SEGREGATION IN NEW ALLOPOLYPLOIDS OF GOSSYPIUM. IV. SEGREGATION IN NEW WORLD × ASIATIC AND NEW WORLD × WILD AMERICAN HEXAPLOIDS

Phillips , Lyle L. (North Carolina State Coll., Raleigh.) Segregation in new allopolyploids of Gossypium. IV. Segregation in New World × Asiatic and New World × wild American hexaploids. Amer. Jour. Bot. 49(1): 51–57. 1962.—The New World tetraploid cottons, G. hirsutum and G. barbadense, are natural amphidiploids (genome formula, 2 [AD]) combining species of the cultivated Asiatic (2A) and wild American (2D) groups of diploid cottons. Genetic segregation for marker alleles in New World × Asiatic and New World × wild American synthetic hexaploids have been determined. Average segregation for several loci in New World × Asiatic hexaploids is close to the autoploid 5:1 ratio, ranging from 5.1 to 6.8:1. Average segregation for 3 loci (L, Rd, and R₁) common to a series of New World × wild American hexaploids is: New World × G. raimondii, 9.3:1;–× G. harknessii, 16.4:1;–× G. armourianum, 17.4:1;–× G. aridum, 20.3:1;–× G. lobatum, 21.4:1–× G. thurberi, 32.9:1;–× G. gossypioides, 66.5:1. These data are discussed, and the method by which they were derived is compared with other cytogenetical means of discerning phylelic interrelationships among Gossypium species.     

A STUDY OF TRIPLOID AND 3 X – 1 ANEUPLOID PLANTS IN THE GENUS GOSSYPIUM

Cytological comparisons were made of triploid and 3x – 1 plants of Gossypium hirsutum (haplo 17 and 18) X G. aridum, G. armourianum, G. harknessii, and G. raimondii. Tests and observations led to these conclusions: (1) Chromosome conjugation varied significantly from plant to plant and date to date within plants. (2) The D genome chromosomes of G. hirsutum are closer in homology to G. raimondii than to the other species tested. (3) The chromosomes of G. aridum, have closer homology to the A genome of G. hirsutum than do the chromosomes of the other D species tested. (4) The D genome of G. hirsutum has a small translocation as compared to the genomes of the four D species studied.     

Production of fertile somatic hybrids of Gossypium hirsutum?+?G. bickii and G. hirsutum?+?G. stockii via protoplast fusion

Fertile somatic hybrids were obtained via symmetric electrofusion of protoplasts from two combinations of tetraploid cotton (G. hirsutum cv. Coker 201, AD genome) and diploid wild cottons G. bickii (G genome) and G. stockii (E genome), respectively. Observation by morphological, flow cytometric analysis, chromosome counting and RAPD analysis of the tested hybrids of Coker 201 + G. bickii and Coker 201 + G. stockii confirmed the regenerated plants as hybrid status. Cytological investigation of the metaphase root-tip cells revealed there were 78 chromosomes in the hybrids. Flow cytometric analysis showed the tested plants had a relative DNA contents close to the total DNA contents of the two parents. RAPD analysis revealed the hybrids contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the hybrids was intermediate between the two fusion partners. The hybrid plants were successfully transferred to the soil, and they bloomed and set bolls. It is sure that the new hexaploids developed by cell fusion would contribute to cotton breeding through backcrossing with the elite genotypes of G. hirsutum.     

Chromosome association and pollen fertility of parental and interspecific hybrids of Lycopersicon esculentum X L. hirsutum and L. pennellii

Cytological studies were carried out on two wild species (L. hirsutum and L. pennellii) and the cultivated species (L. esculentum) of tomato and their F1 hybrids. Both parents and hybrids show a diploid chromosome number of 2n=24. The meiotic behaviour of the cultivated species showed a high degree of chromosome homology resulting in a high level of chiasmata frequency per bivalent. In contrast, the two wild species showed a slight increase in uniyalent frequency and a decrease in bivalent formation and chiasmata frequency. The meiotic behaviour of the hybrids showed a high level of univalents and low levels of bivalents as well as trivalents. Highly significant decreases in chiasmata frequency and increases in meiotic abnormalities, especially in the L. esculentum X L. pennellii hybrid, also were detected. The high meiotic irregularity and low chiasmata frequency recorded in the second hybrid indicated the disharmony and difference between its parental genomes and also served to predict its sterility. With regard to degree of pairing recorded in the hybrids, there is a possibility that sterility in such cases may refer to genetic factors in addition to the previously mentioned reasons. Pollen fertility showed no great difference between L. esculentum and L. hirsutum and their F1 hybrid, but a significant decrease was recorded in the L. esculentum X L. pennellii hybrid, which was clearly associated with high meiotic irregularity, low chiasmata frequency and chromosome association.     

POLLEN MORPHOLOGY AND SYSTEMATIC RELATIONSHIPS AMONG NEW WORLD SPECIES IN TRIBE ANTIRRHINEAE (SCROPHULARIACEAE)

The morphology of mature pollen grains among 12 New World genera in tribe Antirrhineae has been examined with light and scanning electron microscopy. Pollen from 29 American species have mean polar diam that range from 17 to 26 μm, have a tectate structure with perforate, microreticulate, or reticulate surface pattern, and are subspheroidal to prolate, and trizonocolporate with fusiform or narrowly oblong colpi that are free or occasionally fused at the poles. Among the genera, pollen size is positively associated with style length but not haploid chromosome number. Three major morphological categories are recognized based on grain shape, exine structural pattern, and colpus shape. Two of these morphotypes each characterize individual genera (Mohavea, Linaria). The other morphological category includes ten New World genera and has been divided into three subcategories based on variation in lumina diam and intermittent occurrence of fused colpi. Only Antirrhinum and Pseudorontium have species or populations with pollen in more than one subcategory. Concordance of morphological, anatomical, chromosomal, and palynological data suggests that Linaria and Mohavea are best placed in separate subtribes from other native American genera. Compared to Old World taxa in tribe Antirrhineae, the most common pollen type (subspheroidal, microreticulate, apocolpate, fusiform colpi) may be the most primitive. Pollen grains with fused colpi (Antirrhinum p.p., Pseudorontium) and perforate (Mohavea) or reticulate (Holmgrenanthe) structural patterns probably represent derived types.     

Bidirectional cytoplasmic and nuclear introgression in the New World cottons Gossypium barbadense and G. Hirsutum (Malvaceae)

Patterns of interspecific cytoplasmic (plastid and mitochondrial) and nuclear introgression are typically asymmetrical: cytoplasmic gene flow or “capture” is frequently observed without evidence of nuclear introgression. In contrast, nuclear introgression without concomitant cytoplasmic introgression has rarely been demonstrated. Gossypium barbadense L. and G. hirsutum L. have large indigenous ranges in the New World semiarid tropics, with an extensive area of sympatry in the Caribbean and Central America. Numerous accessions of both species were surveyed for diagnostic cpDNA restriction sites. These data, in conjunction with previous information on nuclear markers, lead to several conclusions: 1) introgression between G. hirsutum and G. barbadense is bidirectional for both nuclear and cytoplasmic genes; 2) patterns of introgression between the two species are not symmetrical—in G. barbadense, introgression of G. hirsutum alleles is largely restricted to modem cultivars and is uncommon in areas of sympatry; in contrast, introgression of G. barbadense alleles into G. hirsutum is relatively common in areas of sympatry and is rare in modem cultivars; 3) nuclear introgression is geographically more widespread and more frequently detected than cytoplasmic introgression. Several mechanisms may underlie the differential patterns of cytoplasmic and nuclear gene flow observed, including differential fitness of infraspecific and interspecific cytonuclear combinations and selection against female function in interspecific backcrosses. Possible explanations for the observed asymmetrical patterns of introgression include differences in population sizes combined with phenological differences that bias interspecific pollen transfer.     

CHROMOSOME HOMOLOGY IN SOME INTERCONTINENTAL HYBRIDS IN HIBISCUS SECT. FURCARIA

Ten kinds of interspecific hybrids were obtained involving the following species: H. surattensis L. (2x, genome constitution BB), H. sudanensis Hochr. (2x, GG), and H. rostellatus Guill. and Perr. (4x, GGHH) from Africa; H. furcatus Roxb. non Willd. (8x) from India and Ceylon; H. furcellatus Lam. and H. bifurcatus Cav. (both 4x, PPQQ) from South America; and H. heterophyllus Vent. (6x) from Australia. Chromosome pairing in pollen mother cells (PMC's) at metaphase I in the 4x hybrids H. bifurcatus-rostellatus and H. furcellatus-rostellatus indicated that the parents have one genome in common (Q = G or H). Hibiscus furcatus was shown earlier to have a B genome; hybrids of H. surattensis-sudanensis F₁ X furcatus were hexaploid, having received an unreduced gamete from their hybrid parent, and had approximately 36 II, 36 I in PMC's. The genome formula of H. furcatus may therefore be designated BBGGWWZZ. The hybrid H. rostellatus-furcatus (BGGHWZ) confirmed that H. furcatus has a G genome in common with H. rostellatus; pairing of the other three genomes was inconsistent, as was that in H. rostellatus-heterophyllus. Some samples of the latter approached 36 II, 36 I, expected if H. heterophyllus were GGHHJJ; other samples had less pairing. Hibiscus furcatus-heterophyllus hybrids apparently arose from unreduced gametes of H. heterophyllus and originated as decaploids rather than heptaploids; chromosome number was unstable in PMC's. Nevertheless, multivalents, especially trivalents, were frequent enough to suggest that H. furcatus and H. heterophyllus share G genomes. On the other hand, an 8x H. bifurcatus-furcatus hybrid, which apparently arose from an unreduced gamete of H. bifurcatus, had a low multivalent frequency. Hybrids were obtained of H. heterophyllus X sudanensis and H. surattensis-sudanensis X heterophyllus, but the plants were weak and were not analyzed cytologically. We suggest that the New World, African, Indian, and Australian genomes which retain a considerable degree of homology (G or H or both) were distributed by land prior to separation of the southern continents by continental drift.     

Validation of a cotton-specific gene, Sad1, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic cottons

Genetically modified (GM) cotton lines have been approved for commercialization and widely cultivated in many countries, especially in China. As a step towards the development of reliable qualitative and quantitative PCR methods for detecting GM cottons, we report here the validation of the cotton (Gossypium hirsutum) endogenous reference control gene, Sad1, using conventional and real-time (RT)-PCR methods. Both methods were tested on 15 different G. hirsutum cultivars, and identical amplicons were obtained with all of them. No amplicons were observed when DNA samples from three species of genus Gossypium, Arabidopsis thaliana, maize, and soybean and others were used as amplified templates, demonstrating that these two systems are specific for the identification and quantification of G. hirsutum. The results of Southern blot analysis also showed that the Sad1 gene was two copies in these 15 different G. hirsutum cultivars. Furthermore, one multiplex RT-quantitative PCR employing this gene as an endogenous reference gene was designed to quantify the Cry1A(c) gene modified from Bacillus thuringiensis (Bt) in the insect-resistant cottons, such as Mon531 and GK19. The quantification detection limit of the Cry1A(c) and Sad1 genes was as low as 10 pg of genomic DNA. These results indicat that the Sad1 gene can be used as an endogenous reference gene for both qualitative and quantitative PCR detection of GM cottons.     

FLORAL PIGMENTATION STUDIES IN THE GENUS GOSSYPIUM II CHEMOTAXONOMIC ANALYSIS OF DIPLOID GOSSYPIUM SPECIES

The chromatographic pigment arrays of nine diploid species (G. arboreum, G. anomalum, G. herbaceum, G. stocksii, G. sturtii, G. thurberi, G. gossypioides, G. raimondii and G. klotzschianum) were studied. Among the Old World cottons, G. sturtii of Australia was very different from the species analyzed. The two species of the Herbacea section (G. herbaceum and G. arboreum) were found to have very similar pigment arrays. Both G. anomalum and G. stocksii were more like the Herbacea species than any other species in the genus, but both G. anomalum and G. stocksii had unique pigment characteristics. Although the evidence obtained so far from pigmentation patterns suggests that some pairs of species are closely related, the pigment arrays do not support the classification of the New World diploids into more than one section. From analysis of pigments of interspecific hybrids and their parents, it was found that with a hybrid and one parent species the pigment array of the other parent species could be predicted. Using this approach, the pigment arrays of three New World diploid species were predicted.     

Genetic diversity and geographic pattern in early South American cotton domestication

Amplified fragment length polymorphism fingerprinting was applied to survey the genetic diversity of primitive South American Gossypium barbadense cotton for establishing a possible link to its pre-Columbian expansion. New germplasm was collected along coastal Peru and over an Andean transect in areas where most of the archaeological evidence relating to cotton domestication has been recorded. Gene bank material of three diploid (G. raimondii, G. arboreum, and G. herbaceum) and four allotetraploid cotton species (G. hirsutum, G. mustelinum, G. tomentosum and additional G. barbadense) was added for inter- and intra-specific comparison. Eight primer combinations yielded 340 polymorphic bands among the 131 accessions. The obtained neighbor joining and unweighted pair-group method with arithmetic means are in full agreement with the known cytogenetics of the tetraploid cottons and their diploid genome donors. The four tetraploid species are clearly distinct based on taxonomic classification. The genetic diversity within G. barbadense reveals geographic patterns. The locally maintained cottons from coastal Peru display a distinct genetic diversity that mirrors their primitive agro-morphological traits. Accessions from the northernmost coast of Peru and from southwestern (SW) Ecuador cluster basal to the east-of-Andes accessions. The remaining accessions from Bolivia, Brazil, Columbia, Venezuela, and the Caribbean and Pacific islands cluster with the east-of-Andes accessions. Northwestern Peru/SW Ecuador (the area flanking the Guayaquil gulf) appears to be the center of the primitive domesticated G. barbadense cotton from where it spread over the Andes and expanded into its pre-Columbian range.This publication is dedicated to Prof. Dr. Drs.h.c. Gerhard Röbbelen on the occasion of his 75th birthday     

OLMECA,A NEW GENUS OF MEXICAN BAMBOOS WITH FLESHY FRUITS

Olmeca, a new bamboo genus of two species is described from southern Mexico. It is unique among New World bamboos in its production of fruits that are large and fleshy, in contrast to the smaller, dry type—caryopsis or achene—that characterizes the other genera. The fleshy fruit is found in few other bamboos, among them Dinochloa, Melocanna, and Ochlandra. These Asiatic genera are not all closely related to each other nor to the new American genus, but do share a similar type of habitat: the tropical lowland rain forest, where ample water is available throughout the year. Besides the rare type of fruit found in Olmeca, the rhizomes of this genus develop necks up to 8 m or more long, an uncommon feature among any world bamboo of the sympodial type. The solitary culms of Olmeca grow widely spaced from one another because of these elongated, intervening necks. One species. O. recta, is known only from the mountain range of Los Tuxtlas, in Veracruz, where it grows on acidic soils derived from basalt; the other, O. reflexa, occurs on calcareous soils and has a wider range that encompasses parts of Veracruz, Oaxaca, and Chiapas.     

An interspecific somatic hybrid between upland cotton (<Emphasis Type="Italic">G. hirsutum</Emphasis> L. cv. ZDM-3) and wild diploid cotton (<Emphasis Type="Italic">G. klotzschianum</Emphasis> A.)

Somatic hybrids were produced through protoplast electrofusion between the tetraploid cotton Gossypium hirsutum L. cv. ZDM-3 and the wild diploid cotton G. klotzschianum. Hybrid plants were generated from 3 out of 24 callus lines that were derived from fused protoplasts. Hybrid plants were initially identified as somatic hybrids by ploidy analysis: the plants from the 3 callus lines had chromosome numbers near to sum of the two parents (78 = 52 + 26). The plants from the 3 lines were subsequently confirmed as hybrids by cytological, molecular, histological and morphological analyses. The morphology of hybrids was distinct from that of the parents, with elongated stigmas and malformed anthers lacking microspores and pollen, leading to male sterility. It is expected that the male sterility resulted from the high number of univalent and irregular multivalent chromosome pairings per meiocyte.     

Genetic mapping of a cross between Gossypium hirsutum (cotton) and the Hawaiian endemic, Gossypium tomentosum

The existence of five tetraploid species that derive from a common polyploidization event about 1 million years ago makes Gossypium (cotton) an attractive genus in which to study polyploid evolution and offers opportunities for crop improvement through introgression. To date, only crosses (HB) between the cultivated tetraploid cottons Gossypium hirsutum and G. barbadense have been genetically mapped. Genetic analysis of a cross (HT) between G. hirsutum and the Hawaiian endemic G. tomentosum is reported here. Overall, chromosomal lengths are closely correlated between the HB and HT maps, although there is generally more recombination in HT, consistent with a closer relationship between the two species. Interspecific differences in local recombination rates are observed, perhaps involving a number of possible factors. Our data corroborate cytogenetic evidence that chromosome arm translocations have not played a role in the divergence of polyploid cottons. However, one terminal inversion on chromosome (chr.) 3 does appear to differentiate G. tomentosum from G. barbadense; a few other apparent differences in marker order fall near gaps in the HT map and/or lack the suppression of recombination expected of inversions, and thus remain uncertain. Genetic analysis of a discrete trait that is characteristic of G. tomentosum, nectarilessness, mapped not to the classically reported location on chr. 12 but to the homoeologous location on chr. 26. We propose some hypotheses for further study to explore this incongruity. Preliminary quantitative trait locus (QTL) analysis of this small population, albeit with a high probability of false negatives, suggests a different genetic control of leaf morphology in HT than in HB, which also warrants further investigation.     

KARYOTYPE ANALYSIS IN SOME GENERA OF COMPOSITAE. VIII. FURTHER STUDIES ON THE CHROMOSOMES OF ASTER

Huziwara , Y. (Kobe U., Mikage, Kobe, Japan.) Karyotype analysis in some genera of Compositae. VIII. Further studies on the chromosomes of Aster. Amer. Jour. Bot. 49 (2) : 116–119. Illus. 1962.—The karyotypes of 3 Asiatic and 3 American taxa of Aster are reported here for the first time. These are: A. ageratoides subsp. sugimotoi (Kitamura) Kitamura 2n = 36, A. ageratoides subsp. (Taxon AIII) 2n = 72, A. himalaicus C. B. Clarke 2n = 18, A. ericoides L. 2n = 32, A. meritus A. Nels. 2n = 27, A. umbellatus Mill. 2n = 18. Two American taxa, namely, A. meritus and A. umbellatus, are considered to be ancestral taxa which have retained primitive karyotypes similar to those of Asiatic species of Aster.     

THE COLONIZATION OF DROSOPHILA SUBOBSCURA IN CHILE. II. CLINES IN THE CHROMOSOMAL ARRANGEMENTS

Drosophila subobscura is a Palearctic species that was first detected in the New World in Puerto Montt (Chile) in February 1978. Since that time, it has spread over a broad area and increased in population density. The South American populations exhibit a high level of chromosomal polymorphism: 20 different arrangements exist, distributed among five chromosomes. Chromosomal arrangement heterozygosity varies from 0.55 to 0.61 in the nine populations examined. Incipient clines in the frequencies of the arrangements are appearing; these clines follow the same latitudinal direction as in the Old World. Wing length significantly decreases with latitude, as it does in Europe. The colonization of South America by D. subobscura appears to be a major natural experiment with outcomes that duplicate the distributional patterns—in chromosomal polymorphism and in wing length—observed in the Old World, thereby strongly supporting the adaptive significance of these patterns. The data show a very rapid effect of natural selection promoting genetic differentiation among natural populations.     

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.     

TAXONOMIC AND GEOGRAPHIC DISTRIBUTION OF INTERNAL GEITONOGAMY IN NEW WORLD CALLITRICHE (CALLITRICHACEAE)

Internal geitonogamy (IG) is a unique self-fertilization system that occurs in the genus Callitriche. In IG self-fertilization is effected by pollen tube growth through vegetative tissues from the staminate to pistillate flowers. IG occurs in seven of the 22 species of Callitriche in the New World (C. heterophylla, C. heteropoda, C. lechleri, C. nubigena, C. rimosa, C. trochlearis, C. verna). Callitriche verna occurs in both the New and Old Worlds and represents the only known instance of IG in the latter region. IG is correlated with the amphibious growth habit and polyploidy, although not all polyploid amphibious species of Callitriche have IG. Correlations between IG and geographic distribution are not apparent. There is a tendency for species that possess IG to occur at higher elevations than those that lack this system.     

EXPERIMENTAL HYBRIDIZATION OF XANTHIUM STRUMARIUM (COMPOSITAE) FROM ASIA AND AMERICA. II. SESQUITERPENE LACTONES OF F1 HYBRIDS

Effects of hybridization on sesquiterpene lactones in Xanthium strumarium were studied for clues to the relationship of Old and New World populations. In crosses between indigenous Asiatic plants in the “strumarium” morphological complex and various American complexes that produce xanthinin as a major sesquiterpene lactone, the F₁ hybrids contained xanthinin and the related compounds, xanthanol, xanthatin, and xanthinosin. In other crosses with various American complexes that produce xanthumin, the stereoisomer of xanthinin, the F₁ hybrids contained xanthinin and xanthumin as well as xanthinin-related compounds and their stereoisomers, xanthumanol, deacetoxylxanthumin, and tomentosin. The Asiatic plants of “strumarium” from Hong Kong involved in the crosses produce approximately equal percentages of xanthinin, xanthatin and xanthinosin, but those from India contain only xanthinosin. The putative introduction of the American morphological complex, “chinense,” contains xanthumin as the major component but shows chemical diversity that indicates genes derived from the indigenous “strumarium” complex.     

INTERSPECIFIC INCOMPATIBILITY IN GOSSYPIUM. I. STEM HISTOGENESIS OF G. HIRSUTUM X G. GOSSYPIOIDES

The hybrid of Gossypium hirsutum X G. gossypioides develops normally until 10–12 nodes have been produced at which time neoplasms form in the cortex, phloem, and pith of the lower portion of the shoot axis. Neoplasms are characterized by irregular whorls of unusually large cells the central portions of which contain apparently cytolysed cells and a reddish-brown substance that probably represents the products of cytolysis. Neoplastic invasion is followed by inactivation of the vascular cambium, disorganization of the phloem and cortex, and tylosing and plugging of many xylem vessels. Subsequent growth of the shoot, which continues at a very slow rate for up to several years, involves the proliferation of parenchymatous tissue which becomes secondarily vascularized.     

SYNTHETIC HYBRIDS OF NEW WORLD AND OLD WORLD AGROPYRONS II. AGROPYRON RIPARIUM X AGROPYRON REPENS

Five hybrids were obtained from 12 seeds formed in 26 emasculated florets of A. riparium pollinated by A. repens. The hybrid plants were morphologically intermediate between the parents for vegetative and spike characteristics, although they resembled A. repens more closely than A. riparium. The 28-chromosome A. riparium parent behaved cytologically as an allotetraploid and formed an average of 13.98 II and 0.04 I in 94 cells at metaphase I. An average of 20.27 II and 0.36 IV were observed at metaphase I in 55 cells of A. repens, which was designated as a segmental autoallohexaploid. The hybrids contained 35 chromosomes and averaged 6.75 I, 12.49 II, 1.05 III, 0.01 IV, and 0.01 V in 162 cells interpreted at metaphase I. Bivalent chromosome pairing in the hybrids was attributed to autosyndetic pairing of 2 A. repens genomes and allosyndetic pairing between 1 A. riparium genome and 1 A. repens genome. Multivalent chromosome associations were attributed to structural hybridity. A. repens and A. riparium apparently share a genome in common, and this genome is the one responsible for rhizomes in both species. A. riparium was given a genome formula of R₂R₂SS; whereas the A. repens genome formula was written as R₁R₁X₁X₁X₂X₂, and the hybrid genome formula was designated as R₁R₂X₁X₂S. The “S” genome of A. riparium was derived from A. spicatum, and the “R” genome is the genome shared by A. repens and A. riparium. The origin and distribution of the so-called “X” genomes of A. repens remain unknown. The hybrids produced from 3 to 10% stainable pollen; however, no seed was set on the hybrids during 2 years in the field.