Construction and Analysis of 2 Reciprocal Arabidopsis Introgression Line Populations

Abstract

Two new large reciprocal sets of introgression lines (ILs) were created between the Arabidopsis accessions Col-0 and C24. In both sets (78 ILs with Col-0 background and 62 ILs with C24 background), the donor segments cover almost the entire genome with an average substitution size of 18.3 cM. In addition to the basic sets of ILs, further subILs were developed for 2 genomic regions allowing better mapping resolution. SubILs carrying donor segments with candidate genes for flowering time and reduced fertility were used to demonstrate the usefulness of the reciprocal ILs for quantitative trait loci detection and fine mapping. For subIL development at high resolution around the reduced fertility locus, we used modified CelI-based assays in one-well format for both marker development and genotyping. This serves as a very flexible and cost-effective approach.

Allele mining and haplotype discovery in barley candidate genes for drought tolerance

“…Until now EcoTILLING has been used in plants to examine DNA variation in Arabidopsis (Comai et al 2004), in black cottonwood (Gilchrist et al 2006), in mung bean (Barkley et al 2008), in melon (Nieto et al 2007) and in barley in mlo and Mla resistance genes (Mejlhede et al 2006). In the present pilot study we have adopted and set up the EcoTILLING technology for detecting natural variation in barley CGs for drought tolerance according to the protocol described by Törjék et al (2008), which is based on the incorporation of fluorescently labeled dNTPs into the PCR products. An advantage of this kind of labeling is that fluorescently labeled primers are not required, making this approach more cost-effective.…”

“…PCR reactions and heteroduplex formation were performed using a PTC-200 thermal cycler (Bio-Rad Laboratories, Hercules, CA, USA) as follows: heat denaturation at 94°C for 2 min (10 min activation step for BioThermStar Hot Start Taq DNA Polymerase) followed by 40 polymerization cycles (denaturation: 94°C for 10 s, annealing: 55-60°C for 30 s, extension: 72°C for 1-3 min) and finally extension, denaturation and reannealing steps (72°C for 5 min, 99°C for 10 s, 70°C for 20 s, followed by 70 cycles of 70°C for 20 s decreasing 0.3°C per cycle). After PCR amplification, samples were digested either with CelI enzyme (provided by Georg Strompen, University of Potsdam, Germany) according to the protocol described by Törjék et al (2008) or with ENDO-1 (Serial Genetics, Evry, France) following the manufacturer's recommendations. After this step samples were precipitated with 15 ll isopropanol (centrifuged for 15 min at 4,000 rpm.…”

Hybrid Incompatibility in Arabidopsis Is Determined by a Multiple-Locus Genetic Network

“…Male-sterile C24 (Paul et al, 1992) and its seventh generation backcross to Col-0 (gifts from Rod Scott and Rinke Vinkenoog), whose transgenic male sterility is encoded between markers MSAT5.22 and MSAT5.10 on chromosome 5, were used as maternal parents to create F1s of NILs and two backcross (BC1) populations: BC1-Col-0 (80 individuals) and BC1-C24 (91 individuals). NILs were selected from 140 isogenic lines (Törjék et al, 2008). Plant growth was in a 16-h light cycle and 22°C.…”

Complex Quantitative Traits Cracked by the Mouse Inter-Subspecific Consomic Strains

“…By definition, each consomic strain has a single chromosome that was transferred from a donor strain into the genetic background of a host strain. Currently, consomic strains are constructed in many model organisms, such as Caenorhabditis elegans, Arabidopsis thaliana, and Zea mays, for the genetic dissection of various biological traits [8,54,59]. Mouse consomic strains are used for the dissection of genetic factors responsible for quantitative polygenic traits [17,30].…”