The inheritance of rye seed peroxidases

Summary Genetic analyses were conducted on peroxidase of the embryo and endosperm of seeds of one open pollinated and six inbred lines of cultivated rye (Secale cereale L.), and one line of Secale vavilovii Grossh. The analyses of the individual parts of the S. cereale seed yield a total of 14 peroxidase isozymes. Isozymes m, a, b, c, d, e, f and g (in order from faster to slower migration) were found in the embryo plus scutellum, while isozymes 1, 2, 3, 4, 5 and 6 (also from faster to slower migration) were peculiar of the endosperm. S. vavilovii has isozymes m, c₁, d, e, f and g in its embryo plus scutellum, and isozyme 2 in the endosperm. Segregation data indicated that at least 13 different loci would be controlling the peroxidase of S. cereale. Isozymes a and b are controlled by alleles of the same locus, all the other loci have one active and dominant allele coding for one isozyme, and other null and recessive allele. The estimation of linkage relationships shows that five endosperm loci are linked, and tentative maps are shown. A possible dosage effect and the existence of controlling gene(s) for endosperm isozyme 4 is reported. All these data and the high frequency of null alleles found are discussed in relation to recent reports.     

Studies on theStellaria longipes complex (Caryophyllaceae): Isozyme variability and the relationship betweenStellaria longipes andS. longifolia

Genetic variation within and the relationship betweenStellaria longipes Goldie andS. longifolia Muhl. were studied. Ten enzyme systems were assessed in eight natural populations ofS. longipes (25 loci) and three ofS. longifolia (20 loci) using starch and polyacrylamide gel electrophoresis. Patterns of population differentiation corresponded to geographic distance. There was no evidence that polyploidS. longipes had greater electrophoretic variability than diploidS. longipes. The isozyme data confirmed extensive population differentiation in these species and, within that context, a relatively close relationship betweenS. longipes andS. longifolia. It was postulated that diploids of these two species might be the progenitors of tetraploidS. longipes.     

Ribosomal DNA spacer-length variation inSecale spp. (Poaceae)

Variation in ribosomal DNA spacer length was analysed in 23 populations of 12Secale spp. by restriction enzyme analysis. Digestion of rDNA with Taq I endonuclease enzyme yields spacer fragments that include the subrepeat array and the non-repetitive region downstream of the array. Extensive spacer length variation existed in most species with Taq I fragment lengths ranging from 0.9–3.1 kb. These length variants have been attributed to the differences in number of 134 bp spacer subrepeats within rDNA arrays.S. silvestre was the only species to exhibit a unique spacer length variant of 0.9 kb and this was shown to result from the presence of an extra Taq I site in the spacer. rDNA spacer length frequencies were determined for the species. These frequencies were used to derive phenetic relationships between the species by numerical taxonomic methods. In plots constructed fromGower's distance matrices,S. silvestre appeared well separated from the major cluster consisting of the other species. On the basis of morphological and cytogenetic criteria,S. silvestre is considered the most ancient species. The rDNA data is consistent with this interpretation as it shows a clear differentiation ofS. silvestre from all the other species based on length and nucleotide sequence composition of the spacer region.     

ON THE ORIGIN OF CULTIVATED RYE

From extensive cytological, ecological, and morphological studies, it appears that cultivated rye (Secale cereale L.) originated from weedy products derived from introgression of S. montanum into S. vavilovii. Secale vavilovii appears to have been derived from S. silvestre as a consequence of chromosomal translocations. Secale silvestre was, in turn, derived from S. montanum or a common ancestor. Secale africanum, S. dalmaticum, S. ciliatoglume, and S. kuprijanovii appear to be only slightly modified isolated populations of S. montanum. Populations of S. anatolicum are weedy forms of S. montanum, genealogically and chromosomally distinct from the weedy annual forms from which S. cereale arose.     

CYTOGENETIC AND EVOLUTIONARY STUDIES IN SECALE. I. SOME NEW DATA ON THE ANCESTRY OF S. CEREALE

Khush , G. S., and G. L. Stebbins . (U. California, Davis.) Cytogenetic and evolutionary studies in Secale. I. Some new data on the ancestry of S. cereale. Amer. Jour. Bot. 48(8): 723–730. Illus. 1961.—Cultivated rye, Secale cereale, was crossed with all 4 wild species of the genus. It crosses readily with S. montanum, S. africanum, and S. vavilovii, but crossability of S. cereale and S. silvestre is extremely low. Meiosis in the hybrid S. cereale × silvestre is characterized by high frequency of univalents at metaphase I, reduced chiasma frequency at metaphase I, high frequency of PMC's with unequal separations and laggards at anaphase I and II, high frequency of microspores with micronuclei, and extremely low pollen fertility. These abnormalities occurred less frequently in other hybrids, and consequently the pollen fertility is fairly high: 19.1% in the hybrid S. cereale × vavilovii and 31.3% and 31.8%, respectively, in the hybrids S. cereale × montanum and S. cereale × africanum. An interesting cytogenetic feature of all these hybrids, however, was the formation of a translocation configuration of 6 chromosomes at metaphase I of meiosis. In the hybrid, S. cereale × silvestre, a translocation configuration of 8 chromosomes was observed in a few cells. It is evident, therefore, that the genome of S. cereale differs from the genomes of wild species by 2 translocations. Another small translocation may differentiate S. cereale and S. silvestre, in addition. Thus, on the basis of chromosome arrangements, no particular wild species is most likely to be ancestral to S. cereale. Similarly, Stutz's hypothesis of hybrid origin of S. cereale seems highly improbable. After considering ecological preferences, breeding habits, geographical distribution, morphological and cytological affinities of wild species and cultivated rye, it is concluded that S. cereale evolved from S. montanum as a result of progressive cytological and morphological differentiation and that this differentiation was facilitated, probably, by adaptive superiority of translocation heterozygotes and rearrangement homozygotes.     

Isozyme evidence and the origin ofSenecio vulgaris (Compositae)

An electrophoretic survey of isozyme variation was conducted to test the hypothesis thatSenecio vulgaris L. (2n = 40) is of autotetraploid origin fromS. vernalis Waldst. & Kit. (2n = 20). It was established thatS. vulgaris exhibited fixed heterozygosity at three loci examined, showed disomic inheritance at all polymorphic loci, and contained a gene (Est-1) and an allele (Aat-3b) which were not present in the single population ofS. vernalis surveyed. From this it is concluded thatS. vulgaris is not of autotetraploid origin. Instead, the genetic evidence is in keeping with an allopolyploid origin ofS. vulgaris with the possibility thatS. vernalis acted as one of its two parents.     

Identification of the three chromosomes involved in the translocations which structurally differentiate the genome of Secale cereale L. from those of Secale montanum Guss. and Secale vavilovii Grossh.

A standard tester set of reciprocal translocations in S. cereale and several primary trisomics have been used to identify the chromosomes involved in the natural translocation complexes between S. cereale and S. montanum, and S. cereale and S. vavilovii.Chromosomes I, III and V, in our classification system, were found to be involved in the translocations between cultivated rye, S. cereale, and both the wild species S. montanum and S. vavilovii.     

Giemsa karyotypes and their evolutionary significance inScilla bifolia,S. drunensis,andS. vindobonensis (Liliaceae)

Quantitatively evaluated C-banded and conventional karyograms are presented for the related speciesScilla bifolia subsp.danubialis Speta,S. drunensis (Speta)Speta, andS. vindobonensis Speta. On the basis of banding patterns and karyotype structureS. bifolia subsp.danubialis (2n = 18, 2×) andS. drunensis (2n = 36, 4×) are quite similar, whileS. vindobonensis (2n = 18, 2×) is entirely different. There is a moderate degree of karyotypic variation withinS. bifolia subsp.danubialis andS. drunensis. However, inS. vindobonensis karyotypes and banding patterns are almost completely stable over a geographical range of about 500 km. The present results confirm the recent taxonomic separation ofS. vindobonensis fromS. bifolia, and suggest a considerable phylogenetic distance between these two diploid species. The results are discussed with reference to the morphological characters of the species and their geographical distribution.Evolution ofScilla and Related Genera, II.Dedicated to Univ.-Prof. Dr.Elisabeth Tschermak-Woess on the occasion of her 60th birthday.     

Identification by Giemsa technique of the translocations separating cultivated rye from three wild species of Secale

Complete identification of the translocations involved in evolution of S. vavilovii, S. africanum and S. cereale from S. montanum was attained by meiotic analysis after Giemsa banding technique. Based on the original mitotic karyotype of S. montanum, the different chromosome arms were determined by centromere position and banding pattern of chromosomes for the four species and all of the possible interspecific hybrids. This first consistent scheme of cytogenetic relationships reveals: one translocation each, separating S. montanum from S. vavilovii and S. africanum, two translocations each, separating S. cereale from S. vavilovii and S. africanum, and three translocations each, separating S. cereale from S. montanum and S. africanum, respectively.     

Allozyme divergence among four species ofPodaechmea s. l. and the status ofUrsulaea (Bromeliaceae, Bromelioideae)

The genetic relationships ofAechmea mexicana, A. lueddemanniana, A. macvaughii andA. tuitensis were investigated using starch gel electrophoresis. Eight enzyme systems encoded by ten putative gene loci were resolved in seventeen populations.Nei's (1978) genetic distances were obtained from allelic frequencies and used with UPGMA algorithm. Results indicate that some populations belonging to different species display genetic similarities closer to each other than to some conspecific populations. Our results do not support the proposed genusUrsulaea (Read & Baensch 1994), sinceA. tuitensis was closer toA. lueddemanniana andA. mexicana than toA. macvaughii.     

Electrophoretic variation in esterases and peroxidases of native greek diploidAegilops species(Ae. caudata andAe. comosa,Poaceae)

The esterase and peroxidase patterns in five varieties ofAegilops caudata (genome type C) andAe. comosa (genome type M) were studied in order to elucidate the phylogenetic relationships within and between the two groups. The electrostarch gel electrophoresis technique was applied to extracts of shoot and root of 4-day-old seedlings, and the electropherograms were evaluated by gel densitometer traces. Inspite of considerable isozyme polymorphism, closer relationships in the banding patterns were found between different varieties of a single species than between varieties of the two different species. Esterase and peroxidase patterns of the twoAe. caudata varieties (caudata andpolyathera) are very similar and prove their close phylogenetic relationship. The isozyme affinities withinAe. comosa varieties are illustrated by the seriessubventricosa—biaristata—thessalica. The latter endemic variety has quite a number of characteristic bands and is relatively isolated. Altogether, the electrophoretic data agree well with morphological and cytological similarities (Zhukovsky 1928,Eig 1929,Karataglis 1973, 1975b).     

Allozyme variability and phylogenetic relationships in the cultivated potato (Solanum tuberosum) and related species

Gene frequencies at 13 isozyme loci were determined in three South American taxa of cultivated potatoes [the diploid group (gp.) Stenotomum, the diploid subgroups (subgp.) Goniocalyx, and the tetraploid gp. Andigena ofS. tuberosum], in the diploid weed speciesS. sparsipilum, and in most of the main cultivars now raised in the Northern Hemisphere (the tetraploid gp. Tuberosum ofS. tuberosum). High levels of genetic variability (mean number of alleles per locus, percentage of polymorphic loci, and mean heterozygosity) were detected, being higher in tetraploid potatoes. An equilibrium among the evolutionary factors which increase genetic variability and artificial selection for maximum yield would explain the high uniformity of heterozygosity values we observed in both Andigena (0.36 ± 0.02) and Tuberosum (0.38 ± 0.01) cultivars.—The low value of genetic distance (D = 0.044) between Stenotomum and Goniocalyx does not support the status of species forS. goniocalyx.—In most isozyme loci, the electromorphs of gp. Andigena were a combination of those found in both gp. Stenotomum andS. sparsipilum, suggesting an amphidiploid origin of gp. Andigena from that two diploid taxa. The presence in Andigena of unique electromorphs, which were lacking in both gp. Stenotomum andS. sparsipilum, suggests that other diploid species could be also implied in the origin of tetraploid Andean potatoes. Furthermore, since Andigena were more related to Stenotomum (D = 0.052) than toS. sparsipilum (D = 0.241), the autopolyploidization of Stenotomum individuals and the subsequent hybridization with gp. Andigena may also have occurred. Thus, our study suggests a multiple origin (amphidiploidy, autoploidy, and hybridization at tetraploid level) of gp. Andigena.—Most of the electromorphs of gp. Tuberosum were also found in gp. Andigena; both the direct derivation of that group from the Andean tetraploid potatoes and the repeated introgression provided by breeding programmes could explain this result. However, the allele c of Pgm-B, present in 30 out of 76 Tuberosum cultivars from Northern Hemisphere as well as in 3 Chilean Tuberosum cultivars, lacks in the 258 Andigena genotypes sampled, suggesting that Chilean germplasm could have taken part in the origin of at least the 39% of the potato cultivars from Europe and North America analyzed here.—The distanceWagner procedure provides an estimate of a 30% of heterogeneity in the evolutionary divergence shown by different groups of cultivated potatoes. Diploid groups show a higher (22.5%) evolutionary rate than tetraploids, which can be attributed to both tetrasomic inheritance and facultative autofecundation that exists in Andigena and Tuberosum groups. Thus, artificial selection acting since 10000 years has not resulted in a higher rate of molecular evolution at the isozyme level in the tetraploids.     

Restriction enzyme analysis of the chloroplast and nuclear 45s ribosomal DNA ofAllium sectionsCepa andPhyllodolon (Alliaceae)

Estimates of the phylogenetic relationships among cultivated and wildAllium species would benefit from identification of molecular characters. Restriction enzyme analysis of the chloroplast DNA (cpDNA) of the bulb onion (Allium cepa), Japanese bunching onion (A. fistulosum), wildAllium species in sect.Cepa andPhyllodolon, and the outgroupsA. ampeloprasum andA. tuberosum detected 39 polymorphisms.Allium cepa andA. vavilovii were identical for all characters. Cladistic analysis generated three most-parsimoniousWagner trees of 44 steps differing only in a zero-length branch.Allium fistulosum andA. altaicum (sect.Phyllodolon) comprised a monophyletic lineage separated from theA. cepa andA. vavilovii of sect.Cepa. The unresolved node was composed ofA. galanthum, A. roylei, and the lineage containingA. cepa, A. vavilovii, A. fistulosum, andA. altaicum. The clade containingA. altaicum, A. cepa, A. fistulosum, A. galanthum, A. roylei, andA. vavilovii remained resolved for strict consensus ofWagner trees of 48 steps or less.Allium pskemense andA. oschaninii were increasingly distant.Allium oschaninii has been proposed as the progenitor of the bulb onion, but was more closely related to the common progenitor of all species in sect.Cepa andPhyllodolon. Phylogenies estimated from cpDNA characters usingDollo parsimony resulted in a single most-parsimonious tree of 46 steps and agreed with phylogenies based onWagner parsimony. Polymorphic restriction enzyme sites in the 45s ribosomal DNA were not used to estimate phylogenies because of uncertain homologies, but are useful for identifying interspecific hybrids. The maternal phylogenies estimated in this study help to distinguish wildAllium species closely related to the bulb onion. Although not in agreement with classifications based on morphology, the phylogenies closely reflected crossability among species in sect.Cepa andPhyllodolon.     

Relationships between species ofLeymus,Psathyrostachys, andHordeum (Poaceae,Triticeae) inferred from Southern hybridization of genomic and cloned DNA probes

We have used total genomic DNA as a probe to size-fractionated restriction enzyme digests of genomic DNA from a range ofTriticeae species from the generaLeymus Hochst.,Psathyrostachys Nevski, andHordeum L., and hybrids betweenHordeum andLeymus to investigate their taxonomic relationships. Genomic Southern hybridization was found to be an effective and simple way to assess the distribution and diversity of essentially species-specific and common, repetitive DNA sequences, and is hence especially useful in evolutionary studies. The DNA sequences ofH. vulgare seem to diverge substantially from those ofH. brachyantherum, H. lechleri, H. procerum, andH. depressum. The genome ofThinopyron bessarabicum shows little homology to those of theLeymus species investigated, confirming thatT. bessarabicum is not an ancestral genome inLeymus. Although the genomes ofLeymus andPsathyrostachys share substantial proportions of DNA sequences, they include divergent repeated sequences as well. Hybridization with a ribosomal DNA probe (pTa 71) showed that the coding regions containing structural genes encoding the 18 S, 5.8 S, and 26 S ribosomal RNA were conserved among the species investigated, whereas the intergenic spacer region was more variable, presenting different sizes of restriction fragments and enabling a classification of the species. The rye heterochromatin probe pSc 119.2 hybridized to DNA fromH. lechleri andT. bessarabicum, but not to DNA from the other species investigated.     

Genetic relationships between European and Siberian larch,Larix spp. (Pinaceae), studied by allozymes. Is the Polish larch a hybrid between these two species?

Thirteen populations ofLarix decidua subsp.decidua and subsp.polonica, and three populations ofL. sibirica were investigated by starch-gel electrophoresis. In the populations assayed 61 alleles at 17 loci were revealed. The allozyme data support the earlier observations about close relationships between these two larch species. Nei's genetic distances betweenL. decidua andL. sibirica were relatively small (D = 0.057), however, almost five times larger, on average, than those between populations of the same species. Results obtained in this study disagree withBobrov's hypothesis about the hybrid origin of the Polish larch and suggest a direct origin from the European larch.     

Evolution and systematic relationships in theTriticeae (Poaceae)

The evolution and taxonomic relationships in theTriticeae are discussed with the view to highlight aspects of this agronomically important group of plants, which may be of interest to molecular biology. Some of these aspects are addressed in more detail in adjoining papers in which specific genomic loci have been examined at the DNA sequence or isozyme level. Aspects discussed include the systematics and geographic distribution of theTriticeae species, isozyme and chromosome pairing studies on some of the species as well as more recent developments in DNA analyses. A survey of the systematics of theTriticeae indicated that the genomic system ofLöve is probably the most useful starting point for interpreting molecular data even though the system has many problems from a taxonomic point of view. The geographical distribution ofTriticeae species, using both published and unpublished data, suggested that information of this type taken together with the theory of continental drift provides a broad time-span for considering data from DNA sequence studies. The significance, and modes of analyses, of isozyme studies were assessed because they often provide valuable characters in determining relationships between species. The main character underlyingLöve's andDewey's analyses of theTriticeae, namely chromosome pairing, is discussed with particular reference to isozyme studies to show that in some cases, such as species ofHordeum sensu lato, consistent relationships are obtained. Finally, new developments in understanding chromosome structure are considered in relation to the above variables in the taxonomy and evolution of theTriticeae.     

Isozyme studies in the genusCheirolophus (Asteraceae:Cardueae-Centaureinae) in the Iberian Peninsula, North Africa and the Canary Islands

The phylogenetic and phenetic analysis of 109 RAPD polymorphisms inS. nebrodensis, a perennial and self-incompatible endemic of four mountain ranges in Spain, andS. viscosus, a self-compatible annual widespread in Europe, as well asS. lividus, S. sylvaticus andS. vulgaris revealed a sister group relationship between the first two species. This result contrasts sharply with the earlier hypothesis based on isozyme variation thatS. viscosus originated from within a paraphyleticS. nebrodensis and that the two species represent a progenitor-derivative pair. After considering possible reasons for the sister group relationship found, including the possibility of rooting artefacts, it is concluded that neither the RAPD data nor the isozyme data allow to draw safe conclusions about the mode of speciation and therefore the relative age of the two species. As a consequence, the limited genetic variation ofS. viscosus in comparison toS. nebrodensis as revealed by both the RAPD and the isozyme data may reflect its population history, geographical distribution, reproductive ecology, or mode of dispersal just as well as its recent origin from a paraphyleticS. nebrodensis. The result of this study calls for a critical reexamination of other taxon pairs postulated to have a progenitor-derivative relationship on the basis of isozyme evidence.     

The 5S DNA units of bread wheat (Triticum aestivum)

A collection of 44 cloned 5S DNA units fromTriticum aestivum cv. Chinese Spring were grouped into 12 sequence-types based on sequence similarity and the respective consensus sequences were then produced. The relationship between these 12 consensus sequences (T. aestivum S 1-S 8 andT. aestivum L 1-L 4), together with two clones sequenced byGerlach andDyer, and the 5S DNA consensus sequences from diploidTriticum spp. were then determined by numerical methods. Both phenetic and cladistic analyses were carried out. The following wheat 5S DNA sequences were found to group with respective sequences from diploidTriticum spp.:T. aestivum S 4, S 6 withT. tauschii S;T. aestivum S 3 withT. monococcum S andT. monococcum S-Rus 7;T. aestivum L 1 andT. aestivum L-G&D withT. speltoides L;T. aestivum L 2, L 3 withT. tauschii L;T. aestivum L 4 withT. monococcum L andT. monococcum L-Rus 12. The analyses suggested that 5 out of the 65S Dna loci present in wheat were identified at the sequence level. The locus that could not be identified in this analysis was the5S Dna-B 1 locus. A group ofT. aestivum sequences (T. aestivum S 1, S 7, S 8, S-G&D) were found to be distinct from the other 5S DNA sequences in the data base. The existence of the distinct group of 5S DNA sequences suggests that there is a gap in our current understanding of wheat evolution with respect to the5S Dna loci.     

Relationships betweenOryza species (Poaceae) based on 5S DNA sequences

Relationships between 9Oryza species, covering 6 different genomes, have been studied using hybridization and nucleotide sequence information from the5S Dna locus. Four to five units of the major size class of 5S DNA in each species, 55 units in all, were cloned and sequenced. Both hybridization and sequence data confirmed the basic differences between the A and B, C, D genome species suggested by morphological and cytological data. The 5S DNA units of the A genome species were very similar, as were the ones from the B, C, and D genome-containing species. The 5S DNA ofO. australiensis (E genome) grouped with the B, C, D cluster, while the units ofO. brachyantha (F genome) were quite different and grouped away from all other species. 5S DNA units fromO. minuta, O. latifolia, O. australiensis, andO. brachyantha hybridized strongly, and preferentially, to the genomic DNA from which the units were isolated and hence could be useful as species/genome specific probes. The 5S DNA units fromO. sativa, O. nivara, andO. rufipogon provided A genome-specific probes as they hybridized preferentially to A genome DNA. The units fromO. punctata andO. officinalis displayed weaker preferential hybridization toO. punctata DNA, possibly reflecting their shared genome (C genome).     

Morphological and cytological evidence for polyphyletic allopolyploidy inArctostaphylos mewukka (Ericaceae)

The central Sierran tetraploidArctostaphylos mewukka Merriam has been reported to be an allopolyploid originating from the diploid species,A. patula Greene and eitherA. viscida subsp.viscida Parry orA. viscida subsp.mariposa (Dudley)P. V. Wells, although without conclusive evidence. Morphometrics and the verification and determination of chromosome numbers were used to substantiate the evolutionary relationships among these species. A closely related species,Arctostaphylos truei Knight, was also examined using these methods to determine its separability fromA. mewukka. The morphometric analyses support a hypothesis for a polyphyletic origin ofA. mewukka from different races ofA. viscida andA. patula. The chromosomal data, although inclusive, also support this hypothesis. The data do not support the recognition ofA. truei as a taxonomic entity separate fromA. mewukka at the species level.