The Recent Development in Technologies for Attaining Doubled Haploid Plants In Vivo

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Abstract

Haploid plants with a doubled set of chromosomes (doubled haploid (DH)) significantly speed up the selection process by the fixation of genetic traits in each locus in the homozygous state within one generation. Doubled haploids are mainly attained by the formation of plants from the cultured gametophytic (haploid) tissues and cells in vitro, or by targeted reduction in the parent chromosome during intra- or interspecific hybridization. Since then, DH has become one of the most powerful tools to support various basic research studies, as well as applied research. This review is focused on the recent development of the production of doubled haploids in vivo and their fundamental bases. The various mechanisms and approaches responsible for the formation of haploids in vivo are discussed, particularly the induction of parthenogenesis by BBM-like proteins, the long constructed Salmon system of wheat, the usage of patatin-like phospholipases MTL/PLA1/NLD, the IG1 system, uniparental genome elimination during interspecific hybridization, and the perspective technology of centromeric histone 3 (CENH3) modification.

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DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins.

Je Woo, Jungeun Kim, Soon Kwon … (2015)

Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

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Haploid plants produced by centromere-mediated genome elimination.

Maruthachalam Ravi, Simon W L Chan (2010)

Production of haploid plants that inherit chromosomes from only one parent can greatly accelerate plant breeding. Haploids generated from a heterozygous individual and converted to diploid create instant homozygous lines, bypassing generations of inbreeding. Two methods are generally used to produce haploids. First, cultured gametophyte cells may be regenerated into haploid plants, but many species and genotypes are recalcitrant to this process. Second, haploids can be induced from rare interspecific crosses, in which one parental genome is eliminated after fertilization. The molecular basis for genome elimination is not understood, but one theory posits that centromeres from the two parent species interact unequally with the mitotic spindle, causing selective chromosome loss. Here we show that haploid Arabidopsis thaliana plants can be easily generated through seeds by manipulating a single centromere protein, the centromere-specific histone CENH3 (called CENP-A in human). When cenh3 null mutants expressing altered CENH3 proteins are crossed to wild type, chromosomes from the mutant are eliminated, producing haploid progeny. Haploids are spontaneously converted into fertile diploids through meiotic non-reduction, allowing their genotype to be perpetuated. Maternal and paternal haploids can be generated through reciprocal crosses. We have also exploited centromere-mediated genome elimination to convert a natural tetraploid Arabidopsis into a diploid, reducing its ploidy to simplify breeding. As CENH3 is universal in eukaryotes, our method may be extended to produce haploids in any plant species.

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Ectopic expression of BABY BOOM triggers a conversion from vegetative to embryonic growth.

Thérèse Ouellet, Lemin Zhang, Manuel Van Domselaar … (2002)

The molecular mechanisms underlying the initiation and maintenance of the embryonic pathway in plants are largely unknown. To obtain more insight into these processes, we used subtractive hybridization to identify genes that are upregulated during the in vitro induction of embryo development from immature pollen grains of Brassica napus (microspore embryogenesis). One of the genes identified, BABY BOOM (BBM), shows similarity to the AP2/ERF family of transcription factors and is expressed preferentially in developing embryos and seeds. Ectopic expression of BBM in Arabidopsis and Brassica led to the spontaneous formation of somatic embryos and cotyledon-like structures on seedlings. Ectopic BBM expression induced additional pleiotropic phenotypes, including neoplastic growth, hormone-free regeneration of explants, and alterations in leaf and flower morphology. The expression pattern of BBM in developing seeds combined with the BBM overexpression phenotype suggests a role for this gene in promoting cell proliferation and morphogenesis during embryogenesis.