NRC Publications Archive Archives des publications du CNRC Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production
NRC Publications Archive Archives des publications du CNRC Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production
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游离小孢子培养物为研究小孢子诱导和胚胎发生提供了一个极好的系统，为不断增加的分子研究阵列提供了一个平台，并且可以产生用于加速植物育种计划的双单倍体（DH）植物。此外，分离的小孢子培养物具有优于花药培养物的几个优点，其中花药壁的存在可导致二倍体、体细胞愈伤组织和植物的发育。尽管不同实验室的游离小孢子培养方案不同，但培养供体植物、收获花器官、分离小孢子、培养和诱导小孢子、再生胚和染色体加倍的基本步骤保持相同。在过去的几年里，关于游离小孢子培养的研究报告中有很大一部分集中在谷类和芸苔属物种上。对于其中一些物种，游离小孢子培养方案已经建立，并在世界各地的实验室中常规使用，用于开发新品种，以及基因组学、基因表达和遗传图谱等领域的基础研究。尽管这些物种被认为对小孢子培养高度敏感，但效率仍在提高。然而，对于许多物种，孤立的小孢子培养在产生DH植物方面还不够有效，从而对于育种计划来说是成本有效的。最近，单倍体研究重新兴起，一些曾经被认为顽固的物种也出现了反应。未来的研究计划，旨在阐明参与小孢子诱导和胚胎发生的途径将是有益的，因为将新的方法来提高小孢子培养DH生产的效率。对于许多物种，花药培养已被证明比游离小孢子培养更有效，需要更多的研究来阐明花药壁对胚胎发生的贡献。开发分子标记用于确定再生植物的配子起源，而不管其倍性如何，也将是有益的。在这篇综述中，我们的目的是提供一个概述的基本游离小孢子培养协议，重点是最近的进展，在几种作物。nigra（L.）、、和几种B基因型。napus（Ferrie and Keller 2007）. B。欧洲油菜小孢子方案已经成功地扩展到另一种十字花科物种，Leskovšek等人（2008）报道了从芝麻菜或芝麻菜（Eruca sativa Mill.）生产DH小植株，一年生多叶蔬菜。
An isolated microspore culture provides an excellent system for the study of microspore induction and embryogenesis, provides a platform for an ever-increasing array of molecular studies, and can produce doubled haploid (DH) plants, which are used to accelerate plant-breeding programs. Moreover, isolated microspore cultures have several advantages over anther culture, wherein presence of the anther walls can lead to the development of diploid, somatic calli and plants. Although protocols for isolated microspore culture vary from laboratory to laboratory, the basic steps of growing donor plants, harvesting floral organs, isolating microspores, culturing and inducing microspores, regenerating embryos, and doubling the chromosomes, remain the same. Over the past few years, a large proportion of the research reports on isolated microspore culture have focused on cereal and Brassica species. For some of these species, isolated microspore culture protocols are well established and routinely used in laboratories around the world for developing new varieties, as well as for basic research in areas such as genomics, gene expression, and genetic mapping. Although these species are considered highly responsive to microspore culture, improvements in efficiency are still being made. However, with many species, isolated microspore culture is simply not yet efficient enough at producing DH plants to be cost-effective for breeding programs. There has been a recent resurgence of haploidy research with response being reported in some species once considered recalcitrant. Future research programs aimed at elucidating pathways involved in microspore induction and embryogenesis will be of benefit, as will novel approaches to improve the efficiency of microspore culture for DH production. With many species, anther culture has proven to be more effective than isolated microspore culture, necessitating more research to clarify the contribution of the anther wall to embryogenesis. The development of molecular markers for use in determining the gametic origin of regenerated plants, irrespective of their ploidy, would also be beneficial. In this review, we aim to provide an overview of the basic isolated microspore culture protocol with an emphasis on recent progress in several crop species. nigra (L.) , , and several genotypes of B. napus (Ferrie and Keller 2007). The B. napus microspore protocol has been successfully extended to another cruciferous species with the report by Leskovšek et al. (2008) of DH plantlet production from rocket or arugula ( Eruca sativa Mill.), a leafy, annual vegetable.