Analysis of Linkage Between Biochemical and Morphological Markers of Rye Chromosomes 1R, 2R,and 5R and Mutations of Self-Fertility at the Main Incompatibility Loci

In F₂ hybrids between self-sterile plants of the Volkhova cultivar and self-fertile lines with established self-fertility mutations (sf mutations) at the major incompatibility loci S (1R), Z (2R), and T (5R), the effect of sf mutations on the inheritance of secalin-encoding, isozyme, and morphological markers located on the same chromosomes was investigated. Linkage between loci Prx7 and Sand locus Sec3 coding for high-molecular-weight secalins on chromosome 1R was shown for the first time. The frequency of recombination between Prx7andSec3and between S and Sec3was 29.1 ± 4.8% and 30.9 ± 7.0%, respectively. Independent inheritance of locus Z and isozyme markers of chromosome 2R, Est3/5 and -Glu, from locus Sec2 encoding 75-kDa -secalins was shown; in hybrids, the recombination frequency between Est3/5 and locus Z varied from 19.2 ± 8.1 to 50%. Independent inheritance of morphological (Ddw and Hs) and isozyme markers (Est4, Est6/9,and Aco2) of chromosome 5R from locus Tlocated on the same chromosome was demonstrated.     

Genetic studies of self-fertility in rye (Secale cereale L.). 2. The search for isozyme marker genes linked to self-incompatibility loci

The segregation of several isozyme marker genes has been studied in F₂ inbred families from hybrids between self-sterile and five self-fertile inbred lines (nos. 2, 3, 4, 5, and 8) as well as from interline hybrids. Self-pollination of F₁ hybrids between self-sterile forms and lines 5 and 8 gave an F₂ segregation ratio of 1 heterozygote:1 homozygote for the gene Prx7 (chromosome 1R) against the allele from the line. This is interpreted as a result of tight linkage of the Prx7 gene with the S1 gene in chromosome 1R (recombination at a level of 0–1%). The self-pollination of such hybrids with lines 2,3 and 4 gave normal segregation for the Prx7 gene (1:2:1). This means that these lines carry a self-fertility allele which is not on chromosome 1R. Interline hybrids 5×2, 5×3 and 5×4 had self-fertility alleles for the two S genes and in inbred F₂ progenies gave the expected deviating segregation for the Prx7 gene in a ratio of 2:3:1. The segregation of interline hybrid 5×8 was normal, 1:2:1, as expected. Highly-deviating segregation in an inbred F₂ family of a hybrid with line 5 has also been obtained for another gene from chromosome 1R — Pgi2 (recombination with the S1 locus of 16.7%). By using the same method it has been estimated that line 4 has a self-fertility allele of the S2 locus from chromosome 2R and that the genes -Glu and Est4/11 are linked with it (recombination 16.7% and 17.5–20% respectively). Lines 2 and 3 have a self-fertility allele of the S5 locus from chromosome 5R which is linked with the Est5-7 gene complex (recombination at a level of 28.8–36.0%).     

Mapping the nucleolus organizer region,seed protein loci and isozyme loci on chromosome 1R in rye

Summary The nucleolus organizer region located on the short arm of chromosome 1R of rye consists of a large cluster of genes that code for ribosomal RNA (designated the Nor-R1 locus). The genes in the cluster are separated by spacer regions which can vary in length in different rye lines. Differences in the spacer regions were scored in two families of F₂ progeny. Segregation also occurred, in one or both of the families, at two seed protein loci and at two isozyme loci also located on chromosome 1R. The seed protein loci were identified as the Sec 1 locus controlling -secalins located on the short arm of chromosome 1R and the Sec 3 locus controlling high-molecular-weight secalins located on the long arm of 1R. The two isozyme loci were the Gpi-R1 locus controlling glucose-phosphate isomerase isozymes and the Pgd 2 locus controlling phosphogluconate dehydrogenase isozymes. The data indicated linkage between all five loci and map distances were calculated. The results indicate a gene order: Pgd 2 ... Sec 3 ... [centromere] ... Nor-R1 ... Gpi-R1 ... Sec 1. Evidence was obtained that rye possesses a minor 5S RNA locus (chromosome location unknown) in addition to the major 5S RNA locus previously shown to be located on the short arm of chromosome 1R.     

Genetic effects of chromosomal rearrangements in Chinese hamster ovary cells: expression and chromosomal assignment of TK, GALK, ACP1, ADA, and ITPA loci.

Polyethylene glycol-mediated fusion of Chinese hamster ovary (CHO) cells with mouse Cl1D cells produced interspecific somatic cell hybrids which slowly segregated CHO chromosomes. Cytogenetic and isozyme analysis of HAT- and bromodeoxyuridine-selected hybrid subclones and of members of a hybrid clone panel retaining different combinations of CHO chromosomes enabled provisional assignments of the following enzyme loci to CHO chromosomes: TK, GALK, and ACP1 to chromosome 7; TK and GALK to chromosome Z13; ACP1, ADA, and ITPA to chromosome Z8; and ADA and ITPA to chromosome Z9. These genetic markers reflect the origin of each of these Z group chromosomes and indicate the functional activity of alleles located on rearranged chromosomes. Identification of diploid electrophoretic shift mutations for ADA and ITPA was consistent with those observations. Assignment of the functional TK locus in TK+/- CHO-AT3-2 cells indicated that gene deletion may be responsible for TK hemizygosity in this subline.     

不同类型杂交水稻组合及其亲本苗期同工酶位点分析

对6个杂交水稻组合及其亲本苗期14种酶的同工酶进行检测,并对其中较为清晰稳定的酶带进行了位点分析,以探讨杂种优势遗传基础。共获得了28个位点的相关资料,包括每个位点的性质、表型多态性和每个位点所编码的酶活性及酶带平均迁移率。所有供试材料在其中25个位点上均未表现出多态性,只有部分组合与其亲本在Est-4、Amy-1、Aat-2等3个位点上存在多态性,其中有81．8％的差异组合其酶带表现出共显性特征。对共显性与杂种优势的关系进行了探讨。     

栗种间杂交系的同工酶基因与形态标记的遗传连锁关系研究

探讨了美州栗（ＣａｓｔａｎｅａｄｅｎｔａｔａＢｏｒｋｈ）与中国栗（Ｃ．ｍｏｌｌｉｓｓｉｍａＢｌｕｍｅ）种间杂交的１个Ｆ２代和２个回交一代家系中８个同工酶基因,Ａｃｐ３、Ｅｓｔ５、Ｐｒｘ１、Ｐｒｘ２、Ｐｒｘ３,Ｍｅ和Ａｄｈ与２个形态标记：Ｉｎｈ和Ｔｗｈ的连锁遗传关系,研究结果表明,在所有家系中,Ｉｎｈ与Ｐｒｘ１和Ｅｓｔ５具有一致的连锁关系,其基因的连锁顺序为Ｉｎｙ－Ｐｒｘ１－Ｅｓｔ５。还发现有参试的     

Linkage of genes, controlling morphological signs with isozyme markers of rye chromosomes

Data on linkage of 12 rye genes controlling morphological traits (el, Vs, ln, w, np, ct2, Hs, Ddw, cb, mn, vil, mp) with one or several isozyme markers of individual rye chromosomes (2R-7R) are presented. Linkage of the following gene pairs was established: chromosome 2R: Est3/5-el, el-beta-Glu, Sod2-el, Sod2-Vs; chromosome 3R: ln-Got4; chromosome 4R: w-Got1, np-Got1; chromosome 5R: Est4-ct2, Est6/9-ct2, ct2-Est2, ct2-Aco2, Est2-Hs, Aco2-Hs, Est2-Ddw, Aco2-Ddw; chromosome 6R: Lap2-cb, cb-Aco1, Est10-mn; chromosome 7R: Acph2/3-vi1, Got2-vi1, mp-Acph2/3. The reasons for mapping a very small number of genes in rye in spite of high intraspecific variability of this species are discussed. An approach is suggested to improve this situation by simultaneous identification and mapping of all diverse spontaneous mutations maintained in heterozygous state in various rye cultivars.     

Distorted segregation of the esterase isozyme genotypes in barley (Hordeum vulgare L.)

Distorted segregation of the esterase isozyme genotypes was observed in F2 population which was produced from a cross combination between 'Ko A' and 'Mokusekko 3' of barley varieties. After examining the segregation of esterase isozyme genotypes in B1F1 hybrids derived from the reciprocal backcrosses, it was made clear that the distorted segregation was caused by certation between pollens of different genotypes, independent of the female genotypes. Furthermore, the certation was controlled by a newly designated gene, Ga2, at the locus which was linked with the multiocus, Est1, Est2 and Est4, for esterase isozymes at the long arm of chromosome 3. The distorted segregation ratios did not significantly vary among F2 populations derived from F1 hybrids of the same combination which were grown in different years. Mechanically mixed pollens of the parents, however, could not induce the certation.     

Mapping of two new markers within the smallest interval harboring the spinal muscular atrophy locus by family and radiation hybrid analysis

The locus responsible for the childhood-onset proximal spinal muscular atrophies (SMA) has recently been mapped to an area of 2–3 Mb in the region q12–13.3 of chromosome 5. We have used a series of radiation hybrids (RHs) containing distinct parts of the SMA region as defined by reference markers. A cosmid library was constructed from one RH. Thirteen clones were isolated and five of these were mapped within the SMA region. Both RH mapping and fluorescence in situ hybridization analysis showed that two clones map in the region between loci D5S125 and D5S351. One of the cosmids contains expressed sequences. Polymorphic dinucleotide repeats were identified in both clones and used for segregation analysis of key recombinant SMA families. One recombination between the SMA locus and the new marker 9Ic (D5S685) indicates that 9Ic is probably the closest distal marker. The absence of recombination between the SMA locus and marker Fc (D5S684) suggests that Fc is located close to the disease gene. These new loci should refine linkage analysis in SMA family studies and may facilitate the isolation of the disease gene.     

Discovery and mapping of wheat Ph1 suppressors

Pairing between wheat (Triticum turgidum and T. aestivum) homeologous chromosomes is prevented by the expression of the Ph1 locus on the long arm of chromosome 5B. The genome of Aegilops speltoides suppresses Ph1 expression in wheat x Ae. speltoides hybrids. Suppressors with major effects were mapped as Mendelian loci on the long arms of Ae. speltoides chromosomes 3S and 7S. The chromosome 3S locus was designated Su1-Ph1 and the chromosome 7S locus was designated Su2-Ph1. A QTL with a minor effect was mapped on the short arm of chromosome 5S and was designated QPh.ucd-5S. The expression of Su1-Ph1 and Su2-Ph1 increased homeologous chromosome pairing in T. aestivum x Ae. speltoides hybrids by 8.4 and 5.8 chiasmata/cell, respectively. Su1-Ph1 was completely epistatic to Su2-Ph1, and the two genes acting together increased homeologous chromosome pairing in T. aestivum x Ae. speltoides hybrids to the same level as Su1-Ph1 acting alone. QPh.ucd-5S expression increased homeologous chromosome pairing by 1.6 chiasmata/cell in T. aestivum x Ae. speltoides hybrids and was additive to the expression of Su2-Ph1. It is hypothesized that the products of Su1-Ph1 and Su2-Ph1 affect pairing between homeologous chromosomes by regulating the expression of Ph1 but the product of QPh.ucd-5S may primarily regulate recombination between homologous chromosomes.     

ISSR and isozyme characterization of androgenetic dihaploids reveals tetrasomic inheritance in tetraploid somatic hybrids between Solanum melongena and Solanum aethiopicum group Gilo

Gene exchanges between Solanum melongena and its allied relative Solanum aethiopicum are a crucial prerequisite for introgression of useful traits from the allied species into the cultivated eggplant. In order to evaluate the extent of genetic recombination between the 2 species, biochemical and molecular markers were employed. A dihaploid population obtained through anther culture of the corresponding tetraploid somatic hybrids was genetically analyzed. The extent of disomic/tetrasomic inheritance and segregation ratios of 3 isozyme systems and intersimple sequence repeat (ISSR) markers were evaluated. The dihaploids, being derived from microspores, allowed for simple, complete, and accurate analyses. The segregation of 280 ISSR markers (110 aethiopicum-specific, 104 melongena-specific, and 66 monomorphic) were evaluated in 71 dihaploids. According to the genetic constitution (simplex/duplex/triplex), almost 64% of the fragments revealed the tetrasomic and/or disomic inheritance. With regard to the assigned species-specific fragments, 68% and 4% were unambiguously the result of tetrasomic and disomic inheritance, respectively. Twenty-four of the 66 monomorphic ISSRs were inherited according to random chromatid segregation. The phenotypes of glucose-6-phosphate dehydrogenase (G-6-PDH), 6-phosphogluconate dehydrogenase (6-PGDH), and shikimate dehydrogenase (SKDH) were studied in 70 dihaploids and inferences were made about the allelic state of their 5 loci. The isozyme markers segregated in the dihaploids in a distorted manner, their segregations did not fit in with any of the expected segregation ratios. However, tetrasomic inheritance might be suggested for G-6-PDH 2 and SKDH 1 loci. Our results demonstrated that gene exchanges occurred readily in the somatic hybrids between S. melongena and S. aethiopicum gr. Gilo.     

Polymorphism at the esterase isozyme locus Est10 associated with phylogenetic differentiation in rice

A new esterase isozyme locus, Est10, with 6 alleles including the null form, has been found in rice by using polyacrylamide gel electrophoresis. Thirty F(2) populations of all possible combinations between 5 different band morphs were studied. The segregation pattern indicated that bands 1, 2, 3, 4, and the null form (0) were allelic with each other. The alleles of Est10 were distributed at different frequencies among different varietal groups of rice and also between cultivated rice and its wild relatives (Oryza rufipogon Griff.). Alleles 1 and 2 were frequently found in Japonica and Indica types, respectively. Allele 3 showed a high frequency in Aus and Boro, both Indica types cultivated in South Asia. Allele 4 was frequent in wild rice O. rufipogon. Judging from the linkage between Est10 and RFLP marker RG220 and isozyme marker Est5, Est10 is located on chromosome 1. The importance of this locus in evolutionary studies of rice is discussed.     

Inheritance and linkage of isozymes in sweet cherry (Prunus avium L.)

Eight polymorphic isozyme loci, 6PGD, G6PD, MDH, PGM, SKDH, FDP, GOT and IDH, in sweet cherry where found to be in one linkage group, with a ninth isozyme locus, GPI, being in another linkage group on a different chromosome. Isozymes were also linked to the incompatibility S locus and this explained the disturbed segregation ratios observed in the first generation from controlled hybridisations between different sweet cherry cultivars. Analysis revealed close linkage between the isozyme and S loci. The results supported a pre-existing theory that the S gene in cherry consists of three linked segments each coding for a different function. Progeny derived from selfing of Stella, the self-fertile cherry cultivar, also showed disturbed segregation ratios and an absence of homozygotes for the isozyme loci assayed. This demonstrated that codominant inheritance of the S alleles had not been effected by the self-fertile mutation.     

Mapping of three self-fertility mutations in rye (Secale cereale L.) using RFLP, isozyme and morphological markers

 Three mutations determining self-fertility at the S, Z and S5 self-incompatibility loci on chromosomes 1R, 2R and 5R of rye, respectively, were mapped using three different F₂ populations. There was a close linkage of one isozyme and four RFLP markers, and no recombinant plants were detected. These markers are Prx7, Xiag249 and Xpsr634 for the S locus (1R), Xbcd266 for the Z locus (2R) and Xpsr100 for the S5 locus (5R). Linkage data for markers associated to the self-fertility mutations at the S, Z and S5 loci were calculated and compared with genetic maps computed by MAPMAKER multipoint analysis. Received: 8 October 1997 / Acepted: 26 November 1997     

Resolution of the two loci for autosomal dominant aniridia, AN1 and AN2, to a single locus on chromosome 11p13.

Two distinct loci have been proposed for aniridia; AN1 for autosomal dominant aniridia on chromosome 2p and AN2 for the aniridia in the WAGR contiguous gene syndrome on chromosome 11p13. In this report, the kindred segregating for autosomal dominant aniridia, which suggested linkage to acid phosphatase-1 (ACP1) and led to the assignment of the AN1 locus on chromosome 2p, has been updated and expanded. Linkage analysis between the aniridia phenotype and ACP1 does not support the original linkage results, excluding linkage up to theta = 0.17 with Z = -2. Tests for linkage to other chromosome 2p markers. APOB, D2S71, D2S5, and D2S1, also excluded linkage to aniridia. Markers that have been isolated from the chromosome 11p13 region were then analyzed in this aniridia family. Two RFLPs at the D11S323 locus give significant evidence for linkage. The PvuII polymorphism detected by probe p5S1.6 detects no recombinants, with a maximum lod score of Z = 6.97 at theta = 0.00. The HaeIII polymorphism detected by the probe p5BE1.2 gives a maximum lod score of Z = 2.57 at theta = 0.00. Locus D11S325 gives a lod score of Z = 1.53 at theta = 0.00. These data suggest that a locus for aniridia (AN1) on chromosome 2p has been misassigned and that this autosomal dominant aniridia family is segregating for an aniridia mutation linked to markers in the 11p13 region.     

Inheritance of enzymes and blood proteins in the leopard frog,Rana pipiens: Three linkage groups established

Individuals from natural populations of the leopard frog, Rana pipiens, were analyzed for electrophoretic differences in blood proteins and enzymes from an amputated digit. The proteins examined represent products of 72 loci. Presumptive heterozygotes at multiple loci were selected for experimental crosses. Mendelian inheritance of 18 protein variations were demonstrated in the offspring. Tests for linkage or independent assortment were performed for 75 locus pairs. Three linkage groups were established. Linkage group 1 contains two loci, aconitase-1 (Acon1) and serum albumin (Alb), with a 19% recombination frequency between them. Linkage group 2 contains four loci, glyoxalase (Gly), acid phosphatase-1 (Ap1), acid phosphatase-2 (AP2), and esterase-5 (Est5). The data show the relationships Gly-21.1%-AP1-0%-AP2-6.3%-Est5, and Gly-25.6%-Est5. Linkage group 3 consists of four closely linked esterase loci. The data, Est1-5.1%-Est6, Est6-1.8%-Est10-1.9%-Est4 and Est6-3.0%-Est4, do not establish a complete order but suggest that Est10 is between Est4 and Est6. These results, with data demonstrating apparent independent assortment of 67 other locus pairs, provide a foundation for establishing the frog genetic map.The project was supported by Grant No. RR-00572 from the Division of Research Resources, National Institutes of Health. This paper is contribution No. C-87 from the Amphibian Facility, George W. Nace, Director.     

Linkage between an incompatibility locus and a peroxidase isozyme locus (Prx 7) in rye

Summary Under controlled growth chamber conditions of 30 °C, seed set after selfing is possible in normally self-incompatible rye plants. Within selfed progenies produced by this method, plants homozygous at the peroxidase isozyme locus Prx 7 were crossed to heterozygous individuals. Segregation at the Prx 7 locus in progenies of these crosses provides clear evidence of a close linkage between Prx 7 and one of the two incompatibility loci in rye. A recombination fraction in the range of 0–2% was calculated from the segregation data. In rye, Prx 7 is linked with a phosphoglucoisomerase locus (Pgi). The similarity between the observations in Secale cereale and those made in Lolium perenne is discussed.     

Mapping Three Classical Isozyme Loci in Tetrahymena: Meiotic Linkage of Esta to the Chxa Linkage Group

We demonstrate a reliable method for mapping conventional loci and obtaining meiotic linkage data for the ciliated protozoan Tetrahymena thermophila. By coupling nullisomic deletion mapping with meiotic linkage mapping, loci known to be located on a particular chromosome or chromosome arm can be tested for recombination. This approach has been used to map three isozyme loci, EstA (Esterase A), EstB (Esterase B), and AcpA (Acid Phosphatase A), with respect to the ChxA locus (cycloheximide resistance) and 11 RAPDs (randomly amplified polymorphic DNAs). To assign isozyme loci to chromosomes, clones of inbred strains C3 or C2 were crossed to inbred strain B nullisomics. EstA, EstB and AcpA were mapped to chromosomes 1R, 3L and 3R, respectively. To test EstA and AcpA for linkage to known RAPD loci on their respective chromosomes, a panel of Round II (genomic exclusion) segregants from a B/C3 heterozygote was used. Using the MAPMAKER program, EstA was assigned to the ChxA linkage group on chromosome 1R, and a detailed map was constructed that includes 10 RAPDs. AcpA (on 3R), while unlinked to all the RAPDs assigned to chromosome 3 by nullisomic mapping, does show linkage to a RAPD not yet assignable to chromosomes by nullisomic mapping.     

Segregation of recessive phenotypes in somatic cell hybrids: role of mitotic recombination, gene inactivation, and chromosome nondisjunction.

Somatic cell hybrids heterozygous at the emetine resistance locus (emtr/emt+) or the chromate resistance locus (chrr/chr+) are known to segregate the recessive drug resistance phenotype at high frequency. We have examined mechanisms of segregation in Chinese hamster cell hybrids heterozygous at these two loci, both of which map to the long arm of Chinese hamster chromosome 2. To follow the fate of chromosomal arms through the segregation process, our hybrids were also heterozygous at the mtx (methotrexate resistance) locus on the short arm of chromosome 2 and carried cytogenetically marked chromosomes with either a short-arm deletion (2p-) or a long-arm addition (2q+). Karyotype and phenotype analysis of emetine- or chromate-resistant segregants from such hybrids allowed us to distinguish four potential segregation mechanisms: (i) loss of the emt+- or chr+-bearing chromosome; (ii) mitotic recombination between the centromere and the emt or chr loci, giving rise to homozygous resistant segregants; (iii) inactivation of the emt+ or chr+ alleles; and (iv) loss of the emt+- or chr+-bearing chromosome with duplication of the homologous chromosome carrying the emtr or chrr allele. Of 48 independent segregants examined, only 9 (20%) arose by simple chromosome loss. Two segregants (4%) were consistent with a gene inactivation mechanism, but because of their rarity, other mechanisms such as mutation or submicroscopic deletion could not be excluded. Twenty-one segregants (44%) arose by either mitotic recombination or chromosome loss and duplication; the two mechanisms were not distinguishable in that experiment. Finally, in hybrids allowing these two mechanisms to be distinguished, 15 segregants (31%) arose by chromosome loss and duplication, and none arose by mitotic recombination.     

Genetic and in vitro molecular hybridization of malate dehydrogenase isozymes in interspecific bass (Micropterus) hybrids

Supernatant malate dehydrogenase (MDH) isozymes (as visualized by starch gel electrophoresis) are encoded by two distinct gene loci in both the largemouth and smallmouth bass. When an interspecific F₁ hybrid is formed between these two fish, a unique MDH isozyme is generated. The results of freeze-thaw molecular hybridization (which is the first application of this technique to MDH) indicate that this unique isozyme in the F₁ hybrid is a heterodimer composed of one subunit of each parental type. The F₁ hybrids produced F₂ hybrids which in turn formed the F₃ hybrid population. The inheritance of alleles at the MDH-B locus is consistent with a single Mendelian autosomal locus. Furthermore, there is no evidence of linkage between the lactate dehydrogenase-E locus and the MDH-B locus.