Haploid plants produced by centromeremediated genome elimination ravi, m. Researchers at the university of california davis have developed a novel method to produce haploid plants through seeds. One could also take steps to minimize outcrossing putting haploid plants in aerocons or even better using a bagging system. This was done by crossing the haploid inducer line with a plant that bypassed meiotic recombination and reduction. The passionate life of simon chan genome biology full text. Kapusi e, hensel g, coronado mj, broeders s, marthe c, otto i, kumlehn j. Haploid plants produced by centromeremediated genome. Composite interval mapping analysis of this population along with two established ril populations l er 2cvi0 and est1col0 revealed a number of shared and unique qtl. Genome elimination in arabidopsis thaliana can be caused by parental cenh3 differences mechanism of genome elimination caused by cenh3 alterations. The effect of cenh3 variation in the solanum genus on genome elimination is being studied in collaboration with the laboratory of roger chetelat. Simon chan publications university of california, davis. Validation of doubled haploid plants by enzymatic mismatch.
Altered centromere function has been implicated in genome elimination leading to formation of haploids plants. Centromere histone h3 and phospholipasemediated haploid. We identified plants with impaired centromere loading caused by a mutation in the centromeretargeting domain catd. Since the cellular origin of the plants produced is not. The effect of cenh3 variation in the solanum genus on genome elimination is being studied in collaboration with. The elimination of a selectable marker gene in the doubled haploid progeny of cotransformed barley plants. Marimuthu mp, jolivet s, ravi m, pereira l, davda jn, et al. Here we show that haploid arabidopsis thaliana plants can be easily generated through seeds by manipulating a single centromere protein, the centromerespecific histone cenh3 called cenpa.
Although it was initially discovered in arabidopsis, the approach has received a tremendous amount of attention in a short period of time, due to its dramatic potential for applications in plant breeding. Homozygous cenh3 mutant plants were confirmed by pcr genotyping using derived cleaved amplified polymorphic sequence dcaps oligos 5. Mar, 2012 the ability to construct doubled haploid populations via centromere mediated genome elimination synergizes with a highquality genome sequence and vast natural collections with corresponding genotypic information to strengthen a. Jun 04, 2019 centromere mediated genome elimination has also been exploited to convert a natural tetraploid arabidopsis into a diploid, reducing its ploidy to simplify breeding.
Doubled haploid technology is widely used in maize zea mays breeding. May 15, 2015 here, we describe extreme chromosomal restructuring during genome elimination, a process resulting from hybridization of arabidopsis plants expressing different centromere histones h3. Haploid plants produced by centromeremediated genome elimination. Editing tamtl gene induces haploid plants efficiently by. Sep 08, 2015 the generation of haploids is the most powerful means to accelerate the plantbreeding process. Cenh3 plays a universal role at eukaryote centromeres, so in principle this should be transferable to all plant species. Haploid plants, inheriting chromosomes from one parent only, have important advantages in genetic research but also crucially in plant breeding. Although the life cycle of most sexually reproducing plant species alternates between a diploid sporophytic phase and a highly reduced gametophytic haploid phase, it is possible to obtain haploid plants containing the same number of chromosomes in their somatic cells as do the normal gametes of the species dunwell, 2010. Chan, journalnature, year2010, volume464, pages615618. Point mutation impairs centromeric cenh3 loading and induces.
Haploid plants through seeds available technology for. In 2012, articles in pnas 4 and nature genetics 5 illustrated, respectively, the use of cenh3mediated genome elimination in the production of recombinant inbred lines and in molecular breeding. Aberrant alleles of cenh3 produce 330% haploid seed genome elimination only works after outcross some kind of disagreement of centromeres haploid plants produced by centromeremediated genome elimination nature, 2010. Centromeremediated genome elimination has also been exploited to convert a natural tetraploid arabidopsis into a diploid, reducing its ploidy to simplify breeding.
They can originate spontaneously in nature or as a result of various induction techniques. Haploids are plants sporophytes that contain a gametic chromosome number n. Wo2017081011a1 nontransgenic haploid inducer lines in. The advent of affordable, largescale dna sequencing methods, coupled with advanced computing power, is empowering a detailed analysis of the structure and function of chromosomes. In this phenomenon, one of the parental chromosome sets is completely lost during embryonic cell divisions, creating sterile haploid offspring. Catastrophic chromosomal restructuring during genome. Haploid plants produced by centromere mediated genome elimination nature, 464, 615618 2010. Centromeres are the chromosomal loci that attach to spindle. Haploid plants, inheriting chromosomes from one parent only, have important advantages in genetic research but also crucially in plant breeding, where they are used to create instant homozygous.
Genomeeliminationinducedbycenh3alterationsisnotspecific to the gfptailswap transgene. Cehn3 transgenes were used to complement the phenotype of cenh3 knockout and knock. According to the centromeresize model, cenh3mediated hi involves cross between a wildtype line and a haploid inducer line with smallerdefective centromeres. A doubled haploid ril population was first produced using centromeremediated genome elimination between accessions with long pro0 and intermediate col0 telomere lengths. Haploid plants produced by centromere mediated genome elimination. Frontiers using centromere mediated genome elimination to.
A wide range of methods has been developed to produce doubled haploid dh plants and recent advances promise efficient dh production in otherwise recalcitrant species. Panels il show nonreductional segregation 50 in haploid meiosis, forming haploid dyads l. Ascenh3isuniversalineukaryotes, our method may be extended to produce haploids in any plant species. Centromeremediated genome elimination is a fairly recent advancement in the production of haploid plant lines. The model of centromeremediated chromosome elimination. A competitive loading process between wt and mutant cenh3s occurs during the hybrid formation. Pdf haploid plants produced by centromeremediated genome. In plants, centromeremediated genome elimination is used to 361 produce maternal haploids kelliher et al. The cenh3gfptailswap haploid inducer line was described previously by ravi and chan, 2010. The ability to construct doubled haploid populations via centromeremediated genome elimination synergizes with a highquality genome sequence and vast natural collections with corresponding genotypic information to strengthen a. Further, doubled haploids are invaluable in basic genetic research such as gene discovery, genetic mapping, and genome sequencing, especially in perennial crops.
Uniparental genome elimination occurs in diverse taxa, including several distantly related plant species and even in fish 1, 2. Production of haploid plants that inherit chromosomes from only one parent can greatly accelerate plant breeding1, 2, 3. Doubled haploidy is a fundamental tool in plant breeding as it provides the fastest way to generate populations of meiotic recombinants in a genetically fixed state. The procedure of haploid inducer mediated genome editing imge in maize. A doubled haploid ril population was first produced using centromere mediated genome elimination between accessions with long pro0 and intermediate col0 telomere lengths. Mar 25, 2010 haploid plants, inheriting chromosomes from one parent only, have important advantages in genetic research but also crucially in plant breeding, where they are used to create instant homozygous. A crisprcas9 cassette could be introgressed or directly transformed into a haploid inducer hi line. Using centromere mediated genome elimination to elucidate the. Centromere mediated genome elimination has proven to be a powerful tool in arabidopsis genetics allowing generation of haploid plants, rapid production of recombinant inbred lines, and reverse breeding approaches ravi and chan, 2010. Shattered chromosomes are formed from the genome of the haploid inducer, consistent with genomic catastrophes affecting a single, laggard chromosome.
The mechanism by which chromosomes from one parent are selectively. Dec 12, 2018 doubled haploids offer a rapid method of producing homozygous lines for accelerated breeding of varieties and hybrids necessary to address the food demands of the next 23 decades. Haploids inherit chromosomes from one parent and when converted into perfect homozygotes by chromosome doubling become a powerful tool for plant breeding and genetics. Haploid plants produced by centromere mediated genome elimination ravi, m. Nov 07, 2017 panels il show nonreductional segregation 50 in haploid meiosis, forming haploid dyads l. Novel technologies in doubled haploid line development. Fertilization and uniparental chromosome elimination during. Generation of haploid plants and improved plant breeding. The analysis of 577 f1 plantlets obtained from crosses involving the hv. We elucidated whether point mutations in the centromere specific histone h3 variant cenh3 could be harnessed for the induction of haploids. Genomic instability, involving chromosome number and structure changes, has been documented in multiple systems. Maternal and paternal haploids can be generated through reciprocal crosses. Centromere mediated genome elimination is a fairly recent advancement in the production of haploid plant lines.
Haploid plants produced by centromeremediated genome elimination nature, 464, 615618 2010 chan, s. We identified plants with impaired centromere loading caused by a mutation in the centromere targeting domain catd. Read this article to learn about the production of haploid plants. Rapid creation of arabidopsis doubled haploid lines for. Haploid plants are characterized by possessing only a single set of chromosomes gametophytic number of chromosomes i. Uniparental genome elimination is typically achieved by crossing two sexually incompatible species, in some intraspeci. Two methods are generally used to produce haploids. Conventionally, haploid plants are generated by in vitro tissue culture of haploid plant gametophytes, pollen male, and embryo sac female. The generation of haploids is the most powerful means to accelerate the plantbreeding process. We elucidated whether point mutations in the centromerespecific histone h3 variant cenh3 could be harnessed for the induction of haploids. During genome elimination, the zygote the product of pollen sperm and egg fusion, inherits both parental chromosome sets but one of the two parental genomes is lost upon the following.
According to the centromere size model, cenh3 mediated hi involves cross between a wildtype line and a haploid inducer line with smallerdefective centromeres. Here, we describe extreme chromosomal restructuring during genome elimination, a process resulting from hybridization of arabidopsis plants expressing different centromere histones h3. The modified hi line was then used to pollinate an elite maize inbred line of any genotype. Haploids generated from a heterozygous individual and converted to diploid create instant homozygous lines, bypassing generations of inbreeding. Ravi and chan described a novel method of in vivo haploid induction through centromere.
Haploid production by centromere mediated genome elimination. The rapidly evolving centromerespecific histone has stringent functional requirements in arabidopsis thaliana genetics, 186, 461471 2010 ravi, m. Novel technologies in doubled haploid line development ren. Making haploid plants which inherit chromosomes from only one parent. Genetic architecture of natural variation of telomere length.
Doubled haploids offer a rapid method of producing homozygous lines for accelerated breeding of varieties and hybrids necessary to address the food demands of the next 23 decades. In a battle between parental chromosomes, a failure to reload. Apr 12, 2016 haploid plants produced by centromeremediated genome elimination. The invention further relates to a method for the production of haploid or doubled haploid plants and to the plants thus obtained. We have also exploited centromeremediated genome elimination to convert a natural tetraploid arabidopsis into a diploid, reducing its ploidy to simplify breeding. There are two approaches for the production of haploid plants.
In plants, centromere mediated genome elimination is used to 361 produce maternal haploids kelliher et al. Pdf haploids and doubled haploids in plant breeding. As cenh3 is universal in eukaryotes, our method may be extended to produce haploids in any plant species. Ravi m, chan sw 2010 haploid plants produced by centromeremediated genome elimination. Synthetic clonal reproduction through seeds science.
This method induces genome elimination from one parent in a cross with a precise mutation, rather than by culturing haploid cells or by crossing distantly related plants. Development of a haploidinducer mediated genome editing. Here, we describe a facile, nontissue culturebased in vivo method of haploid production through seeds in the model plant, arabidopsis thaliana. As cenh3 is universal in eukaryotes, our method can be extended to produce haploids in any plant species. Here we establish, through fine mapping, genome sequencing, genetic complementation, and gene editing, that haploid induction in maize zea mays is triggered by a frameshift mutation in. In plants, genome elimination resulting from certain interspecific crosses was described decades ago, but it is limited to occasional, natural occurrences. Apr 26, 2019 the model of centromere mediated chromosome elimination. In plants, haploid induction through genome elimination has recently been connected. Haploid plants produced by centromeremediated genome elimination nature, 464, 615618 2010. Spontaneous development of haploid plants has been known since 1922, when blakeslee first described this phenomenon in datura stramonium blakeslee et al. Centromere functional defects underlie genome elimination in barley. In plants, haploid induction through genome elimination has recently been connected mechanistically to. The present invention relates to a mutant plant of the cucurbitaceae family comprising a modified cenh3 gene, which mutant plant, when crossed to a wildtype plant having 2n chromosomes produces progeny, at least 0.
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