Evaluation of FDR diploid and tetraploid parents in potato under two different day-length environments

Five tetraploid clones of potato were crossed to each of eight diploid first division restitution (FDR) 2n pollen producers and four tetraploid males using a line × tester mating design. A total of 59 families were obtained and evaluated under long days at Rhinelander (USA). A subset of 49 families were grown under short days at four Peruvian locations. Tuber yield (kg per plant) of 4x-2x families was similar to, or greater than, that of 4x-4x families in short day environments. The diploid clones had breeding values greater than or equal to those of the tetraploid clones. Only families derived from FDR 2n pollinators combined high tuber yield with stability and high dry matter content. Reciprocal recurrent selection would be the best breeding scheme, in which the diploids are testers of the tetraploids and vice-versa. Intra-population improvement could be achieved through phenotypic recurrent selection. The best materials from both breeding pools could then be crossed to produce tetraploid hybrids through unilateral sexual polyploidization (4x-2x crosses).     

A comparison of diploid and tetraploid turnips (Brassica campestris SSP. Rapifera)

Summary Five lines of tetraploid turnips and their diploid progenitors were examined. Diallel crosses were made at each level of ploidy and trial results showed that in all cases the diploids outyielded their respective tetraploids for dry matter production.     

Analysis of parental effects on mean vine maturity and chip colour of 4x.2x potato progenies

Summary The parental effects on progenies from potato 4x.2x(FDR) crosses were analyzed to determine anether and to what extent selection criteria used for selecting tetraploid breeding lines have to be adjusted when selecting diploid breeding lines. For vine maturity and chip colour multiple regression analysis of progeny means on means of diploid and tetraploid parents revealed high coefficients of determination (R²) for both characters, indicating good predicting power of the performance of both diploid and tetraploid parents on the performance of their 4x.2x progenies. For vine maturity the multiple regression slope for the tetraploid perents was significantly larger than for the diploid parents, and progeny means were towards the tetraploid parents. This indicates that selection criteria for vine maturity may be less stringent at the diploid level. However, diploids may be earlier maturing than tetraploids and therefore selection criteria can better be similar at both ploidy levels. For chip colour, multiple regression slopes for diploid and tetraploid parents were similar and progeny means were towards the darkest coloured parent irrespective of its ploidy level. This indicates that selection of breeding lines for chip colour can be conducted similarly at both ploidy levels.     

Aneuploids of perennial ryegrass (Lolium perenne)

Summary Aneuploid plants of perennial ryegrass (Lolium perenne L.) with 2n=15 to 30 chromosomes were obtained by crossing a near-triploid (2n=3x+1=22) with a diploid or on open-pollination with diploids and tetraploids. Aneuploids occurred with a frequency of 83% in near triploid × diploid progeny and 92% on open-pollination with diploid and tetraploid plants. Aneuploid plants with 15 to 18 chromosomes resembled diploids in morphology and those with 19 to 30 chromosomes were akin to tetraploids. Meiotic studies suggested that most aneuploid plants resulted from transmission of aneuploid egg cells (n=8 to 23). Aneuploid plants with 2n=27 to 30 chromosomes in the progeny of 22×14 cross originated from unreduced egg cells. Plants with 19 to 21 chromosomes were recovered only by immature seed culture. Aneuploid plants with 26 to 30 chromosomes and triploids (2n=21) had higher pollen fertility and bigger seeds than plants with 15 to 22 chromosomes.     

Cytogeography and variation of stomatal size of Paspalum Glaucescens (Gramineae; Paniceae) in Southern Brazil

Paspalum glaucescens belongs to the informal group Plicatula, reproductively characterized by the dominance of tetraploid apomitic lines in most of its populations, with rare diploid, sexual counterparts. The species shows high phenotypic variation. Twenty nine Southern Brazilian accessions were cytologically and morphologically analysed. Most of the accessions were tetraploid (2n = 4x = 40). Meiotic study of three tetraploids showed their irregular behaviour. Eight accessions presented the diploid level (2n = 2x = 20).This can be considered a very high frequency of diploids in a member of the Plicatula group. While the tetraploids are concentrated at the highest elevations, diploids were detected mostly in the lowlands. Average stomatal sizes were quite distinct on different ploidy levels, being larger in tetraploids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Polyploidy and sexual polyploidization in the genus Vaccinium

Genus Vaccinium,consisting of blueberries, cranberries, lingonberries and many related wild species, includes diploid, tetraploid, and hexaploid species. Most evidence indicates that the tetraploid species are autotetraploids, with non-preferential bivalent chromosome pairing. Although homoploid interspecific crosses usually produce numerous fertile hybrids if the parents are from the same section of the genus, inter sectional crosses at the diploid level normally produce no seedlings, weak seedlings, or seedlings that are very low in fertility. There is a strong but not complete triploid block within Vaccinium. Even with insections, tetraploid × diploid (and the reciprocal) crosses normally give only a few tetraploid and a few triploid hybrids. Hexaploid × diploid crosses within sections are very hard to make, and the few hybrids that have been obtained are pentaploid. The frequency of 2n gametes varies,both among genotypes within species and among species. Vaccinium pollen is normally shed in tetrads, and the frequency of large pollen grains shed in dyads can be used to estimate 2n gamete frequency. Cultivated blueberries occur at both the tetraploid and the hexaploid levels, and there are important genetic resources in the diploids. Unreduced gamete production has permitted transfer of genetic material from the diploid to the tetraploid level and from the diploids and tetraploids to the hexaploid level via triploid hybrids. Intersectional crosses can occasionally produce tetraploid Vaccinium hybrids that appear to behave as amphidiploids and have medium to high fertility. CommercialVaccinium cultivars are normally propagated by cuttings. Intersectional hybridization, chromosome doubling, and asexual propagation could permit the production of novel hybrid combinations with value as ornamentals or in fruit production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Bilateral polyploidization inDactylis glomerata L. subsp.lusitanica: occurrence,morphological and genetic characteristics of first polyploids

Summary InDactylis glomerata L. subsp.lusitanica, triploid and tetraploid plants were obtained by bilateral sexual polyploidization in crosses between diploid parents known to produce 2n gametes. The polyploid and diploid progeny were compared for allozyme diversity (allele number and heterozygosity), phenological (pollen fertility, inflorescence emergence date), cellular (stomatic cell size) and morphological characters (vegetative biomass, seed weight, total seed number per plant, panicle number and seed number per panicle length) in four contrasting growing conditions. Stomatal cell size and the total number of alleles were significantly lower in diploids than in polyploids which sometimes recovered more than two distinct alleles at a locus via 2n gametes possessing heterozygous loci. Pollen fertility was similar in diploids and tetraploids, but significantly lower in the triploids. Seed weight was significantly higher in polyploids, although the average number of seeds per panicle length and the total number of seeds was higher in the diploid sibs. No phenological differences occurred among ploidy levels whereas such differences were observed inDactylis in all the natural areas of sympatry between diploids and their related polyploids. Our results provide evidence for an instantaneous effect of polyploidization on certain traits and suggest that natural selection would act subsequently on several other traits to allow the establishment of new polyploids.     

Tetraploid Westerwolths ryegrass

Summary Germinated seeds of the annual Westerwolths ryegrass were treated with colchicine-solutions of 0.05 to 0.4% at a temperature of 29°C in order to obtain autotetraploids. The best results were obtained after treatment for 2 hours with 0.1 and 0.2% solutions or 4 hours with 0.05%. By these procedures about 8% of the treated seeds produced mixoploid or tetraploid mature plants. A preliminary progeny test was performed with seed of outstanding C₁-plants.In spaced plants and in drilled rows the dry matter production of these C₂-families was on average equally as high as that of the most productive original diploid variety. The proportion of leaves was higher in both the first and in the second cut, while drought resistance was considerably better. The tetraploids came later into ear, tillering ability was smaller and mildew attack was slightly heavier. In samples from drilled and from space-planted rows seed production per inflorescence was found to be higher in the tetraploids than in the diploids. The seed of the tetraploids was larger.From the data available it is concluded that chromosome doubling offers favourable possibilities for the improvement of Westerwolths ryegrass. Some suggestions for a breeding programme are given.     

Identification of tetraploid regenerants from cotyledons of diploid watermelon cultured in vitro

Summary Adventitious shoots were obtained from the diploid watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] cultivars Dixielee, Jubilee II, Mickylee, Minilee, and Royal Sweet by culturing excised cotyledons on shoot regeneration medium for six weeks. Tetraploid and diploid regenerants were identified by counting the number of chloroplasts per guard cell pair from leaves of regenerated plants. Cross fertilization of putative tetraploids with diploid pollinators and the production of triploid seed confirmed the efficacy of this approach. The mean number of chloroplasts for tetraploid regenerants was 19.1 whereas diploids averaged 11.2. These values were similar to tetraploid and diploid plants from seed. Ovary diameter, petal, and anther diameter of male flowers, and leaf length by width ratio were also good indicators of plant ploidy. Progeny obtained from self-fertile tetraploids of Mickylee were crossed with various diploid pollinators to produce triploid hybrid seed. All triploid plants from tissue culture-derived tetraploids produced fruit comparable in quality to fruit produced by currently-available triploid hybrids, demonstrating that in vitro tetraploid induction can be used to produce high quality tetraploid plants for use in triploid hybrid seed production.     

Artificially induced polyploidization in Humulus lupulus L. and its effect on morphological and chemical traits

Chemically induced polyploids were obtained by the colchicine treatment of shoot tips of Humulus lupulus L. ‘Sybilla’. Flow cytometry revealed that most of the treatments resulted in the production of tetraploids. The highest number of tetraploids was obtained when explants were immersed in 0.05% colchicine for 48 h. A field experiment was conducted to compare diploid and tetraploid plants and assess the effect of genome polyploidization on the morphological and chemical characteristics. Tetraploids showed significant differences in relation to diploids. They had thinner and shorter shoots. The influence of chromosome doubling was also reflected in the length, width and area of leaves. The length of female flowers in the tetraploids was significantly shorter than that observed in diploids. Tetraploids produced a diverse number of lupuline glands that were almost twice as large as those observed in diploids. The most distinct effect of genome polyploidization was a significant increase in the weight of cones and spindles. Contents of major chemical constituents of hop cones was little affected by ploidy level. Total essential oils were significantly lower than those in diploids. However there was a significant increase in the proportion of humulene, caryophyllene and farnesene, oils desired by the brewing industry.     

Comparative Genetics of Floral Morphology in Diploid and Allotetraploid Gossypium

The cultivated Gossypium A genome diploid species G.arboreum and G.herbaceum and the allotetraploid species G.hirsutum and G.barbadense share common morphology for various floral traits,which offers an ideal system in which to investigate genetic mechanisms that differentiate diploid and tetraploid genomes.For example,knowing how a single phenotype behaves in the diploids,and comparing the same trait with different dosage effects in the tetraploids,may provide a means to study inter- and intra-genomic interactions in the polyploid genome.     

Breeding of polyploid spinach

2x, 3x and 4x forms of spinach were compared in 1959, 1960 and 1961.At the same stage of development the tetraploids outyielded the diploids in total green matter and leaf blades in 1959 and 1961. In 1960 there was no significant difference. This year the tetraploids had a very small increase in green matter in the period between the two harvest dates. Yield of dry matter was determined in 1960 and 1961, and in both years the tetraploids outyielded the diploids. The rate of development decreased with increasing degree of ploidy. Compared at the same age, when the diploids were about to start bolting they were superior to the polyploids with regard to total yield, yield of leaf blades and yield of dry matter.The onset of flowering was considerably delayed in the polyploids. Generally the triploids were intermediate between the diploids and the tetraploids. The triploids and tetraploids had a larger proportion of monoecious plants, and a smaller proportion of males than the diploids.Report No. 27     

Evidence of sexuality in induced tetraploids of Brachiaria brizantha (Poaceae)

Difficulties in obtaining new breeding lines of Brachiaria (Trin.) Griseb., an important forage grass in Brazil, are mostly related to differences in ploidy among the accessions, and to apomixis, an asexual mode of reproduction. Usually, sexual accessions are diploid while apomicts are polyploid. Induced tetraploids of Brachiaria brizantha (A. Rich.) Stapf have been successfully obtained and this paper presents the results of a study of their reproductive modes and fertility. Despite frequent meiotic aberrations during microspore development, the induced tetraploids produced viable pollen and produced progeny after controlled self-pollination. Similarly to the original diploid sexual progenitor, embryo sacs of the Polygonum type with confirmed meiotic origin were present in the induced tetraploids suggesting chromosome doubling did not alter the reproductive mode. The embryo sac of the Polygonum type was also observed in progenies obtained after self and open pollination. Nevertheless, embryo sacs of the Polygonum and the Panicum types within the same ovule were observed in some progenies obtained after open pollination, probably having resulted from hybridization with tetraploid, apomictic plants. Indeed, the compatibility of the progeny with tetraploid, apomictic B. brizantha was confirmed by the formation of mature caryopses after controlled pollination. Evidence is presented that the induced tetraploids and their progeny are sexual plants and that they are compatible with natural tetraploid B. brizantha. The induced tetraploids will be useful for analyses of apomictic inheritance as well as in the development of sexual tetraploid lines in Brachiaria breeding programs.     

Isozyme variation between diploid and tetraploid cytotypes of Glycine tabacina (Labill.) Benth.

Summary Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids.     

Bilateral sexual polyploidization in potatoes

Summary Bilateral sexual tetraploidization was achieved by intercrossing diploid Phureja × haploid Tuberosum hybrids. Both parents contribute gametes with the somatic number of chromosomes (diplogynoids and diplandroids). More than 100 tetraploid progeny appeared in nine 2x–2x families. The tetraploids are generally more vigorous and significantly outyield their diploid full-sibs. The superiority of the tetraploids is interpreted on the basis of the mode of diplandroid (2n pollen) formation by first meiotic division restitution (FDR). Heterozygosis, epistasis and genetic diversity, if present in the parental diploid genotype, are largely maintained in the 2n gametes formed by FDR. and syngamy brings about more opportunities for heterotic responses in conjunction with polyploidization. Differences in tuber yield among tetraploid clones within families were found to be statistically significant (P <0.05). Thus, unlike somatic chromosome doubling, which can merely increase chromosome numbers, or somatic hybridization, which could in addition bring about heterosis, sexual polyploidization is also associated with genetic variability. The degree of inbreeding and the genotypic structure in the population of newly arisen tetraploids is discussed. The relationship among asexual reproduction, preservation of favorable nonadditive genetic effects by FDR. restoration of sexuality, and polyploidization is explored. The possible evolutionary significance of these factors is emphasized.     

Production of intraspecific aneuploids in the genus Solanum. 1. Doubling the chromosome number of seven diploid species

Summary The variation which persisted in interspecific aneuploids even after one or two generations of backcrossing prevented the morphological distinction of the aneuploids from their diploid sibs. An attempt was made to unmask the effects of aneuploidy by producing aneuploids within the more uniform genetic background of a single species.The procedure to produce trisomics and aneuploids of diploid species was to vegetatively double clones or seed of the species with colchicine, cross the induced tetraploids to diploids to produce triploids, and then cross the triploids to the original diploids. An attempt was made to use the same diploid clone and/or introduction of a species in all successive stages of the procedure in order to obtain as much uniformity as possible. This was largely unsuccessful because self-incompatibility and inbreeding reduced seed set. Seventy-three different tetraploids were obtained from seven species. Tuber treatments were noticeably unsuccessful in producing doubled plants. The two vegetative treatments were most successful and produced 41 different tetraploid clones while 30 doubled plants were obtained from four seed treatments.Morphologically the tetraploids resembled the diploid and there was no noticeable increase in size of leaflets and floral parts in the tetraploid. The average number of chloroplasts in ten pairs of guard cells was a distinctive feature which could efficiently be used in distinguishing the induced tetraploid plants from diploids. The tetraploids were generally less fertile than their diploid counterparts and the majority of them exhibited self-compatibility in comparison to self incompatibility of the diploids.Cooperative investigations of the Agricultural Research Service, U.S. Department of Agriculture and the Wisconsin Agricultural Experiment Station. Supported in part by grants from the Research Committee of the Graduate School, University of Wisconsin.     

Crossability relationships of andean potato varieties of three ploidy levels

Summary The formation of triploids in 4x×2x crosses was studied in primitive potato varieties from the Andes of Peru and Bolivia. Triploid frequencies were as high as 80–100% in some crosses, although the actual number of seeds was low. The triploid block varied according to the tetraploid parent, and was never entirely absent. The likelihood of triploid formation was studied in 4x×2x crosses where pollen from both tetraploids and diploids of known genotype was used. Even though the diploids were chosen for their non-production of 2n gametes, as detected in pollen grain size, 66% of the hybrids were tetraploid. The amount and probability of gene-flow between the tetraploid and diploid gene-pools, either directly through 2n gametes or through triploid bridges was examined. Most of the 20 cultivars of S. x chaucha produced seed in crosses with tetraploids and diploids, although the fertility of the latter crosses was four times higher. The female fertility of some of the triploids thus allows them to exchange genes with the tetraploids and diploids, although gene-flow occurs more rapidly through direct transfer from diploids to tetraploids via 2n gametes.     

Influence of flower and flowering characteristics on seed yield in diploid and tetraploid red clover

Tetraploid red clover (Trifolium pratense L.) often produces insufficient seed to be economically interesting. Numerous studies have identified poor pollination due to long corolla tubes as its cause, but other studies disagree. Therefore, we investigated seed yield, corolla tube length and flowering traits in 244 plants from five diploid and five tetraploid cultivars during 2 years. Tetraploids produced fewer seeds, fewer flower heads and fewer seeds per head, but an equal number of flowers per head when compared to diploids. Although corolla tubes were longer in tetraploid than in diploid red clover (P < 0.001), no correlation between corolla tube length and seed yield was detected. Therefore, the corolla tube length is likely neither the main nor only cause of low seed yield in tetraploid red clover. Instead, three putative causes for low seed yield were identified: tetraploids produced less inflorescences per plant, possessed a lower degree of determinacy and had markedly less seeds per head when compared to diploids. Possible causes for such differences include lower pollen viability and higher rates of embryo abortion.     

Vitamin C,flower color and ploidy variation of hybrids from a ploidy-unbalanced <Emphasis Type="Italic">Actinidia</Emphasis> interspecific cross and SSR characterization

Seedlings derived from an Actinidia interspecific cross between the hexaploid Actinidia chinensis var. deliciosa ‘Jinkui’ and the diploid male A. eriantha × A. chinensis var. chinensis ‘Chaohong’ hybrid were analyzed using flow cytometry, SSR markers and phenotypic observations. The results show that the leaf vitamin C content of this hybrid population has a mid-parent heterosis. Separation of flower color in the progeny was also observed, progeny with red flowers lighter than ‘Chaohong’, white flowers as in ‘Jinkui’ and intermediate types with a red base to the petals and white margins were all present. Flow cytometry analysis confirmed that most of the progeny were tetraploids, and molecular marker data showed that most of these tetraploid progeny had three alleles from the hexaploid parent and one allele from the diploid parent. UPGMA analysis based on the SSR markers showed that the diploid parent was completely separated from the hexaploid parent and all the progeny.     

Transmission of bacterial wilt resistance by First Division Restitution (FDR) 2n pollen via 4x×2x crosses in potatoes

Summary Resistance to bacterial wilt (Pseudomonas solanacearum) found in tuber-bearing Solanum species was transferred into a diploid potato breeding population. Simultaneous selections were made for agronomic characters, production of first division restitution (FDR) 2n pollen, and resistance. Diploid resistant genotypes were identified via inoculation with a virulent isolate (CIP-204) of race 3 of Pseudomonas solanacearum. These resistant diploid genotypes were crossed to susceptible tetraploid potatoes. An investigation was made to assess whether resistant diploid genotypes transmit resistance to bacterial wilt, which is a quantitatively inherited trait, to tetraploid potatoes via FDR 2n pollen. Tetraploid seedlings from 4x×2x crosses were inoculated with the same isolate CIP-204, and the percentage of surviving seedlings was scored. Some 4x×2x families from resistant diploid genotypes demonstrated a high level of survival rate. The transmission of bacterial wilt resistance was achieved by the use of FDR 2n pollen. It was speculated that a female x male interaction effect on the survival rate of the evaluated progeny may exist. Selecting proper 4x and 2x parents would be important for obtaining a higher frequency of transmission of resistance to bacterial wilt in the progeny.