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Reverse breeding: a novel breeding approach based on engineered meiosis - PubMed

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Reverse breeding: a novel breeding approach based on engineered meiosis

Rob Dirks et al. Plant Biotechnol J. 2009 Dec.

Free PMC article

Abstract

Reverse breeding (RB) is a novel plant breeding technique designed to directly produce parental lines for any heterozygous plant, one of the most sought after goals in plant breeding. RB generates perfectly complementing homozygous parental lines through engineered meiosis. The method is based on reducing genetic recombination in the selected heterozygote by eliminating meiotic crossing over. Male or female spores obtained from such plants contain combinations of non-recombinant parental chromosomes which can be cultured in vitro to generate homozygous doubled haploid plants (DHs). From these DHs, complementary parents can be selected and used to reconstitute the heterozygote in perpetuity. Since the fixation of unknown heterozygous genotypes is impossible in traditional plant breeding, RB could fundamentally change future plant breeding. In this review, we discuss various other applications of RB, including breeding per chromosome.

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Figures

Figure 1
Figure 1

Reverse breeding can be used to fix unknown heterozygotes. Crossing two homozygous parents (grey and black bars) creates a heterozygous F1. When selfed, the F1 produces a segregating F2 population. A starting hybrid of unknown genetic constitution is selected for its desireable characteristics, and subjected to the two steps of reverse breeding (grey box). By knocking down meiotic crossing over, whole parental chromosomes are transmitted through spores, without rearrangement. Note, in this example the four chromosomes in the hybrid can generate 16 different combinations in the gametes—only five are shown for convenience. The achiasmatic gametes are then used produce doubled haploid (DH) lines using in vitro culture techniques. From this population, complementary parents can be chosen that when crossed perfectly reconstitute the starting hybrid. The DH lines then serve as a permanent library that can be used to predictably generate a wide variety of defined hybrids.

Figure 2
Figure 2

Reverse breeding can be used as advanced breeding tool. As starting hybrid for a reverse breeding experiment, a fully heterozygous F1 is chosen, resulting from a cross between two homozygous parents. Application of reverse breeding (grey box) leads to a population of doubled haploids. Note that among those DHs, there are chromosome substitution lines of one of the starting parents into the backgroud of the other. Lower left: a chromosome substitution line for a grey chromosome in the black parent can be backcrossed with the fully black parent to create a hybrid that is heterozygous for just one chromosome. Such hybrids serve as starting point for breeding per chromosome (explained in text). Lower right: a chromosome substitution line for a black chromosome in the grey parent can be backcrossed with the fully black parent to create a hybrid that is homozygous for just one chromosome. Such hybrids are starting points for studying background interactions (explained in text).

Figure 3
Figure 3

The presence of a single crossover in a chromosome pair does not affect the utility of reverse breeding. The figure depicts four cells at different stages of meiosis. At metaphase I, a single crossover is present in one chromosome pair (a bivalent pair) whereas other homologues remain as univalents. The homologues joined by a chiasma segregate to opposite poles and—in this example—the univalents segregate randomly to opposite poles, giving rise, in this case, to a balanced dyad (at telophase I). Meiosis then proceeds through metaphase II, separating sister chromatids, and at telophase II four gametes are formed. Half of these gametes contain a recombinant chromosome (upper two), whereas the other half contain non-recombinant chromosomes (lower two) and are useful for reverse breeding.

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