A rich TILLING resource for studying gene function in Brassica rapa.
A rich TILLING resource for studying gene function in Brassica rapa.
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DOI：
10.1186/1471-2229-10-62
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发表时间：
2010-04-09
影响因子：
5.3
通讯作者：
Østergaard L
中科院分区：
文献类型：
--
作者：
Stephenson P;Baker D;Girin T;Perez A;Amoah S;King GJ;Østergaard L
十字花科植物包括模式植物拟南芥以及一些重要的农业物种，如油料作物(特别是油菜、芥菜和油菜)和蔬菜(如油菜、油菜和油菜)。白菜和甘蓝)。十字花科植物与拟南芥仅相隔1000-2000万年，它们之间的亲缘关系十分密切，有关拟南芥生长发育的知识有望转化为十字花科作物，用于作物改良。此外，植物发育的某些方面在油菜和拟南芥之间有很大的不同，因此有必要直接在作物物种中进行研究。然而，双二倍体甘蓝型油菜和芥菜型油菜中单个基因的突变可能不会产生预期的表型，因为这些物种的基因组中每个单拷贝的拟南芥基因最多可以包含六个同源基因。因此，为了阐明和可能开发冗余基因在油菜作物改良中的作用，在甘蓝型油菜等二倍体甘蓝型油菜中研究冗余基因的作用可能更有效。此外，白菜基因组正在进行的测序使该物种成为甘蓝研究和遗传资源开发的一个极具吸引力的模式。用甲基磺酸乙酯(EMS)处理甘蓝型油菜二倍体A基因组种的种子，产生TILL(靶向诱导的基因组局部损伤)群体，用于反向遗传学研究。我们使用了白菜型油菜，R-o-18，它具有与油菜作物相似的发育个体发育。因此，这一资源有望很好地用于研究与油菜产量和品质相关的性状。从总共9,216株M2植株中提取DNA，并汇集在一起，形成耕作平台的基础。对6个基因的分析显示，突变水平很高，密度约为每60kb一个。这一分析还表明，仅在三分之一的群体(3072个M2植株)中筛选1 kb的扩增子将提供平均68个突变和97%的概率获得导致截短蛋白的终止密码子突变。我们进一步计算了每种植物平均含有~10,000个突变，由于植物的数量很大，预计基因组中大约一半的GC碱基对的突变存在于这个群体中。我们在二倍体甘蓝型油菜中开发了第一个EMS分蘖资源。该种群的突变密度为每60kb~1个，这使其成为已公布分耕种群的突变最密集的二倍体生物。该资源可通过RevGenUK反向遗传学平台http://revgenuk.jic.ac.uk.公开获得
The Brassicaceae family includes the model plant Arabidopsis thaliana as well as a number of agronomically important species such as oilseed crops (in particular Brassica napus, B. juncea and B. rapa) and vegetables (eg. B. rapa and B. oleracea). Separated by only 10-20 million years, Brassica species and Arabidopsis thaliana are closely related, and it is expected that knowledge obtained relating to Arabidopsis growth and development can be translated into Brassicas for crop improvement. Moreover, certain aspects of plant development are sufficiently different between Brassica and Arabidopsis to warrant studies to be carried out directly in the crop species. However, mutating individual genes in the amphidiploid Brassicas such as B. napus and B. juncea may, on the other hand, not give rise to expected phenotypes as the genomes of these species can contain up to six orthologues per single-copy Arabidopsis gene. In order to elucidate and possibly exploit the function of redundant genes for oilseed rape crop improvement, it may therefore be more efficient to study the effects in one of the diploid Brassica species such as B. rapa. Moreover, the ongoing sequencing of the B. rapa genome makes this species a highly attractive model for Brassica research and genetic resource development. Seeds from the diploid Brassica A genome species, B. rapa were treated with ethyl methane sulfonate (EMS) to produce a TILLING (Targeting Induced Local Lesions In Genomes) population for reverse genetics studies. We used the B. rapa genotype, R-o-18, which has a similar developmental ontogeny to an oilseed rape crop. Hence this resource is expected to be well suited for studying traits with relevance to yield and quality of oilseed rape. DNA was isolated from a total of 9,216 M2 plants and pooled to form the basis of the TILLING platform. Analysis of six genes revealed a high level of mutations with a density of about one per 60 kb. This analysis also demonstrated that screening a 1 kb amplicon in just one third of the population (3072 M2 plants) will provide an average of 68 mutations and a 97% probability of obtaining a stop-codon mutation resulting in a truncated protein. We furthermore calculated that each plant contains on average ~10,000 mutations and due to the large number of plants, it is predicted that mutations in approximately half of the GC base pairs in the genome exist within this population. We have developed the first EMS TILLING resource in the diploid Brassica species, B. rapa. The mutation density in this population is ~1 per 60 kb, which makes it the most densely mutated diploid organism for which a TILLING population has been published. This resource is publicly available through the RevGenUK reverse genetics platform http://revgenuk.jic.ac.uk.
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影响因子：
56.9
作者：
Alonso, JM;Stepanova, AN;Ecker, JR
通讯作者：
Ecker, JR
影响因子：
14.9
作者：
Oleykowski, CA;Mullins, CRB;Yeung, AT
通讯作者：
Yeung, AT
DOI：
10.1007/1-4020-3735-x_20
发表时间：
2005-01-01
期刊：
LOTUS JAPONICUS HANDBOOK
影响因子：
--
作者：
Perry, Jillian;Welham, Tracey;Wang, Trevor
通讯作者：
Wang, Trevor
影响因子：
3.3
作者：
Parkin, IAP;Gulden, SM;Lydiate, DJ
通讯作者：
Lydiate, DJ
影响因子：
3.8
作者：
Rusholme, Rachel L.;Higgins, Erin E.;Lydiate, Derek J.
通讯作者：
Lydiate, Derek J.