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.     

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.     

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.     

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.     

Development of triploid hybrids in asparagus breeding employing a tetraploid landrace

The development of triploid cultivars from crosses between the tetraploid landrace ‘Morado de Huetor’ and diploid commercial cultivars was studied in this work. Fertility of both 4x × 2x and 2x × 4x crosses, as measured by fruit set and number of seeds per fruit, and germination rate of triploid seeds, were studied in order to assess the viability of producing commercial seeds. Seven triploid hybrid progenies were included in a progeny test together with thirteen experimental tetraploid hybrids and four cultivars (‘Grande’, ‘UC157-F2’, ‘Purple Passion’ and ‘Morado de Huetor’). Marketable spear production and mean diameter of spears were evaluated in both 2007 and 2008. Crosses 4x × 2x were more successful than 2x × 4x, and a wide variation from high to very low fertility within tetraploid females was found. Germination rate of triploid seeds was similar to diploid ones. After 2 years of evaluation experimental hybrid triploids were more productive than tetraploids with values similar to the highest yielding diploid cultivar (‘Grande’). Spear diameter of triploid hybrids was in general thicker than the tetraploid hybrids and significantly thinner than the standard cultivars ‘Grande’ and ‘Purple Passion’ (tertraploid). These results suggest that the development of new triploid cultivars (4x × 2x) employing a tetraploid landrace from a different genetic background of current diplods could be interesting in asparagus breeding, broadening the spectrum of cultivated asparagus cultivars.     

Unreduced pollen frequencies versus hybrid production in diploid-tetraploid Vaccinium crosses

Summary Reciprocal crosses were made between highbush Vaccinium corymbosum L. cultivars (4X) and high, medium and low frequency diplandroid pollen producing diploid clones of V. elliotti (Chapm.) and diploid pubescent forms of V. corymbosum. Pollination of 3,342 flowers produced 15 hybrids-11 tetraploids and 4 triploids. All V. elliottii hybrids were tetraploid while each successful diploid-tetraploid V. corymbosum cross produced at least one triploid. The ratio of tetraploid hybrids to 100 pollinations (H: 100P) was considered to be the best criterion for assessing crossability. Vaccinium elliottii produced a total of 8 4X hybrids while 2X V. corymbosum only produced 3. Vaccinium elliottii was more successful as a female parent and diploid V. corymbosum only produced hybrids as male parents. High frequency diplandroid clones showed a lower mean crossability as pollen parents (H: 100P=0.2) than medium frequency diplandroid clones (H: 100P=0.8). As predicted, 0% diplandroid clones wer unsuccessful as pollen parents. Unreduced pollen production was essential for tetraploid hybrid production in 4X×2X crosses. There appeared to be no consistent, inherent pattern between 2n pollen and 2n egg production in a clone nor was a tetraploid genotype effect on crossability consistently observed.Paper No. 10781 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC., 27695-7601     

Modes of inheritance of two apomixis components,diplospory and parthenogenesis,in Chinese chive (Allium ramosum) revealed by analysis of the segregating population generated by back-crossing between amphimictic and apomictic diploids

To investigate the mode of inheritance of apomixis in Chinese chive, the degrees of diplospory and parthenogenesis were evaluated in F₁ and BC₁ progenies derived from crosses between amphimictic and apomictic diploids (2n = 16, 2x). The F₁ population was generated by crossing three amphimictic diploids 94Mo13, 94Mo49 and 94Mo50 with an apomictic diploid KaD2 and comprised 110 diploids and 773 triploids. All the diploid F₁ plants examined were completely or highly eusporous and completely syngamic. All the triploid F₁ plants examined were highly diplosporous and highly parthenogenetic. KaD2 could not transmit its high level of apomixis via monoploid pollen grains. The BC₁ population, generated by crossing 94Mo49 with apomictic triploids found in the F₁ offspring, exhibited heteroploidy; it comprised haploid, diploid, triploid, tetraploid and various aneuploid individuals. In this generation, clear segregation was observed between diplospory and parthenogenesis. Analysis of the BC₁ population suggests that diplospory and parthenogenesis are each controlled by single dominant genes, D and P, respectively. However, all the BC₁ plants characterized as parthenogenetic were diplosporous. The absence of phenotypically eusporous parthenogenetic plants can be explained by assuming that the presence of diplospory gene is a prerequisite for the parthenogenesis gene expression in Chinese chive.     

A tetraploid hybrid plant from 4x × 2x crosses in Vitis and its origin

To study the origin of unreduced (2n) gametes in diploid Vitis cultivars, we surveyed the occurrence of tetraploid hybrid seedlings from 40 interploid crosses with five tetraploid and seven diploid cultivars. A total of 250 seedlings from the interploid crosses were established through embryo culture and by seed sowing. In 20 2x × 4x crosses, no tetraploid hybrid seedlings were derived from 8,902pollinations. In 20 4x × 2x crosses, two tetraploid hybrid seedlings were obtained from 8,057 pollinations. Investigation of isozyme genotypes of the two tetraploid seedlings using three variable enzyme systems indicated that one of the two seedlings resulted from the union of a diploid egg with a 2n male gamete and that failure of second meiotic division resulted in the formation of the 2n male gamete. The percentage of giant pollen grains in `Muscat Bailey A', a pollen parent of the tetraploid seedling, was relatively high(about 5.9%) and 10.9% of the giant pollen grains germinated on agar medium. These results suggested that 2n pollen of diploid cultivars are useful for breeding tetraploid grape. This revised version was published online in August 2006 with corrections to the Cover Date.     

Species hybrids in Delphinium

In the past many attempts have been made to cross the red and yellow flowering Delphinium species with cultivars of D. elatum. The poor results however are an indication that for this combination there is a cross barrier, the more so as both artificial and natural species hybrids in the genus Delphinium occur frequently. This is confirmed by the present investigations and the data obtained show that the barrier in the first instance is due to a difference in chromosome number between the diploid species and tetraploid cultivars. When the diploid parent plants have been made tetraploid by means of colchicine, a second barrier appears to exist for some crosses in one given direction.It has been shown that hybridization of red-flowering D. cardinale tetra and D. nudicaule tetra with D. elatum cv. offers few prospects for Delphinium breeding; however, the species hybrid D. nudicaule x cardinale tetra (as ) is more promising. In the latter case of these threefold species crosses fertile progenies have been obtained producing the desired combinations orange and red combined with large and semidouble flowers.Although nothing can be said as yet of the hybrids of D. elatum cv. and the yellow flowering D. zalil tetra, produced in 1960, the introduction of the yellow flower colour via crosses with D. zalil x cardinale tetra is not possible on account of sterility of the F₁.Apart from twofold species hybrids, attempts to obtain threefold and fourfold species crosses along mutually different lines have also been successful.     

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.     

Polygon of crossability between eleven species of Ipomoea: section Batatas (Convolvulaceae)

Summary Crossability within Ipomoea section Batatas is complex because of genetic, cytogenetic and physiological interactions. This complexity is responsible for the fact that knowledge about phylogenetic relationships within this section remains preliminary. Between 1988 and 1991, studies of planting, pollination and evaluation of progenies were carried out at the facilities of CIP in La Molina and San Ramon, Peru. Self-compatibility was found in nine diploid species. Self-incompatibility was found in diploid I. trifida, tetraploid I. tiliacea and both tetraploid and hexaploid I. batatas. Sexual compatibility is though to be related to a multiallelic sporophytic incompatibility system which is expressed in the stigmatic papillae. Altogether, 4,162 cross pollinations were made between 11 species and 76 interspecific combinations of 110 possible combinations in a diallel 11×11 design. From these 76 interspecific combinations only 38 survived, and in these crossability (= proportion between number of fruits harvested/pollinations made) ranged from 0.01 to 1.00 at La Molina. At San Ramon, 11 out of 17 cross combinations were successful, and in these crossability ranged from 0.01 to 0.71. Crossability appeared to be influenced by latitude which is related to flowering synchrony. The early death of seedlings is hypothesized to be related to a genomic imbalance between embryo and endosperm. Maternal effects were found in crosses between diploids and tetraploids which produced triploid and diploid offspring, and in crosses between hexaploids and diploids which gave rise to hexaploid offspring. This study shows that I. trifida and I. x leucantha may act as bridge species for gene flow from wild Ipomoea species to the genepool of the sweetpotato.     

Tracing of 2n egg occurrence in perennial ryegrass (Lolium perenne L.) using interploidy crosses

Several genotypes of Lolium perenne (2n = 2x = 14) were screenedfor the production of 2n eggs. In each cross a diploid genotype, used asfemale parent, was pollinated with a tetraploid genotype. All interploidycrosses were made in isolation cages. Flow cytometrical analysis of theprogeny of 154 diploid plants revealed 28 tetraploid descendants. Theseoffspring plants were harvested on 15 different diploid mother plants.Amplified Fragment Length Polymorphism (AFLP) markers were used tocheck the parenthood of these tetraploids. Two seedlings died, theremaining plants could, after AFLP fingerprinting, be identified as meiotictetraploids. The results obtained allowed to conclude that the tetraploidprogeny plants harvested on 15 diploid mother plants were the result ofthe fertilisation of an unreduced egg by a reduced pollen grain of thetetraploid plant.     

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.     

Germination in vitro of ryegrass pollen grains

Summary Pollen grains of diploid and tetraploid Italian, perennial and hybrid ryegrass (Lolium spp.) germinated on 20% sucrose solution with 10 ppm each of CaCl₂, H₃BO₃ and EDTA. Pollen tubes emerged within 2–3 minutes after sowing and completed full growth within 1 1/2 hours. The tube elongation showed the expected linear growth rate. The tubes of the haploid pollen grains grew faster (3.9 m/minute) than that of the diploid (3.3 m/minute). Diploid pollen grains germinated better on 25% than on 20% sucrose solution. Up to 25% of the diploid pollen produced two tubes.     

Hybrids of ryegrasses and meadow fescue and their value for grass breeding

1. 1.
   In attempts to intercross Lolium perenne and L. multiflorum with Festuca pratensis in a glasshouse, 13,284 emasculated flowers of diploid, triploid and tetraploid Lolium plants were provided with pollen of diploid and tetraploid Festuca plants. Only the combinations 2n\sx4n and 3n\sx4n produced hybrids, viz. on average 5.3 and 1.0 mature hybrid plants per 100 pollinated flowers. In crosses performed in winter these averages were 12.0 and 4.9 respectively.     
   
   

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).     

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.     

Towards breeding of triploid chamomile (Matricaria recutita L.) – Ploidy variation within German chamomile of various origins

German chamomile is an important medical plant with a long history of usage and a wide range of medical applications. Wild forms are diploid, whereas cultivated ones are diploid and tetraploid. Ploidy level variation within 15 origins (varieties, accessions, populations) of chamomile was investigated. Both naturally occurring triploids and those induced through directed crosses between diploid and tetraploid parents were identified and analysed, and these data could facilitate the exploitation of triploidy in chamomile as in other crop plants (fruit and ornamental plants).     

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.     

Trifluralin-mediated polyploidization of <Emphasis Type="BoldItalic">Rosa chinensis minima</Emphasis> (Sims) Voss seedlings

Many diploid rose species and cultivars possess valuable traits that can be introgressed into modern tetraploid cultivars. Interspecific, interploidy crosses are possible, but triploid hybrids typically have limited fertility, hindering further breeding and selection. Tetraploidizing diploids before mating with tetraploids can alleviate fertility barriers. The efficiency of trifluralin was investigated for polyploidization of Rosa chinensis minima (2n = 2x = 14) seedlings. Treatments were trifluralin at 0.086% and 0.0086%, colchicine (0.5%), and distilled water and contained 2% dimethyl sulfoxide and a surfactant. Approximately 5 l of the treatment solution was applied to the apical meristem of seedlings (N = 337, 82–85 per treatment) in the process of cotyledon expansion. Guard cell length, pollen diameter, and root tip squashes of rooted cuttings were used to detect polyploidy in meristematic layer (L)I, LII, and LIII, respectively. Trifluralin (0.086%) was the most effective treatment for polyploidization (LI 20.2%, LII 12.9%, LIII 12.9%), followed by trifluralin (0.0086%) (LI 10.6%, LII 7.1%, LIII 4.7%) and colchicine (LI 2.4%, LII 0%, LIII 0%). Polyploidization consistently occurred from LI inward. Polyploids as a group had reduced pollen stainability and a lower leaflet length to width ratio than diploids. In addition, two diploid seedlings were identified which produce 2n pollen. Considerations in selecting germplasm and generating somatically-induced polyploids from seedlings versus clones for use in breeding are discussed.