Genetic Control of Embryo Lethality in Crosses between Common Wheat and Rye

The phenotypic manifestation and genetic control of embryo lethality observed in crosses between common wheat and rye were studied. It was found that crosses between common wheat and inbred self-fertile rye lines L2 and 535 gave rise to ungerminating grains, in which the development and differentiation of the hybrid embryo are arrested. Study of the degree of embryo development in the hybrid grains obtained by crossing common wheat varieties with inbred rye lines L2 and 535 showed that genotypes of the parents affected the ratio between undifferentiated embryos of various sizes. Analysis of this trait was performed by test crosses according to a novel pedigree program with the use of interlinear hybrids and a set of fourth-generation hybrid recombinant inbred lines. Rye line L2 was shown to bear the Eml (Embryo lethality) gene, which terminates the development of the hybrid embryo in amphihaploids. The suggestion of complementary interaction between wheat and rye genes during formation of a “new” character in wheat-rye F₁ hybrids is discussed. A method of detecting an allele not complementary to the rye Eml allele in wheat is proposed. The proposed test program allows appropriate study of the system of wheat and rye genes involved in complementary interaction in the genotype of a distant hybrid.__________Translated from Genetika, Vol. 41, No. 8, 2005, pp. 1075–1083.Original Russian Text Copyright © 2005 by Tikhenko, Tsvetkova, Voylokov.     

Gene mutations in rye causing embryo lethality in hybrids with wheat: allelism and chromosomal localisation

In crosses between hexaploid wheat and inbred lines of rye, a small number of rye genotypes produce seeds carrying undifferentiated, non-viable embryos. Hybrids between such lines and those not showing this phenotype were used as pollen donors in crosses with bread wheat in order to determine the genetic basis of disturbed embryo development. A single gene, designated Eml-R1b, is causing this character. Molecular markers associated with F₂ genotypes derived from a contrasting rye inbred progeny were used for a linkage study. Recombinant inbred lines of an F₅ population served as testers. Eml-R1b maps to chromosome arm 6RL, along with two co-segregating microsatellite loci, Xgwm1103 and Xgwm732. Complementary interactions of deleterious genes in wheat and rye are discussed.     

Features of crossability, haploidy and polyembryony in hybrid combinations between common barley Hordeum vulgare L. (2n = 14) and wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya

The role of individual chromosomes of rye in the manifestation of crossability and seedling development in hybrid combinations between common barley Hordeum vulgare L., cultivar Nepolegayushchii (2n = 14) and five wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya (2n = 40 wheat + 2 rye chromosomes). Crossability, which was measured by two parameters--frequency of set grains and frequency of grains with embryos--was shown to be significantly affected by each of the five rye chromosomes examined: 1R, 2R, 3R, 5R, and 6R; the development of barley haploids was affected by rye chromosomes 1 R, 3R, and 5R. We were the first to demonstrate that polyembryony could be induced by mutual effects of barley cytoplasm and rye chromosome 1R. Possible mechanisms controlling the development of haploids and twins in hybrid combinations H. vulgare x T. aestivum/S. cereale are discussed. The conclusion is drawn that hybrid combinations between common barley and wheat-rye substitution lines can serve as new models for studying incompatibility mechanisms in distant crosses and genetic control of parthenogenesis.     

Hybrid dwarfness in crosses between wheat (Triticum aestivum L.) and rye (Secale cereale L.): a new look at an old phenomenon

The existence of hybrid dwarfs from intraspecific crosses in wheat (Triticum aestivum) was described 100 years ago, and the genetics underlying hybrid dwarfness are well understood. In this study, we report a dwarf phenotype in interspecific hybrids between wheat and rye (Secale cereale). We identified two rye lines that produce hybrid dwarfs with wheat and have none of the hitherto known hybrid dwarfing genes. Genetic analyses revealed that both rye lines carry a single allelic gene responsible for the dwarf phenotype. This gene was designated Hdw‐R1 (Hybrid dwarf‐R1). Application of gibberellic acid (GA₃) to both intraspecific (wheat–wheat) and interspecific (wheat–rye) hybrids showed that hybrid dwarfness cannot be overcome by treatment with this phytohormone. Histological analysis of shoot apices showed that wheat–rye hybrids with the dwarf phenotype at 21 and 45 days after germination failed to develop further. Shoot apices of dwarf plants did not elongate, did not form new primordia and had a dome‐shaped appearance in the seed. The possible relationship between hybrid dwarfness and the genes responsible for the transition from vegetative to generative growth stage is discussed.     

Embryo lethality in wheat-rye hybrids: dosage effect and deletion bin mapping of the responsible wheat locus

The speciation allele at Eml-A1 of hexaploid wheat, which causes embryo lethality in wheat-rye hybrids, was investigated using cytologically modified genetic stocks. It was demonstrated that an extra dose of this allele had no effect on embryo development in these hybrids. There was no positive effect on embryo development and, therefore, no overcoming of the postzygotic barrier. An abortion of the hybrid embryos at an earlier stage of development was also not observed. Physical mapping was performed using chromosome 6A deletion lines. This study revealed the location of Eml-A1 on the most distal part of the long arm of chromosome 6A. To identify possible candidate genes responsible for embryo lethality, in silico sequence homology analysis was performed. Two candidate genes for Eml-A1 that are involved in shoot apical meristem maintenance were identified on chromosome 6AL. However, functional validation assays need to be designed and performed.     

Contributions of heterosis and epistasis to hybrid fitness

Early-generation hybrid fitness is difficult to interpret because heterosis can obscure the effects of hybrid breakdown. We used controlled reciprocal crosses and common garden experiments to distinguish between effects of heterosis and nuclear and cytonuclear epistasis among morphotypes and advanced-generation hybrid derivative populations in the Piriqueta caroliniana (Turneraceae) plant complex. Seed germination, growth, and sexual reproduction of first-generation hybrids, inbred parental lines, and outbred parental lines were compared under field conditions. Average vegetative performance was greater for hybrids than for inbred lines, and first-season growth was similar for hybrids and outbred parental lines. Hybrid survival surpassed that of inbred lines and was equal to or greater than outbred lines' survival, and more F(1) than parental plants reproduced. Reductions in hybrid fitness due to Dobzhansky-Muller incompatibilities (epistasis among divergent genetic elements) were expressed as differences in vegetative growth, survival, and reproduction between plants from reciprocal crosses for both F(1) and backcross hybrid generations. Comparing performance of hybrids against parental genotypes from intra- and interpopulation crosses allowed a more robust prediction of F(1) hybrids' success and more accurate interpretations of the genetic architecture of F(1) hybrid vigor.     

Mechanisms and inheritance of first division restitution in hybrids of wheat, rye, and Aegilops squarrosa.

First division restitution (FDR) in intergeneric Triticeae hybrids provides an important meiotic mechanism for the production of amphidiploids without the use of colchicine and similar chemicals. The genetic controls of FDR were investigated by examining microsporogenesis and fertility in F1 hybrids of two- and three-way crosses of durum wheat (Triticum turgidum L. var. durum) cultivars Langdon (LDN) and Golden Ball (GB), 'Gazelle' rye (Secale cereale L.), and one accession (RL5286) of Aegilops squarrosa L. The results from two-way crosses indicated that the first meiotic division varied, depending on the hybrid. GB crossed with Ae. squarrosa developed tripolar spindles and prevented congregation of chromosomes at the equatorial plate. The hybrid of GB with rye had a delayed first division. But, the hybrids of LDN with both Ae. squarrosa and rye had a high frequency of FDR. Analysis from the three-way crosses indicated that inheritance in rye crosses differed from those with Ae. squarrosa. FDR segregated in a 1:1 ratio in the rye cross, suggesting that the FDR is controlled by a single gene from LDN. However, FDR fit a 1:3 ratio in the three-way crosses with Ae. squarrosa. Cytological data suggested that tripolar spindles are a major factor preventing FDR in Ae. squarrosa crosses. Some progenies from the three-way cross with rye had a high frequency of monads that resulted from second division failure of FDR cells.     

Features of Crossability, Haploidy and Polyembryony in Hybrid Combinations between Cultivated Barley Hordeum vulgare L. (2n = 14) and Wheat-Rye Substitution Lines Triticum aestivum L., Cultivar Saratovskaya 29/Secale cereale L., Cultivar Onokhoiskaya

The role of individual chromosomes of rye in the manifestation of crossability and seedling development in hybrid combinations between cultivated barley Hordeum vulgare L., cultivar Nepolegayushchii (2n = 14) and five wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya (2n = 40 wheat + 2 rye chromosomes). Crossability, which was measured by two parameters—frequency of set grains and frequency of grains with embryos—was shown to be significantly affected by each of the five rye chromosomes examined: 1R, 2R, 3R, 5R, and 6R; the development of barley haploids was affected by rye chromosomes 1R, 3R, and 5R. We were the first to demonstrate that polyembryony could be induced by mutual effects of barley cytoplasm and rye chromosome 1R. Possible mechanisms controlling the development of haploids and twins in hybrid combinations H. vulgare × T. aestivum/S. cereale are discussed. The conclusion is drawn that hybrid combinations between cultivated barley and wheat-rye substitution lines can serve as new models for studying incompatibility mechanisms in distant crosses and genetic control of parthenogenesis.__________Translated from Genetika, Vol. 41, No. 6, 2005, pp. 784–792.Original Russian Text Copyright © 2005 by Pershina, Belova, Devyatkina, Rakovtseva, Kravtsova, Shchapova.     

Analysis of the effects of parental genotypes of rye lines on the development of quantitative traits in primary octaploid triticale. Plant height

When breeding the primary spring octoploid triticale derived from crosses of various inbred rye lines to wheat Chinese Spring, the effects of the rye genotype and growth conditions on the plant height and proportion of the first, second, and final (pedicle) internodes to the entire stem length were studied. Two triticale groups were examined: homozygotes for the dominant (Ddw1) and recessive (ddw1) alleles of the gene responsible for short stem in rye. In the short stem triticale lines carrying the Ddw1 alleles, the plants were 20 cm shorter on average than those in the ddw1-carrying lines, and the distribution of the two triticale groups overlapped significantly. In both groups, the lines significantly differing in plant height could be differentiated, because of allelic diversity of the additional genes controlling this trait along with the Ddw gene. In most triticale lines, especially in the Ddw1-carrying ones, the plant height was much reduced under unfavorable growth conditions. At the same time, a short-stem line was isolated, which is characterized by ecological plasticity, like the maternal wheat cultivar. In the triticale studied, the stem structure depended on the short-stem rye genotype. The two-year study showed that in the triticale carrying the dominant allele of this gene, the first internode is significantly extended, whereas the upper (pedicle) internode is reduced, which increases plant lodging resistance. The differences revealed between the rye lines as well as their effect on the quantitative triticale traits are discussed in view of a variant of the hybridological analysis, which had been previously proposed for identification and mapping of the correspondent rye genes.     

Oligonucleotides replacing the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 for FISH analysis

Hybrids derived from wheat (Triticum aestivum L.) × rye (Secale cereale L.) have been widely studied because of their important roles in wheat cultivar improvement. Repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 are usually used as probes in fluorescence in situ hybridization (FISH) analysis of wheat, rye, and hybrids derived from wheat × rye. Usually, some of these repetitive sequences for FISH analysis were needed to be amplified from a bacterial plasmid, extracted from bacterial cells, and labeled by nick translation. Therefore, the conventional procedure of probe preparation using these repetitive sequences is time-consuming and labor-intensive. In this study, some appropriate oligonucleotide probes have been developed which can replace the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 in FISH analysis of wheat, rye, and hybrids derived from wheat × rye. These oligonucleotides can be synthesized easily and cheaply. Therefore, FISH analysis of wheat and hybrids derived from wheat × rye using these oligonucleotide probes becomes easier and more economical.     

Effect of genetic diversity in Hordeum vulgare L. and Secale cereale L. on their crossability and the development and viability of hybrid embryos and plants

A study was made of the effect of genetic diversity in cultivated barley and rye on seed setting, embryonic development, and viability of barley x rye hybrids grown in vitro and planted. The variation in seed-setting frequency was shown to be determined mainly by vegetation conditions and random factors, while the variation in proportion of seeds with embryos among the total seed set was determined by the genetic diversity of barley and rye. Selection of barley genotypes was shown to increase the yield of viable barley x rye hybrids. Based on the phenotypic analysis of barely x rye hybrids obtained in various combinations, low viability and hybrid lethality were attributed to the hybrid necrosis genes expressed from early ontogeny. These genes are assumed to be widespread among barley and rye varieties.     

The effect of wheat cytoplasm on the synthesis of wheat x rye hybrids

Summary Four nuclear genotypes of Triticum aestivum L., each substituted into the cytoplasm of T. timococcum Kostoff, Aegilops ventricosa Tausch and T. timopheevi Zhuk., were crossed with four homozygous Secale cereale L. inbred lines to produce 48 alloplasmic octoploid primary triticales. Crossability, embryo differentiation, amphihaploid plantlet recovery in vitro, and response to colchicine treatment were investigated. It was found that in general the effects of the wheat cytoplasm on the characteristics studied were of equal importance as those of the wheat and the rye nuclear genotypes. Cytoplasm can be regarded as an additional source of variation in the synthesis of primary triticales. Cross-compatibility in wheat x rye hybrids appeared to be determined by specific parental genotypes and their nucleo-cytoplasmic interactions rather than by any general compatibility of particular nuclear genotypes and/or cytoplasms.     

Absence of heterosis in hybrid corn. Description of the effect

Three unrelated homozygous maize lines, TS11, P22, and ST156, that produced hybrids in which heterosis was either absent or insignificant were identified. These hybrids were phenotypically similar to self-pollinated homozygous lines. Reciprocal crosses showed that the absence of heterosis is controlled by nuclear genes and is not associated with the cytoplasm of inbred lines. Analysis of F2 plants demonstrated that lines TS11, P22, and ST156 contained allelic genes determining the absence of heterosis in hybrid plants. Crosses of lines TS11, P22, and ST156 with a common selection line 092 generated hybrids with normal heterosis. It was concluded that heterozygosity or homozygosity of particular genes in lines TS11, P22, and ST156 play a pivotal role in the manifestation or the absence of hybrid vigor in hybrids.     

rRNA gene activity and control of expression mediated by methylation and imprinting during embryo development in wheat x rye hybrids

Ribosomal RNA genes originating from one parent are often suppressed in interspecific hybrids. We show that treatments during germination with the cytosine analogue 5-azacytidine stably reactivate the expression of the suppressed rRNA genes of rye origin in the wheat x rye amphiploid, triticale, by preventing methylation of sites in the rye rDNA. When 5-azacytidine is applied to embryos of triticale and wheat x rye F₁ hybrids nine, or more, days after fertilization, rye rRNA gene expression is stably reactivated in the resulting seedling. Earlier treatments have no effect on rye rRNA gene expression, indicating that undermethylation of DNA early in embryo development is reversible. After 9 days, the methylation status of rRNA genes in maintained throughout development. Since the change in expression follows a methylation change at particular restriction-enzyme sites, the data establish a clear correlation between gene activity and methylation in plants.     

Genetic interactions between wheat and rye genomes in triticale

Summary Six primary triticale lines were produced from two advanced breeding lines of Triticum durum and three inbred genotypes of Secale cereale. The wheat and rye parents and the triticale derivatives were crossed in all possible combinations within each species group. Chiasma and univalent frequency of parents and hybrids were determined. The primary triticale lines had more univalents and less chiasmata per pollen mother cell than the corresponding wheat and rye parents together. The parental wheat F₁ exhibited negative heterosis for chiasma frequency whereas all rye hybrids had much higher chiasma frequencies than their inbred parents. Triticale F₁s generally showed lower chiasma frequencies and more univalents than their parents, but the degree of pairing failure was dependent upon which of the parental species within the triticale, wheat or rye, was in the heterozygous state. F₁s with heterozygous wheat genome only showed the least reduction in chiasma number (presumably caused by gene actions within the wheat genome), while F₁s with heterozygous rye genome showed high reduction in chiasma frequency and an increase in pairing failure (induced by negative interactions between the heterozygous rye and the wheat genome in triticale). A high correlation was found between the frequency of undisturbed pollen mother cells and the frequency of aneuploids in the subsequent generation. A higher number of aneuploids occurred in those populations which were heterozygous for the rye genome.     

Agronomic Trait and Protein Component of F2 Hybrid Originated from Intergeneric Somatic Hybridization Between Triticum aestivumand Agropyron elongatum

Protoplasts derived from common wheat ( Triticum aestivum L． cv. Jinan 177) were fused with UV-treated protoplasts of Agropyron elongatum (Host) Nevski by PEG method, and fertile asymmetric somatic hybrid plants resembling wheat morphology were obtained. The F2 hybrid plants could be divided into 3 types according to their morphology. Type Ⅰ hybrids had high and loosely standing stalks with big spikes and grains. TypeⅡ hybrids were dwarf and compact in shape with high tillering ability and smaller spikes. Type Ⅲ hybrids were similar to typeⅠas a whole but had more compact and erect spikes. All the F2 hybrid lines were superior to wheat in seed protein content, although some difference existed between themselves. Protein analysis of immature embryos and flag leaves from hybrids by two-dimensional electrophoresis showed that they possessed characteristic proteins of both parents and some new proteins. There existed also some different kinds of proteins in different lines.     

The genetic and molecular characterization of pollen-derived plant lines from octoploid triticale x wheat hybrids

Six doubled-haploid (DH) lines, derived by anther culture from octoploid triticale x wheat hybrids, were characterized using cytological, biochemical and molecular techniques. Lines varied in their wheat and rye genome composition, and were either wheat-rye chromosome multiple addition lines or had spontaneous substitutions and/or wheat-rye translocations. Most of the lines contained a pair of 4R chromosomes, whereas 1R or 7R were present in others. The results are similar to those previously obtained with hexaploid triticale x wheat crosses and indicate that it is possible to produce alien (wheat/rye) addition, substitution, and translocation lines directly from the anther culture of intergeneric hybrids.     

Effect of rye Secale cereale L. chromosomes 1R and 3R on polyembryony expression in hybrid combinations between (Hordeum vulgare L.)-Triticum aestivum L. alloplasmic recombinant lines and wheat T. aestivum L.-rye S. cereale L. substitution lines

The effect of rye chromosomes on polyembryony was studied for reciprocal hybrid combinations between (Hordeum vulgare L.)-Triticum aestivum L. alloplasmic recombinant lines and five wheat T. aestivum L. (cultivar Saratovskaya 29)-rye Secale cereale L. (cultivar Onokhoiskaya) substitution lines: IR(1D), 2R(2D), 3R(3B), 5R(5A), and 6R(6A), and for direct hybrid combinations between the [H. marinum ssp. gussoneanum (H. geniculatum All.)]-T. aestivum alloplasmic recombinant line and the wheat-rye substitution lines 1R (1A), 1R (1D), and 3R(3B). Chromosomes 1R and 3R of rye cultivar Onokhoiskaya proved to affect the expression of polyembryony in the hybrid combinations that involved the alloplasmic recombinant lines of common wheat as maternal genotypes. Based on this finding, polyembryony was regarded as a phenotypic expression of nuclear-cytoplasmic interactions where an important role is played by rye chromosomes 1R and 3R and the H. vulgare cytoplasm. Consideration is given to the association between the effect of rye chromosomes 1R and 3R on polyembryony in the [(Hordeum)-T. aestivum x wheat-rye substitution lines] hybrid combinations and their stimulating effect on the development on angrogenic embryoids in isolated anther cultures of the wheat-rye substitution lines.     

The Effect of Parental Genotypes of Rye Lines on the Development of Quantitative Traits in Primary Octoploid Triticale: Plant Height

When breeding the primary spring octoploid triticale derived from crosses of various inbred rye lines to wheat Chinese Spring, the effects of the rye genotype and growth conditions on the plant height and proportion of the first, second, and final (pedicle) internodes to the entire stem length were studied. Two triticale groups were examined: homozygotes for the dominant (Ddw1) and recessive (ddw1) alleles of the gene responsible for short stem in rye. In the short stem triticale lines carrying the Ddw1 alleles, the plants were 20 cm shorter on average than those in the ddw1-carrying lines, and the distribution of the two triticale groups overlapped significantly. In both groups, the lines significantly differing in plant height could be differentiated, because of allelic diversity of the additional genes controlling this trait along with the Ddw gene. In most triticale lines, especially in theDdw1-carrying ones, the plant height was much reduced under unfavorable growth conditions. At the same time, a short-stem line was isolated, which is characterized by ecological plasticity, like the maternal wheat cultivar. In the triticale studied, the stem structure depended on the short-stem rye genotype. The two-year study showed that in the triticale carrying the dominant allele of this gene, the first internode is significantly extended, whereas the upper (pedicle) internode is reduced, which increases plant lodging resistance. The differences revealed between the rye lines as well as their effect on the quantitative triticale traits are discussed in view of a variant of the hybridological analysis, which had been previously proposed for identification and mapping of the correspondent rye genes.     

Segregation distortion caused by weak hybrid necrosis in recombinant inbred lines of common wheat

Segregation distortion of molecular markers is closely related to hybrid incompatibility in progeny from intraspecific crosses. Recent reports in higher plants have demonstrated that hybrid sterility results in segregation distortion at the causal gene regions in progeny of intraspecific crosses. Ne1 and Ne2 complementary loci are known to control hybrid necrosis in intraspecific crosses of common wheat cultivars. Here, we examine the effect of a weak necrosis allele Ne1 ^w on the segregation ratio of molecular markers in recombinant inbred lines (RILs) of common wheat. Some RILs showed accelerated cell death in the leaves at the heading stage due to the epistatic interaction between two quantitative trait loci (QTL) on chromosomes 5B and 2B. Chromosomal localization of these QTL corresponding to Ne1 ^w and Ne2 showed distorted segregation ratios of assigned markers having oppositely biased direction. Although the Ne1 ^w and Ne2 interaction had no obvious effect on seed fertility, Ne1 ^w reduced completion of grain development under the Ne2-homozygous background. This reduction might be one of causes that induces segregation distortion in the 5B and 2B chromosomal regions of RILs. The present study demonstrated that weak hybrid necrosis has limited phenotypic effects; it causes segregation distortion in progeny from intraspecific crosses.