Browsing by Person "Susic, Zoran"

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    Experimental and simulation studies on introgressing genomic segments from exotic into elite germplasm of rye (Secale cereale L.) by marker-assisted backcrossing
    (2005) Susic, Zoran; Geiger, Hartwig H.
    The introgression of exotic germplasm is a promising approach to increase the genetic diversity of elite rye breeding materials. Even though exotic germplasm may contain genomic segments that can improve oligo- and polygenically inherited traits, it has not been intensively utilized in modern rye breeding due to its agronomically inferior phenotypes and low performance level. Introgression of exotic germplasm requires techniques that would minimize negative side effects attributable to genetic interactions between recipient and donor. This appears achievable by the introgression library approach involving the systematic transfer of donor chromosome (DC) segments from an agriculturally unadapted source (donor) into an elite line (recipient, recurrent parent). A set of introgression lines (ILs) is thus developed, in which introgression is restricted to one or a few short DC segments. Ideally, the introgressed DC segments are evenly distributed over the whole recipient genome and the total genome of the exotic donor is comprised in the established set of ILs. The systematic development of an introgression library in rye has not been described yet. The main objectives of this study were to i) establish two rye introgression libraries by marker-assisted backcrossing, comprising of ILs each harbouring one to three DC segments and jointly covering most of the donor genome (DG), and ii) apply computer simulations to develop a highly effective and cost-efficient marker-assisted introgression strategy for the creation of introgression libraries in rye. A cross between a homozygous elite rye inbred line L2053-N (recurrent parent) and a heterozygous Iranian primitive rye population Altevogt 14160 (donor) was used as base material to generate the two libraries (F and G). Repeated backcrossing (BC) and subsequent selfing (S) until generation BC2S3 were chosen as the introgression method. The AFLP and SSR markers were employed to select individuals carrying desired DC segments, starting from generation BC1 to generation BC2S2. The chromosomal localization of DC segments, the number of DC segments per IL, and the proportion of recurrent parent genome were used as criteria to select parent individuals. This procedure resulted in the first two rye introgression libraries worldwide, comprising 40 BC2S3 ILs per library and covering 72% of the total DG in library F and 63% in library G (jointly approximately 80%). Most of the established ILs harboured one to three homozygous DC segments (on average 2.2 in both libraries), with a mean length of 18.3 cM in library F and 14.3 cM in library G. Computer simulations were conducted using the software PLABSIM version 2 to evaluate and optimize strategies for developing an introgression library in rye. Simulations were based on map-length estimates obtained from genotyping the BC1 generation of population F (7 chromosome pairs, genome size 665 cM). Six strategies differing in the number of BC and S generations were analysed, by setting the restrictions of sufficient DG coverage and RPG recovery. The medium-long BC3S1 strategy proved to be the most recommendable. It allows to achieve close to 100% DG coverage with moderate progeny sizes (19 individual per IL) in the individual generations and an acceptable total number of marker data points (52700), thus providing a good compromise between the cost and speed of an introgression procedure. Longer strategies are somewhat more cost-efficient but too time-demanding. The reverse is true for shorter strategies. An optimal allocation of resources is achieved by starting an introgression strategy with a small BC1 population (between 60 and 200 individuals) and stepwise increasing the progeny size per IL from about 15 to about 25-35 individuals in the succeeding generations. Targeting longer DC segments and using genetic maps with lower marker density allow a remarkable reduction in resources. This approach, however, possesses shortcomings when implementation in breeding is considered. The longer DC segments more likely carry i) unfavourable loci as well, ii) more than one gene controlling the trait in question, or iii) many additional loci affecting other traits. The major disadvantage of genetic maps with large marker distances is the unknown information about possible double cross-overs within marker intervals. All above-mentioned disadvantages may cause problems in the process of identification and isolation of genes controlling the trait of interest. Thus, a lower initial effort for the establishment of an introgression library will later on require additional efforts for using the ILs in breeding and genomics. Since the results of the simulation study became available after the marker-assisted establishment of the two rye introgression libraries had been finished, the dimensioning of the experimental study deviated from the optimum dimensioning determined in the simulation study: i) The BC2S2 introgression strategy was used in the empirical approach, whereas the BC3S1 strategy proved to be most recommendable in the simulation study. ii) The BC1 population sizes of libraries F and G (68 and 69, respectively) were far below the optimum value (200) determined in the simulation study for the chosen BC2S2 strategy. iii) The mean progeny sizes per IL from generation BC2 onwards varied between 7 and 21, whereas the optimum progeny size would have been two to three times higher. iv) The total number of analysed individuals (690 in library F, 684 in library G) was considerably lower than the optimum determined in the simulation study (3440). As a consequence, the coverage of the donor genome in the two libraries was incomplete and most ILs harboured more than a single DC segment. The potential application of the results of the simulation study would have increased the value of the developed ILs (higher DG coverage, lower number of DC segments per IL) considerably, despite limited resources. The effects of the introgressed DC segments on agronomically important qualitative and quantitative traits still need to be examined in multi-environmental field experiments. Introgression lines with beneficial DC segments may directly be used in practical hybrid rye breeding programs. Moreover, such ILs may be further backcrossed to create near isogenic lines (NILs) each carrying a single marker-characterized short DC segment. These NILs are an ideal starting point for high-resolution mapping and for the isolation and functional characterisation of candidate genes. The two rye introgression libraries and the results of the simulation study mark important milestones for the targeted exploitation of exotic rye germplasm and provide a promising opportunity to proceed towards functional genomics in rye.