Genetic Analysis of Inheritance ofmei8, sy1, and sy10Meiotic Mutations in Rye Secale cereale L.

It is shown that mutations mei8 (irregular condensation and fragmentation of meiotic chromosomes),sy1(asynapsis), and sy10 (heterologous synapsis) of rye Secale cereale L. are nonallelic. In double mutants mei8 sy1 and mei8 sy10 both mutations are expressed simultaneously and independently of each other. A study of joint inheritance of mutations sy1 and sy10 revealed their interaction by means of recessive epistasis: the double mutants sy1sy10 has the sy10 phenotype. This means that the sy10 gene controls an earlier stage of synapsis in meiotic prophase than the sy1 gene. Mutation mei8is inherited independently of sy1 but it is linked to sy10 (recombination frequency 26.8 ± 3.58%).     

Genetic Control of Chromosome Synapsis at Meiosis in Rye Secale cereale L.: The sy19 Gene Controlling Heterologous Synapsis

Analysis of manifestation and inheritance of a new mutation inducing irregular synapsis in rye showed that abnormal phenotype is determined by a recessive allele of the sy19 gene. In the homozygotes for this mutation, even at the light microscopic level, abnormal formation of bivalents is already observed at pachytene–diakinesis. At metaphase I, the univalent frequency varies from 0 to 14; in a few cells, multivalent associations of chromosomes, which are not clearly oriented in the spindle, are detected. Electron microscopy of synaptonemal complexes revealed both homologous and heterologous synapsis in homozygotes for sy19, namely partial loss of the ability to stringent homology search. Analysis of joint inheritance of sy19 and asynaptic sy1 mutations showed that they are nonallelic, inherited independently, and interact by recessive epistasis. The phenotype of doublesy1sy19 mutants indicates that thesy19 gene conditioning heterologous synapsis operates at meiosis later than the synaptic gene sy1. The epistatic group of mutations, sy9 > sy1 > sy19 and sy3, was determined.     

Genetic Analysis of Mutation sy2 which Causes Nonhomologous Meiotic Synapsis in Chromosomes of Diploid Rye Secale cereale L.

Partially nonhomologous (heterologous) synapsis of meiotic chromosomes in a spontaneous desynaptic mutant form of rye is determined by two recessive genes, sy2a and sy2b, that have independent expression and inheritance. The third gene, dominant inhibitor suppressing the mutant phenotype, has been revealed in hybrid combinations between sy2 mutants and lines segregating other meiotic mutants: sy10 (heterologous synapsis), sy1, and sy9(asynapsis). All three genes determining desynapsis (sy2a, sy2b, and I) were shown to be nonallelic to monogenic mutations sy10, sy1, and sy9, inherited independently of them and expressed at later stages of prophase I than the sy10 gene. The possibility of modifying monogenic segregation of mutation sy2 by gametophyte selection for a locus linked to the gene expressed as sy2 at particular frequencies of recombination between this gene and selected locus is discussed.     

Genetic analysis of inheritance of mei8, sy1 and sy10 mutations,disrupting the correct meiosis in the rye Secale cereale L

It is shown that mutations mei8 (irregular condensation and fragmentation of meiotic chromosomes), sy1 (asynapsis), and sy10 (heterologous synapsis) of rye Secale cereal are nonallelic. In double mutants mei8 sy1 and mei8 sy10 both mutations are expressed simultaneously and independently of each other. A study of joint inheritance of mutations sy1 and sy10 revealed their interaction by means of recessive epistasis: the double mutants has the sy10 phenotype. This means that the sy10 gene controls an earlier stage of synapsis in meiotic prophase than the sy1 gene. Mutation mei8 is inherited independently of sy1 but it is linked to sy10 (recombination frequency 26.8 +/- 3.58%).     

Meiotic mutations in rye Secale cereale L

Spontaneous meiotic mutations of winter rye Secale cereale L. (2n = 14) were revealed in inbred F2 progenies, which were obtained by self-pollination of F1 hybrids resulting from crosses of individual plants of cultivar Vyatka or weedy rye with plants of self-fertile inbred lines. The mutations cause partial or complete sterility, and are maintained in heterozygote condition. Six types of mutations were distinguished as the result of cytological analysis of meiosis and genetic analysis. (1) Plants with nonallelic asynaptic mutations sy1 and sy9 lacked bivalents in 96.8 and 67.0% metaphase I cells, respectively, formed only axial elements but not the mature synaptonemal complex (SC), and had defects in telomere clustering in early prophase I. (2) Weak asynaptic mutant sy3 showed incomplete synapsis at the start of SC degradation at diplotene and lower chiasma number; yet only 2% meiocytes lacked bivalents in MI. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19 caused nonhomologous synapsis; i.e., a varying number of univalents and occasional multivalents were observed in MI, which was preceded by switches of pairing partners and fold-back synapsis at mid-prophase I. (4) Mutation mei6 led to the formation of protrusions and minor branched structures of the SC lateral elements. (5) Allelic mutations mei8 and mei8-10 caused irregular chromatin condensation along the chromosome length in prophase I, which was accompanied by chromosome sticking and fragmentation in MI. (6) Allelic mutations mei5 and mei10 determined chromosome supercondensation, caused the disturbance of meiotic spindle assembly, arrested meiosis at various stages but did not affect formation of the pollen wall, thus arrested meiocytes got covered with the pollen wall. Analysis of double mutants revealed recessive epistatic interactions for some mutations; the epistatic group was sy9 > sy1 > sy3 > sy19. This reflects the sequence of meiotic events controlled by the corresponding genes. The expression of sy2 and sy19 proved to be modified by additional genes. Most meiotic mutations found in rye have analogs in other plants.     

The relation between synapsis and chiasma formation in Rhoeo spathacea

Rhoeo spathacea (Schwartz) Stearn (2n=12) is a complete translocation heterozygote in which a ring of 12 chromosomes is often observed from diakinesis through metaphase I. A new hypotonie bursting technique on primary microsporocytes made possible visualization of the complete pattern of pachytene synapsis in R. spathacea by light and electron microscopy. Analysis showed that most of the length of a chromosome never synapsed and that relational twisting of the lateral elements was prominent in the distal segments of most synaptonemal complexes (SCs). The most common situation was 12 relatively short SCs representing the 24 synapsed terminal segments of the 12 chromosomes; 11 and 10 SCs were also observed but less frequently, indicating one and two synaptic failures, respectively. Similarly, in diakinesis a ring of 12 chromosomes was most common, but a chain of 12 chromosomes and two chains of chromosomes could also occur. These situations indicated zero, one, and two chiasma failures, respectively. The frequencies of synaptic failure (0.021) and of chiasma failure (0.022) were essentially the same as were the distributions of cells with different numbers of synaptic and chiasma failures. These comparisons indicate that if synapsis occurs between two distal segments of the chromosomes of R. spathacea, a chiasma will almost certainly follow. A diagram to illustrate the arrangement of mid-pachytene chromosomes and a model to explain the origin of relational twisting of lateral elements in SCs are presented.     

Impairment of homologous chromosome synapsis in meiosis in rye <Emphasis Type="Italic">Secale cereale</Emphasis> L. Caused by a recessive mutation of the <Emphasis Type="Italic">sy18</Emphasis> gene

Expression and inheritance of the sy18 mutation causing impairment of synapsis homology were studied. It was established that the abnormal phenotype is determined by a recessive allele of the sy18 gene. Univalents and multivalents are observed in homozygotes for this mutant allele. According to the electron microscopic analysis of synaptonemal complexes in mutants, homologous synapsis occurs together with nonhomologous synapsis. The sy18 gene was found to have no allelism with asynaptic genes sy1 and sy9 and with genes sy10 and sy19 causing, like sy18, disturbances in synapsis homology.     

Genetic collection of meiotic mutants of rye Secale cereale L

Genetic collection of meiotic mutants of winter rye Secale cereale L. (2n = 14) was created. Mutations were detected in inbred F2 generations after self-fertilization of the F1 hybrids, obtained by individual crossing of rye plants (cultivar Vyatka) or weedy rye with plants from autofertile lines. The mutations cause partial or complete plant sterility and are maintained in collection in a heterozygous state. Genetic analysis accompanied by cytogenetic study of meiosis has revealed six mutation types. (1) Nonallelic asynaptic mutations sy1 and sy9 caused the formation of only axial chromosome elements in prophase and anaphase. The synaptonemal complexes (SCs) were absent, the formation of the chromosome "bouquet" was impaired, and all chromosomes were univalent in meiotic metaphase I in 96% (sy1) and 67% (sy2) of cells. (2) Weak asynaptic mutation sy3, which hindered complete termination of synapsis in prophase II. Subterminal asynaptic segments were always observed in the SC, and at least one pair of univalents was present in metaphase I, but the number of cells with univalents did not exceed 2%. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19, which caused partially nonhomologous synapsis: change in pairing partners and fold-back chromosome synapsis in prophase I. In metaphase I, the number of univalents varied and multivalents were observed. (4) Mutation mei6, which causes the formation of ultrastructural protrusions on the lateral SC elements, gaps and branching of these elements. (5) Allelic mutations mei8 and mei10, which caused irregular chromatin condensation along chromosomes in prophase I, sticking and fragmentation of chromosomes in metaphase I. (6) Allelic mutations mei5 and mei10, which caused chromosome hypercondensation, defects of the division spindle formation, and random arrest of cells at different meiotic stages. However, these mutations did not affect the formation of microspore envelopes even around the cells, whose development was blocked at prophase I. Analysis of cytological pictures of meiosis in double rye mutants reveled epistatic interaction in the mutation series sy9 > sy1 > sy3 > sy19, which reflects the order of switching these genes in the course of meiosis. The expression of genes sy2 and sy19 was shown to be controlled by modifier genes. Most meiotic mutations found in rye have analogs in other plant species.     

Genetic control of chromosomal synapsis in Secale cereale L. rye meiosis: sy19 gene, causing heterologous synapsis

Analysis of manifestation and inheritance of a new mutation inducing irregular synapsis in rye showed that abnormal phenotype is determined by a recessive allele of the sy19 gene. In the homozygotes for this mutation, even at the light microscopic level, abnormal formation of bivalents is already observed at pachytene-diakinesis. At metaphase I, the univalent frequency varies from 0 to 14; in a few cells, multivalent associations of chromosomes, which are not clearly oriented in the spindle, are detected. Electron microscopy of synaptonemal complexes revealed both homologous and heterologous synapsis in homozygotes for sy19, namely partial loss of the ability to stringent homology search. Analysis of joint inheritance of sy19 and asynaptic sy1 mutations showed that they are nonallelic, inherited independently, and interact by recessive epistasis. The phenotype of double sy1sy19 mutants indicates that the sy19 gene conditioning heterologous synapsis operates at meiosis later than the synaptic gene sy1. The epistatic group of mutations, sy9 > sy1 > sy19 and sy3, was determined.     

Maize meiotic mutants with improper or non-homologous synapsis due to problems in pairing or synaptonemal complex formation

During meiotic prophase homologous chromosomes find each other and pair. Then they synapse, as the linear protein core (axial element or lateral element) of each homologous chromosome is joined together by a transverse central element, forming the tripartite synaptonemal complex (SC). Ten uncloned Zea mays mutants in our collection were surveyed by transmission electron microscopy by making silver-stained spreads of SCs to identify mutants with non-homologous synapsis or improper synapsis. To analyse the mutants further, zyp1, the maize orthologue of the Arabidopsis central element component ZYP1 was cloned and an antibody was made against it. Using antibodies against ZYP1 and the lateral element components AFD1 and ASY1, it was found that most mutants form normal SCs but are defective in pairing. The large number of non-homologous synapsis mutants defective in pairing illustrates that synapsis and pairing can be uncoupled. Of the ten mutants studied, only dsy2 undergoes normal homologous chromosome recognition needed for homologous pairing. The dsy2 mutation fails to maintain the SC. ZYP1 elongation is blocked at zygotene, and only dots of ZYP1 are seen at prophase I. Another mutant, mei*N2415 showed incomplete but homologous synapsis and ASY1 and AFD1 have a normal distribution. Although installation of ZYP1 is initiated at zygotene, its progression is slowed down and not completed by pachytene in some cells and ZYP1 is not retained on pachytene chromosomes. The mutants described here are now available through the Maize Genetics Cooperation Stock Center (http://maizecoop.cropsci.uiuc.edu/).     

The pattern of zygotene and pachytene pairing in allotetraploid Aegilops species sharing the D genome

Chromosome pairing behaviour of the allotetraploid Aegilops species sharing the D genome, Ae. crassa (DDMM), Ae. cylindrica (DDCC) and Ae. ventricosa (DDNN), was analyzed by electron microscopy in surfacespread prophase-I nuclei. Synaptonemal-complex analysis at zygotene and pachytene revealed that synapsis in the allotetraploids was mostly between homologous chromosomes, although a few multivalents were also formed. Only homologous bivalents were observed at metaphase-I. It is concluded that the mechanism controlling bivalent formation in these species acts mainly at zygotene by restricting pairing to homologous chromosomes, but also acts at pachytene by preventing chiasma formation in homoeologous associations. These observations are discussed in relation to mechanisms of diploidization of polyploid meiosis.     

Lateral elements inside synaptonemal complex-like polycomplexes in ndt80 mutants of yeast bind DNA

The synaptonemal complex (SC) keeps the synapsed homologous chromosomes together during pachytene in meiotic prophase I. Structures that resemble stacks of SCs, polycomplexes, are sometimes found before or after pachytene. We have investigated ndt80 mutants of yeast, which arrest in pachytene. SCs appear normal in spread chromosome preparations, but are only occasionally found in intact nuclei examined in the electron microscope. Instead, large polycomplexes occur in almost every ndt80 mutant nucleus. Immunoelectron microscopy using DNA antibodies show strong preferential labeling to the lateral element parts of the polycomplexes. In situ hybridization using chromosome-specific probes confirms that the chromosomes in ndt80 mutants are paired and attached to the SCs. Our results suggest that polycomplexes can be involved in binding of chromosomes and possibly also in synapsis.     

Haploid meiosis and its bearing on the phylogeny of pearl millet,Pennisetum typhoides stapf et hubb.

Meiotic chromosome associations in a spontaneously originated haploid plant of pearl millet have been studied and their phyletic significance discussed. Chromosome pairing could be observed at pachytene and diplotene. Out of a total of 285 PMC's studied at diakinesis-metaphase 1, 43 showed one bivalent and 7 had two bivalents per cell. Both rod- and ring-bivalents were observed. Apart from synapsis accompanied by chiasma formation, close associations of univalent chromosomes were observed. Out of 150 cells without true bivalents, 41 showed 1 s-s association and five, 2 s-s pairs per cell. On the basis of the realization of a maximum of two bivalents per cell, as also of a maximum of 2 s-s pairs, it has been inferred that the chromosome complement ofP. typhoides (n=7) has evolved from a basic set ofn=5 chromosomes. Other available evidence supporting this inference is also discussed.     

Relationship between pachytene synapsis, metaphase I associations, and transmission of 2B and 4B chromosomes in rye.

2B rye plants selected for high (H) or low (L) B transmission rate were studied at pachytene and metaphase I of meiosis to determine the relationship between synapsis, bivalents at metaphase I, and B transmission rate. The results show that the 2 B chromosomes (Bs) form bivalents at pachytene in both the H and L lines, whereas the frequency of bivalents at metaphase I is much higher in the H than in the L line. This demonstrates that B transmission is mainly related to the proper association of Bs at metaphase I, as well as that synapsis of the 2 Bs in the L line is normal, but the bivalent is not consolidated by a chiasma in most cases. Crosses were made between 2B plants of the H and L lines in all combinations (H x H, H x L, L x H, and L x L) to obtain 4B plants. Similarly, bivalent formation at pachytene and metaphase I was studied. The results show that 4B plants of the H x H and L x L classes differ significantly at pachytene and metaphase I since the former forms more bivalents. The heterozygous 4 Bs of the H x L and L x H classes show intermediate values. The relation H x H > H x L > L x H > L x L was consistently found for the variables transmission rate, bivalents at pachytene, bivalents at metaphase I, and B mean chiasma frequency. A maternal effect was also found. Our data suggest that there are two separate mechanisms acting upon synapsis and chiasma formation in H and L B chromosomes: (i) there is variable efficiency of the control of synapsis at early stages of meiosis; and (ii) there is variable efficiency of the control of the number of chiasmata.     

Molecular assembly of meiotic proteins Asy1 and Zyp1 and pairing promiscuity in rye (Secale cereale L.) and its synaptic mutant sy10

Assembly of two orthologous proteins associated with meiotic chromosome axes in Arabidopsis thaliana (Asy1 and Zyp1) was studied immunologically at meiotic prophase of meiosis of wild-type rye (Secale cereale) and its synaptic mutant sy10, using antibodies derived from A. thaliana. The temporal and spatial expression of the two proteins were similar in wild-type rye, but with one notable difference. Unlike A. thaliana, in which foci of the transverse filament protein Zyp1 appear to linearize commensurately with synapsis, linear tracts of Asy1 and Zyp1 protein form independently at leptotene and early zygotene of rye and coalign into triple structures resembling synaptonemal complexes (SCs) only at later stages of synapsis. The sy10 mutant used in this study also forms spatially separate linear tracts of Asy1 and Zyp1 proteins at leptotene and early zygotene, and these coalign but do not form regular triple structures at midprophase. Electron microscopy of spread axial elements reveals extensive asynapsis with some exchanges of pairing partners. Indiscriminate SCs support nonhomologous chiasma formation at metaphase I, as revealed by multi-color fluorescence in situ hybridization enabling reliable identification of all the chromosomes of the complement. Scrutiny of chiasmate associations of chromosomes at this stage revealed some specificity in the associations of homologous and nonhomologous chromosomes. Inferences about the nature of synapsis in this mutant were drawn from such observations.     

The Expression of Mutation sy2 Causing Nonhomologous Synapsis in Meiosis of Diploid Rye Secale cereale L.

The cytological expression of spontaneous mutation sy2 isolated from a population of weedy rye was examined. It was demonstrated that the primary defect of meiosis in the mutant plants is nonhomologous synapsis, which occurs simultaneously with the homologous one. An electron microscope study of the synaptonemal complex (SC) at prophase I showed synaptic abnormalities that were manifested as switches of synapting axial elements to the nonhomologous partner and the formation of foldbacks of lateral SC elements. The sy2 mutants are characterized by one to two such events per meiocyte. Nonhomologous synapsis leads to the appearance of univalents at metaphase I (on average 4.16 ± 0.002 per meiocyte) and multivalents (on average 0.12 ± 0.007 per meiocyte). The presence of multivalents in 12% of meiocytes at metaphase I may result from recombination in ectopic regions of homology. It is suggested that the sy2 mutation impairs a component of the system that limits synapsis in meiocytes to only homologous chromosome pairs.     

Expression and Inheritance of a desynaptic phenotype with impaired homologous synapsis in rye

The cytological phenotype was studied in a desynaptic form isolated from a population of rye cultivar Vyatka. The primary defect of desynaptic plants was identified as nonhomologous (heterologous) chromosome synapsis, which was observed by electron microscopy of synaptonemal complexes (SCs) in meiotic prophase I. Synapsis defects involved switches of synapsing axial elements to nonhomologous partners, asynapsis in the switching region, and foldbacks formed by the SC lateral elements. Defective bivalent formation was observed at later stages: the univalent number varied and multivalent chromosome associations were observed in single cells in metaphase I. The desynaptic phenotype was controlled by two recessive genes, sy8a and sy8b, which acted and were inherited independently. In a hybrid combination with line Ku-2/63, the desynaptic phenotype was suppressed by the dominant allele of a third gene for inhibitor I; the segregation in hybrid families corresponded to 57:7.     

Semisterile meiotic mutant <Emphasis Type="Italic">sy11</Emphasis> with heterologous chromosome synapsis in rye <Emphasis Type="Italic">Secale cereale</Emphasis> L.

A study was made of the expression and inheritance of the sy11 mutation, which alters homologous chromosome synapsis in meiotic prophase I of rye. The abnormal phenotype proved to be determined by a recessive allele of a single sy11 gene. Univalents and multivalents were observed in homozygotes for the mutant allele. Analysis of the synaptonemal complex revealed a combination of homologous and nonhomologous synapsis in the mutant. The nonhomologous synapsis frequency significantly decreased in the course of meiotic prophase I in the mutant. The number of chiasmata per bivalent in metaphase I was 1.1 ± 0.01 versus 1.8 ± 0.01 in wild-type plants, and the number of univalents was 2.7 ± 0.06 versus 0.5 ± 0.05 in wild-type plants. As a result, a broad range of abnormalities was observed at subsequent stages of meiosis and led to the formation of defective microspores. Mutant plants were semisterile.     

The maize desynaptic1 mutation disrupts meiotic chromosome synapsis

In most eukaryotes, homologous chromosomes undergo synapsis during the first meiotic prophase. A consequence of mutations that interfere with the fidelity or completeness of synapsis can be failure in the formation or maintenance of bivalents, resulting in univalent formation at diakinesis and production of unbalanced spores or gametes. Such mutations, termed desynaptic mutations, can result in complete or partial sterility. We have examined the effect of the maize desynaptic1-9101 mutation on synapsis, using the nuclear spread technique and electron microscopy to examine microsporocytes ranging from early pachytene until the diplotene stage of prophase I. Throughout the pachytene stage, there was an average of about 10 sites of lateral element divergence (indicating nonhomologous synapsis), and during middle and late pachytene, an average of two and three sites of foldback (intrachromosomal) synapsis, per mutant nucleus, respectively. By the diplotene stage, the number of sites of lateral element divergence had decreased to seven, and there was an average of one foldback synapsis site per nucleus. Lateral element divergence and foldback synapsis were not found in spread pachytene nuclei from normal plants. These results imply that the normal expression of the dsy1 gene is essential for the restriction of chromosome synapsis to homologues. The abundance of nonhomologous synapsis and the persistence of extended stretches of unsynapsed axial elements throughout the pachytene stage of dsy1–9101 meiocytes suggests that this mutation disrupts both the fidelity of homology search and the forward course of the synaptic process. This mutation may identify a maize mismatch repair gene. Dev. Genet. 21:146–159, 1997. © 1997 Wiley-Liss, Inc.     

Comparative analysis of female and male meiosis in three meiotic mutants of tomato.

Female meiosis was analysed in squash preparations of ovules from three meiotic mutants and wild-type plants of tomato. In the completely asynaptic mutant as6, chromosome pairing and chiasma formation were virtually absent in both sexes. In the partially asynaptic mutant asb, with intermediate levels of chromosome pairing at pachytene, there were a higher number of chiasmate chromosome arms in female meiosis than in male meiosis, whereas in the desynaptic mutant as5 there were normal levels of chromosome pairing at pachytene and a similar reduction in chiasma frequency in the two sexes. In wild-type tomato, we found slightly higher numbers of chiasmate chromosome arms in female meiosis than in male meiosis. We propose that the higher female chiasma frequencies in mutant asb and wild-type tomato result from a longer duration of female meiotic prophase. This would allow chromosomes more time to pair and recombine. It is possible that a longer duration of prophase I does not affect mutants as5 and as6, either because the meiotic defect acts before the pairing process begins (in as6) or because it acts at a later stage and involves chiasma maintenance (in as5).