Le Signor Christine, Savois Vincent, Aubert Grégoire, Verdier Jérôme, Nicolas Marie, Pagny Gaelle, Moussy Françoise, Sanchez Myriam, Baker Dave, Clarke Jonathan, Thompson Richard
Unité Mixte de Recherche en Génétique et Ecophysiologie des Légumineuses à Graines, Institut National de la Recherche Agronomique, Dijon, France.
Plant Biotechnol J. 2009 Jun;7(5):430-41. doi: 10.1111/j.1467-7652.2009.00410.x.
Medicago truncatula has been widely adopted as a model plant for crop legume species of the Vicieae. Despite the availability of transformation and regeneration protocols, there are currently limited tools available in this species for the systematic investigation of gene function. Within the framework of the European Grain Legumes Integrated Project (http://www.eugrainlegumes.org), chemical mutagenesis was applied to M. truncatula to create two mutant populations that were used to establish a TILLING (targeting induced local lesions in genomes) platform and a phenotypic database, allowing both reverse and forward genetics screens. Both populations had the same M2 line number, but differed in their M1 population size: population 1 was derived from a small M1 population (one-tenth the size of the M2 generation), whereas population 2 was generated by single seed descent and therefore has M1 and M2 generations of equal size. Fifty-six targets were screened, 10 on both populations, and 546 point mutations were identified. Population 2 had a mutation frequency of 1/485 kb, twice that of population 1. The strategy used to generate population 2 is more efficient than that used to generate population 1, with regard to mutagenesis density and mutation recovery. However, the design of population 1 allowed us to estimate the genetically effective cell number to be three in M. truncatula. Phenotyping data to help forward screenings are publicly available, as well as a web tool for ordering seeds at http://www.inra.fr/legumbase.
蒺藜苜蓿已被广泛用作蚕豆族作物豆科植物的模式植物。尽管已有转化和再生方案，但目前该物种中可用于系统研究基因功能的工具仍然有限。在欧洲谷物豆类综合项目（http://www.eugrainlegumes.org）的框架内，对蒺藜苜蓿进行了化学诱变，以创建两个突变群体，用于建立一个TILLING（靶向诱导基因组局部损伤）平台和一个表型数据库，从而实现反向和正向遗传学筛选。两个群体的M2系编号相同，但M1群体大小不同：群体1来自一个较小的M1群体（M2代大小的十分之一），而群体2是通过单粒传产生的，因此M1和M2代大小相等。共筛选了56个靶点，两个群体各10个，共鉴定出546个点突变。群体2的突变频率为1/485 kb，是群体1的两倍。就诱变密度和突变回收率而言，用于产生群体2的策略比用于产生群体1的策略更有效。然而，群体1的设计使我们能够估计蒺藜苜蓿中遗传有效细胞数为3。有助于正向筛选的表型数据可公开获取，同时还有一个用于订购种子的网络工具，网址为http://www.inra.fr/legumbase。
