用方法是体内母本单倍体诱导。这一过程包括用单倍体诱导玉米品系授粉,产生单倍体种子。然后,对单倍体进行筛选和鉴定(通常是通过特定标记基因的表达)、发芽、用外源加倍剂处理以诱导基因组复制,并移植到田间。自花授粉成功后,从果穗上收获的种子代表完全同源的品系。但这一阶段的结实率往往很低,因此需要再进行一到两轮自花授粉,以增加完全同源的近交系数量。在此,我们介绍一种利用母本单倍体诱导玉米品系培育玉米 DH 品系的方法。我们概述了建立供体材料、进行诱导杂交、根据两种不同的标记等位基因选择单倍体、用秋水仙素处理幼苗使基因组加倍、将处理过的幼苗移植到田间以及对处理过的植株进行自花授粉的步骤。
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Cold Spring Harbor protocols
Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.00
自引率
0.00%
发文量
163
期刊介绍: Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.
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