Meiotic chromosome configurations in oocytes of <Emphasis Type="Italic">Cobitis taenia</Emphasis> and its polyploid hybrids

Diploid and triploid hybrid females of Cobitis as a rule produce unreduced eggs which mainly develop gynogenetically, but some of the eggs incorporate sperm genome and develop into triploids and tetraploids, respectively. Here, we observed for the first time the meiotic chromosomes in the germinal vesicles (GVs) of mature oocytes of three diploid C. taenia (2n = 48) and 20 allopolyploid females of Cobitis (18 triploid 3n = 74 and 2 tetraploid 4n = 99). The majority of GVs in diploid, triploid and tetraploid females contained 24, 74 and 96 or 99 bivalents, respectively. These results directly indicated premeiotic endomitosis as a mechanism underlying the formation of unreduced eggs in allopolyploid females of Cobitis.     

Ploidy-dependent survival of progeny arising from crosses between natural allotriploid Cobitis females and diploid C. taenia males (Pisces,Cobitidae)

Crosses between 21 triploid hybrid Cobitis females and 19 C. taenia (2n = 48) males led to viable progeny; whereas no embryonic development was observed in crosses with tetraploid males (4n = 98). The ploidy status of 491 progenies randomly selected with flow cytometry (316) or chromosome analysis (175) revealed an average of 55.2 % triploids and 44.8 % tetraploids, but the ratio of 3n versus 4n fish did change during development. In the first 2 days after hatching, approximately 65.1 % of tetraploid larvae were observed. Their number decreased significantly to 30.8 and 6.2 % on average during 2–5 and 10–15 months of life, respectively. The karyotype of tetraploid progeny (4n = 98) included 3n = 74 chromosomes of the parental female and n = 24 of C. taenia male. The number of tetraploid progeny indicated indirectly that about 66 % of eggs from 3n females were fertilized with C. taenia. The rest of the eggs developed clonally via gynogenesis or hemiclonally via hybridogenesis into triploids of the same karyotype structure as parental females. We have documented for the first time that (at least under experimental conditions) tetraploids are commonly formed, but are less viable than triploids, and a ratio similar to what is found under natural conditions is finally attained. The current explanation concerning the ploidy and karyotype structure of the progeny confirms that the eggs of 3n Cobitis females are not only capable of maintaining all chromosomes but are also capable of incorporating the sperm genome, thus creating the potential to produce tetraploids.     

Tetraploid induction of Mitracarpus hirtus L. by colchicine and its characterization including antibacterial activity

Tetraploid plants were successfully induced for the first time in Mitracarpus hirtus L., by overnight immersion of shoot meristems in 0.1 % colchicine solution, followed by in vitro culture leading to plant regeneration. Examination of ploidy level by flow cytometric analysis and counting chromosome number at metaphase confirmed that original diploid plant (WT1) contained chromosome number as 2n = 2x = 28, whereas 2n = 4x = 56 was observed in the tetraploids induced with colchicine treatment (CC102 and CC110). Thicker root formation, larger stomata (1.3–2 times), and lower density of stomata (1.7–4 times) were observed in these tetraploid plants. After transplantation to the pot, tetraploid plant (CC110) showed higher fresh weights of aerial part and leaves (1.5 and 1.4 times respectively) than diploid. However, the methanolic extracts from leaves of tetraploid line CC102 showed inhibition against human pathogenic bacterium, S. aureus while WT1 and CC110 showed no activity. GC–MS revealed 40 unique compounds present in CC102, but absent in WT1 and CC110. Through hierarchical clustering analysis the 40 unique compounds in CC102 formed a cluster group found to correlate with anti-S. aurens activity. These results suggested that tetraploid M. hirtus CC102 created in this study provides a novel source of compounds useful in fighting infectious disease.     

The role of triploids in the origin and evolution of polyploids of <Emphasis Type="Italic">Turnera sidoides</Emphasis> complex (Passifloraceae,Turneroideae)

Triploids can play an important role in polyploid evolution. However, their frequent sterility is an obstacle for the origin and establishment of neotetraploids. Here we analyzed the microsporogenesis of triploids (x?=?7) and the crossability among cytotypes of Turnera sidoides, aiming to test the impact of triploids on the origin and demographic establishment of tetraploids in natural populations. Triploids of T. sidoides exhibit irregular meiotic behavior. The high frequency of monovalents and of trivalents with non-convergent orientations results in unbalanced and/or non-viable male gametes. In spite of abnormalities in chromosome pairing and unbalanced chromosome segregation, triploids are not completely sterile and yielded up to 67% of viable pollen. Triploids that originated by the fusion of 2n?×?n gametes of the same taxon showed more regular meiotic behavior and higher fertility than triploids from the contact zone of diploids and tetraploids or triploids of hybrid origin. The reproductive isolation of T. sidoides cytotypes of different ploidy level is not strict and the ‘triploid block’ may be overcome occasionally. Triploids of T. sidoides produce diploid and triploid progeny suggesting that new generations of polyploids could originate from crosses between triploids or from backcrosses with diploids. The capability of T. sidoides to multiply asexually by rhizomes, would enhance the likelihood that a low frequency of neopolyploids can be originated and maintained in natural populations of T. sidoides.     

Cytology and embryology of the pompom weed,Campuloclinium macrocephalum (Eupatorieae,Asteraceae)

Campuloclinium macrocephalum (Less.) DC. is a perennial herb widely distributed in the New World, but introduced to South Africa, where it is commonly called “pompom weed”. This species is considered one of the most important weeds from Brazil and it has been included among the plant invaders of South Africa. Results of the meiotic and embryological analyses of six populations of C. macrocephalum are reported in this paper. The microsporogenesis analysis revealed five triploid (2n = 3x = 30) and one diploid (2n = 2x = 20) populations with a basic chromosome number x = 10. The diploid specimens showed regular meiotic behavior, but the triploid plants presented irregular chromosome pairing which result in the formation of univalents, bivalents and trivalents at diakinesis. In consequence, laggard chromosomes, unbalanced nuclei and micronuclei were observed in subsequent phases of meiosis. The embryological analysis showed that the triploid specimens of C. macrocephalum have embryo sac development from a nucellar cell (apospory), which indicates that these specimens are apomictic. Almost all cases of apomixis found in tribe Eupatorieae are diplosporous apomixis. Campuloclinium macrocephalum constitutes the second species of the tribe and the first of the genus with apospory as reproductive system. The aposporous apomixis combined with the presence of xylopodium would be two important factors responsible for the invasiveness of C. macrocephalum.     

Ecology of two cytotypes ofButomus umbellatus I. Karyology and breeding behaviour

Chromosome numbers were counted inButomus umbellatus from 99 localities in both Czech and Slovak Republics and the karyotype morphology was studied. Basic chromosome sets are asymmetrical and uniform among diploids (2n=26) and triploids (2n=39). Diploids occur less frequently than triploids in this region. Their clonal populations are usually fertile owing to self-compatibility. The clonal populations of triploids are selfincompatible and usually sterile. Thus, the different self-compatibility is the main biological character distinguishing diploids from triploids. Pollen of triploids is viable in spite of meiotic irregularities in pollen mother cells (PMCs). Hybridization both between cytotypes and between the different triploid genets may take place, if they occur sympatrically. Offspring having diverse chromosome numbers between diploid and triploid level can originate as the consequence of such hybridization.     

Possible pathways of the gene flow inTaraxacum sect.Ruderalia

Reproductive behaviour and the pathways of gene flow among ploidy levels were studied experimentally inTaraxacum sect.Ruderalia. Diploid, triploid and tetraploid individuals were sampled from mixed diploid — polyploid natural populations. 136 experimental hybridizations between the plants of different ploidy levels were performed. Seeds resulting from these crosses, those obtained from isolated anthodia as well as from open pollinated anthodia (both from cultivated and wild plants) were subjected to the flow-cytometric seed screening (FCSS) to determine ploidy levels in the progeny and to infer breeding behaviour of maternal plants. Three possible pathways of the gene flow were studied: (A) fertilization of sexuals by pollen of apomicts, (B) B_III hybrid formation, (C) facultative apomixis. Diploid maternal plants when experimentally crossed with triploid pollen donors produced diploids and polyploid progeny, while when pollinated with a mixture of the pollen of diploids and triploids or insect pollinated, no polyploids were discovered. It seems that in the mixture with the pollen of diploids, the pollen of triploids is ineffective. Tetraploids produce hybrids much easier with diploid mothers and their role in wild populations requires further study. Triploid mothers, even those with subregular pollen did not show traces of facultative apomixis. B_III hybrids were present in the progeny of both triploids and tetraploids, in tetraploids in quite high percentages (up to 50% of the progeny in some crosses).     

Interspecific hybridization in the genus Hieracium s. str.: evidence for bidirectional gene flow and spontaneous allopolyploidization

Although reticulation has indisputably played an important role in the evolutionary history of the genus Hieracium s. str. (Asteraceae), convincingly documented cases of recent interspecific hybridization are very rare. Here we report combined evidence on recent hybridization between two diploid species, Hieracium alpinum and H. transsilvanicum. The hybrid origin of the plants from the Romanian Eastern Carpathians was supported by additive patterns of nuclear ribosomal DNA polymorphism (ITS), an intermediate position of hybrid plants in principal coordinate analysis based on amplified fragment length polymorphism phenotypes (AFLP), and additivity at one allozyme locus. Flow cytometric analyses and chromosome counting showed that two hybrids were diploid (2n ~ 2x ~ 18) while one was surprisingly tetraploid (2n = 4x = 36). To our knowledge, this is the first record of spontaneous polyploidization following interspecific crossing in the genus. Allozyme data, especially the presence of unbalanced heterozygosity at one locus, suggest the origin of this tetraploid via a triploid bridge with subsequent backcrossing to H. alpinum. According to PCR-RFLP analyses of the trnT-trnL intergenic spacer, all H. ×krasani hybrids examined had the H. alpinum haplotype while H. transsilvanicum served as a pollen donor. The hybrids occurred at the locality with abundant H. alpinum plants where paternal H. transsilvanicum was missing. Previously reported instances of interspecific hybridization between the same parental taxa showed an opposite direction of crossing and relative abundance of parental taxa. This suggests that the direction of hybridization might be influenced by the frequency of parental taxa at the locality.     

Evidence for autoploid evolution in the artemisia ludoviciana complex of the Pacific Northwest

TheArtemisia ludoviciana complex of the Northwest is considered to be an intervarietal autoploid complex on the basis of evidence obtained from cytogenetic analysis. The evidence includes the occurrence of chromosomal races within all but two of the inclusive taxa, the degree and constancy of multivalent formation in the polyploid races, and the high degree of homology among the genomes of the various taxa as demonstrated by the pairing relationships in the F₁ progeny. Both triploid and tetraploid progeny were produced in diploid-tetraploid crosses, and the tetraploid offspring were fully as fertile as the natural tetraploids. The triploids, on the other hand, produced very few viable pollen grains. The production of tetraploid offspring in interracial crosses could provide a mechanism for gene flow from the diploid to the tetraploid population. With the observation of both diploid and tetraploid populations ofA. douglasiana, in addition to the well-known hexaploid, a reasonable doubt is cast upon the putative amphidiploid origin of the hexaploid via hybridization betweenA. suksdorfii andA. ludoviciana.     

Impaired male meiosis, morphology and distribution pattern of different cytotypes of Bupleurum lanceolatum Wall. (Apiaceae) from the Western Himalayas

Detailed male meiosis, critical morphological observations and distribution pattern of diploid as well as tetraploid cytotypes of the Western Himalayan species, Bupleurum lanceolatum have been evaluated at present. A diploid (n = 8) cytotype is reported from Kashmir, whereas, both diploid (n = 8) and tetraploid (n = 16) cytotypes are available from two districts Kangra and Sirmaur of Himachal Pradesh. Out of these, the tetraploid cytotype makes new addition for the species on a worldwide basis. As per behavior, a tetraploid cytotype is characterized by abnormal meiosis leading to high pollen sterility and size variation of the pollen grains. Morphologically, tetraploids are noted to be luxuriant in comparison to the diploids.     

Genome relationships in the diploid oats

Genome relationships between the three diploid oats, Avena strigosa (S.), A. longiglumis (L.) and A. prostrata (P.) were studied by chromosome pairing in diploid hybrids and in synthetic triploids and tetraploids combining these genomes. Fairly regular pairing in the diploid hybrid and typical autopolyploid behavior in the triploids and in the amphidiploid suggest small differentiation in the chromosome architecture of A. longiglumis and A. prostrata. A. strigosa diverges from the other two oats by complex chromosome rearrangements. Conspicuous preferential pairing took place in triploids with SSL, SSP and SPP genomic constitution. The low bivalent frequency in the SLL triploid suggests that preferential pairing in triploids with two S genomes is not a consequence of chromosome rearrangement but is rather of genetic origin. The presence of the three genomes in a triploid or a tetraploid caused considerable meiotic irregularities suggesting a better pairing competition of the S genome.     

二倍体鲫鲤F2产生不同倍性卵子的证据

在检测到鲫鲤F2产生3种不同大小(直径分别为0.13 cm,0.17cm和0.2 cm)类型的卵子基础上,进行了F2(♀)×红鲫(♂)及F2(♀)×四倍体鲫鲤(♂)的交配实验.通过染色体计数和流式细胞仪分析,在F2(♀)×红鲫(♂)后代中获得了四倍体、三倍体、二倍体鱼;在F2(♀)×四倍体鲫鲤(♂)后代中获得了四倍体和三倍体鱼.这两个交配组合后代中出现的不同倍性的鱼类为证明鲫鲤F2能产生三倍体、二倍体和单倍体卵子提供了进一步证据.F2(♀)×红鲫(♂)中雄性四倍体鱼的存在说明在四倍体后代中存在基因型为XXXY的个体.对上述两个交配组合后代的四倍体鱼和三倍体鱼的性腺结构观察表明四倍体鱼是可育的,而三倍体鱼是不育的.作者认为鲫鲤F2能够产生二倍体和三倍体卵子与核内复制机制和生殖细胞的融合有关.     

Morphometrical and acoustical comparison between diploid and tetraploid green toads

In the present paper we compare, on the basis of morphometrical characters and acoustical properties of the advertisement calls, a sample of 158 male green toads (Bufo viridis complex) collected in 12 breeding populations of south Kazakstan and north Kyrgyzstan. The samples of three populations resulted in only diploid toads (2n = 22), those of eight populations in only tetraploid toads (2k=44) whereas in one locality diploid, tetraploid and many triploid toads were collected. Diploid toads show significantly larger body size and proportionally larger head and shorter limbs than both tetraploids and triploids, whereas no evident morphometrical differences were observed between triploids and tetraploids. Diploid advertisement calls have spectral and temporal properties that significantly differ from those of both triploid and tetraploid advertisement calls. In particular, diploids produce significantly longer calls with higher pulse-rates and lower frequencies than those of tetraploids. We address the question of the factors that could be responsible for these differences and we discuss four hypotheses: (1) the direct effect of polyploid mutation, (2) genetic drift, (3) reproductive character displacement and (4) environmental selection.     

Mating interactions between coexisting dipoloid, triploid and tetraploid cytotypes ofHieracium Echioides (Asteraceae)

Experimental crosses between diploids, triploids and tetraploids ofHieracium echioides were made to examine mating interactions. Specifically, cytotype diversity in progeny from experimental crosses, intercytotype pollen competition as a reproductive barrier between diploids and tetraploids, and differences in seed set between intra- and intercytotype crosses were studied. Only diploids were found in progeny from 2x × 2x crosses. The other types of crosses yielded more than one cytotype in progeny, but one cytotype predominated in each cross type: diploids (92%) in 2x × 3x crosses, tetraploids (88%) in 3x × 2x crosses, triploids (96%) in 2x × 4x crosses, triploids (90%) in 4x × 2x crosses, tetraploids (60%) in 3x × 3x crosses, pentaploids (56%) in 3x × 4x crosses, triploids (80%) in 4x × 3x crosses and tetraploids (88%) in 4x × 4x crosses. No aneuploids have been detected among karyologically analyzed plants. Unreduced egg cell production was detected in triploids and tetraploids, but formation of unreduced pollen was recorded only in two cases in triploids. Triploid plants produced x, 2x and 3x gametes: in male gametes x (92%) gametes predominated whereas in female gametes 3x (88%) gametes predominated. Cytotype diversity in progeny from crosses where diploids and tetraploids were pollinated by mixture of pollen from diploid and tetraploid plants suggested intercytotype pollen competition to serve as a prezygotic reproductive barrier. No statistically significant difference in seed set obtained from intra- and intercytotype crosses between diploids and tetraploids was observed, suggesting the absence of postzygotic reproductive barriers among cytotypes.     

Fitness differences among diploids, tetraploids, and their triploid progeny in Chamerion angustifolium: mechanisms of inviability and implications for polyploid evolution

Abstract. Theoretical models indicate that the evolution of tetraploids in diploid populations will depend on both the relative fitness of the tetraploid and that of the diploid-tetraploid hybrids. Hybrids are believed to have lower fitness due to imbalances in either the ploidy (endosperm imbalance) or the ratio of maternal to paternal genomes in their endosperm (genomic imprinting). In this study we created diploids, tetraploids, and hybrid triploids of Chamerion angustifolium from crosses between field-collected diploid and tetraploid plants and evaluated them at six life stages in a greenhouse comparison. Diploid offspring (from 2 x × 2 x crosses) had significantly higher seed production and lower biomass than tetraploid offspring (from 4 x × 4 x crosses). Relative to the diploid, the cumulative fitness of tetraploids was 0.67. In general, triploids (from 2 x × 4 x , 4 x × 2 x crosses) had significantly lower seed production, lower pollen viability, and higher biomass than diploid individuals. Triploid offspring derived from diploid maternal parents had lower germination rates, but higher pollen production than those with tetraploid mothers. Relative to diploids, the cumulative fitness of 2 x × 4 x triploids and 4 x × 2 x triploids was 0.12 and 0.06, respectively, providing some support for effect of differing maternal:paternal ratios and endosperm development as a mechanism of hybrid inviability. Collectively, the data show that tetraploids exhibit an inherent fitness disadvantage, although the partial viability and fertility of triploids may help to reduce the barrier to tetraploid establishment in sympatric populations.     

The role of tetraploids in the sexual-asexual cycle in dandelions (Taraxacum)

Apomictic plants often produce pollen that can function in crosses with related sexuals. Moreover, facultative apomicts can produce some sexual offspring. In dandelions, Taraxacum, a sexual-asexual cycle between diploid sexuals and triploid apomicts, has been described, based on experimental crosses and population genetic studies. Little is known about the actual hybridization processes in nature. We therefore studied the sexual-asexual cycle in a mixed dandelion population in the Netherlands. In this population, the frequencies of sexual diploids and triploids were 0.31 and 0.68, respectively. In addition, less than 1% tetraploids were detected. Diploids were strict sexuals, triploids were obligate apomicts, but tetraploids were most often only partly apomictic, lacking certain elements of apomixis. Tetraploid seed fertility in the field was significantly lower than that of apomictic triploids. Field-pollinated sexual diploids produced on average less than 2% polyploid offspring, implying that the effect of hybridization in the 2x-3x cycle in Taraxacum will be low. Until now, 2x-3x crosses were assumed to be the main pathway of new formation of triploid apomicts in the sexual-asexual cycle in Taraxacum. However, tetraploid pollen donors produced 28 times more triploid offspring in experimental crosses with diploid sexuals than triploid pollen donors. Rare tetraploids may therefore act as an important bridge in the formation of new triploid apomicts.     

Aneuploidy and genetic variation in the Arabidopsis thaliana triploid response

Polyploidy, the inheritance of more than two genome copies per cell, has played a major role in the evolution of higher plants. Little is known about the transition from diploidy to polyploidy but in some species, triploids are thought to function as intermediates in this transition. In contrast, in other species triploidy is viewed as a block. We investigated the responses of Arabidopsis thaliana to triploidy. The role of genetic variability was tested by comparing triploids generated from crosses between Col-0, a diploid, and either a natural autotetraploid (Wa-1) or an induced tetraploid of Col-0. In this study, we demonstrate that triploids of A. thaliana are fertile, producing a swarm of different aneuploids. Propagation of the progeny of a triploid for a few generations resulted in diploid and tetraploid cohorts. This demonstrated that, in A. thaliana, triploids can readily form tetraploids and function as bridges between euploid types. Genetic analysis of recombinant inbred lines produced from a triploid identified a locus on chromosome I exhibiting allelic bias in the tetraploid lines but not in the diploid lines. Thus, genetic variation was subject to selection contingent on the final ploidy and possibly acting during the protracted aneuploid phase.     

Inter-embryo competition in the progeny of autotriploid Aloineae (Liliaceae)

In reciprocal crosses between diploid and triploid Aloineae the progeny are largely diploid or diploid plus one or two chromosomes, but in reciprocal crosses between triploids and tetraploids they are tetraploid or nearly so. Thus the triploids contribute circa haploid gametes to the progeny when crossed with diploids but circa diploid gametes when crossed with tetraploids. These results are compared with those of a number of earlier workers. It is concluded that the bias in the frequency of progeny types towards diploidy or tetraploidy, depending on the ploidy level of the plant which is crossed with the triploid, is caused by inter-embryo competition. Those embryos with an endosperm/embryo factor of 1.5, the value found in normal diploid/diploid crosses having triploid endosperms, are selected in preference to those with factors higher or lower than 1.5.Inter-gamete competition also occurs among the euploid and aneuploid gametes produced by the triploids. This is more pronounced on the male side, because the degree of survival of aneuploid pollen from the triploids into the next generation is much lower than that of aneuploid egg nuclei.Non-reduction in the triploids gives rise to occasional pentaploid progeny in crosses with tetraploids, but it is more probable that in diploid/triploid crosses tetraploid progeny are the products of non-reduction in the diploid.     

Spontaneous polyploids in marginal populations of Alopecurus bulbosus Gouan (Poaceae)

Natural populations of A bulbosus from three sites in southern England have been investigated and two were found to contain triploid and tetraploid plants in addition to normal diploids. Meiosis in the triploids was characterized by a high frequency of trivalent formation but, in the tetraploids, multivalents were formed at low frequencies and bivalents were the most common configurations. It is suggested that these are true autopolyploids that have probably arisen through somatic doubling since tetraploid cell lines were found in some anther loculi. The meiotic behaviour of the tetraploid plants is explained on the basis of bivalent-promoting mechanisms being present in the diploid progenitors.     

Persistence,dispersal and genetic evolution of recently formed <Emphasis Type="Italic">Spartina</Emphasis> homoploid hybrids and allopolyploids in Southern England

In Southampton Water, UK, the recent (c. 150 years ago) interspecific hybridisation between Spartina alterniflora (2n = 6x = 62; A-genome) and S. maritima (2n = 6x = 60; M-genome) gave rise to the homoploid hybrid (S. × townsendii, 2n = 6x = 62), and subsequently to the invasive allododecaploid species S. anglica (2n = 12x = 120–124) that has since spread worldwide. To address the question of dynamics of mixed ploidy populations involving these plants, we analysed several Spartina populations (fifty one individuals) in Southern England, UK, one of which was the presumed place of origin of the homoploid hybrid (Hythe). Using a combination of flow cytometry and ribosomal DNA (rDNA) genotyping we were able to identify the genomic composition and ploidy level of each individual analysed. The data show that the homoploid hybrid still dominates the population at Hythe (82 % of individuals collected in that locality) since its origin in the nineteenth century. We also identified S. × townsendii for the first time on Hayling Island (66 % individuals), indicating dispersal beyond its likely origin. The fertile allododecaploid S. anglica was mainly found in populations outside the initial hybridisation site, on Hayling Island and at Eling Marchwood. Quantification of the rDNA contributions from each parental genome showed that the ratios were mostly balanced in S. × townsendii. However, two (3 %) S. anglica individuals analysed have lost nearly all M-genome homeologs, indicating extensive repeat loss. Such variation indicates that despite the presumed single allopolyploid origin of S. anglica and genetic uniformity at other loci, it has undergone substantial changes at the rDNA loci following genome duplication.