Department of Plant and Environmental Sciences, Göteborg University, S-40530, Göteborg, Sweden.
BMC Plant Biol. 2010 May 12;10:86. doi: 10.1186/1471-2229-10-86.
Oat, Avena sativa is the sixth most important cereal in the world. Presently oat is mostly used as feed for animals. However, oat also has special properties that make it beneficial for human consumption and has seen a growing importance as a food crop in recent decades. Increased demand for novel oat products has also put pressure on oat breeders to produce new oat varieties with specific properties such as increased or improved beta-glucan-, antioxidant- and omega-3 fatty acid levels, as well as modified starch and protein content. To facilitate this development we have produced a TILLING (Targeting Induced Local Lesions IN Genomes) population of the spring oat cultivar SW Belinda.
Here a population of 2600 mutagenised M2 lines, producing 2550 M3 seed lots were obtained. The M2 population was initially evaluated by visual inspection and a number of different phenotypes were seen ranging from dwarfs to giants, early flowering to late flowering, leaf morphology and chlorosis. Phloroglucinol/HCl staining of M3 seeds, obtained from 1824 different M2 lines, revealed a number of potential lignin mutants. These were later confirmed by quantitative analysis. Genomic DNA was prepared from the M2 population and the mutation frequency was determined. The estimated mutation frequency was one mutation per 20 kb by RAPD-PCR fingerprinting, one mutation per 38 kb by MALDI-TOF analysis and one mutation per 22.4 kb by DNA sequencing. Thus, the overall mutation frequency in the population is estimated to be one mutation per 20-40 kb, depending on if the method used addressed the whole genome or specific genes. During the investigation, 6 different mutations in the phenylalanine ammonia-lyase (AsPAL1) gene and 10 different mutations in the cellulose synthase-like (AsCslF6) beta-glucan biosynthesis gene were identified.
The oat TILLING population produced in this work carries, on average, hundreds of mutations in every individual gene in the genome. It will therefore be an important resource in the development of oat with specific characters. The population (M5) will be available for academic research via Nordgen http://www.nordgen.org as soon as enough seeds are obtained.[Genbank accession number for the cloned AsPAL1 is GQ373155 and GQ379900 for AsCslF6].
燕麦，Avena sativa 是世界上第六大重要的谷物。目前，燕麦主要用作动物饲料。然而，燕麦也具有特殊的性质，使其有益于人类食用，并在最近几十年作为一种粮食作物越来越受到重视。对新型燕麦产品的需求增加也给燕麦育种者带来了压力，要求他们生产具有特殊性质的新型燕麦品种，如增加或改善β-葡聚糖、抗氧化剂和 ω-3 脂肪酸水平，以及改性淀粉和蛋白质含量。为了促进这一发展，我们培育了春燕麦品种 SW Belinda 的 TILLING（靶向诱导基因组局部突变）群体。
这里获得了一个由 2600 个诱变 M2 系组成的群体，产生了 2550 个 M3 种子批。M2 群体最初通过目视检查进行评估，观察到各种不同的表型，范围从矮化到巨人，从早花到晚花，叶片形态和黄化。从 1824 个不同的 M2 系获得的 M3 种子的 phloroglucinol/HCl 染色显示了一些潜在的木质素突变体。这些后来通过定量分析得到证实。从 M2 群体中提取基因组 DNA，并确定突变频率。通过 RAPD-PCR 指纹分析，每 20kb 有一个突变，通过 MALDI-TOF 分析，每 38kb 有一个突变，通过 DNA 测序，每 22.4kb 有一个突变，估计突变频率为每 20-40kb 一个突变。因此，根据所使用的方法是否涉及整个基因组或特定基因，该群体的总体突变频率估计为每 20-40kb 一个突变。在研究过程中，鉴定了苯丙氨酸氨裂解酶（AsPAL1）基因中的 6 种不同突变和纤维素合酶样（AsCslF6）β-葡聚糖生物合成基因中的 10 种不同突变。
本工作中产生的燕麦 TILLING 群体在基因组的每个个体基因中平均携带数百个突变。因此，它将成为开发具有特定特性的燕麦的重要资源。一旦获得足够的种子，该群体（M5）将通过 Nordgen http://www.nordgen.org 提供给学术研究使用[克隆的 AsPAL1 的 Genbank 登录号为 GQ373155 和 GQ379900 用于 AsCslF6]。
