The origin of polyploids via 2n gametes in Vaccinium section Cyanococcus

Summary The production of 2n pollen (pollen with the sporophytic chromosome number) was evaluated in 4x and 6x taxa of Vaccinium section Cyanococcus. Mean frequencies of 2n pollen producers were 17.1% and 8.3% in natural 4x and 6x populations, respectively. The frequency of 2n pollen producers in the 4x species ranged from 8.6% (V. angustifolium) to 23.8% (V. pallidum). Level of 2n pollen production was genotypically variable (1% to 37.4%). The widespread occurrence of 2n pollen in 2x, 4x and 6x taxa suggests that sexual polyploidization was widespread and responsible for the origin of the polyploid species found in this genus. The frequency of 2n pollen producers was not significantly different between the 4x species and their putative 2x ancestors. These results support the origin of 4x and 6x taxa as a consequence of sexual polyploidization. Polyploids derived from sexual polyploidization would be expected to have increased fitness and flexibility due to the mode of 2n pollen formation. In blueberry species the predominant mode of 2n pollen formation is genetically equivalent to a first division restitution mechanism (FDR). FDR 2n pollen transmits a high percentage of the heterozygosity and a large fraction of the epistasis from the 2x parent to the 4x offspring.     

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.     

Genetic relationship between Echinochloa crus‐galli and Echinochloa oryzicola accessions inferred from internal transcribed spacer and chloroplast DNA sequences

Barnyardgrass, hexaploid Echinochloa crus‐galli, is considered to arise from the hybridization between tetraploid Echinochloa oryzicola and an unknown diploid species. The genetic relationship between E. crus‐galli and E. oryzicola was examined to investigate the position of E. oryzicola in the evolutionary process of E. crus‐galli, based on the nuclear DNA internal transcribed spacer (ITS) and the chloroplast cpDNA trnT‐L, trnL intron, and trnL‐F regions. New World E. crus‐galli was clearly separated from Eurasian E. crus‐galli and showed a close relationship to the American taxa, Echinochloa crus‐pavonis and Echinochloa walteri, in both the ITS and chloroplast DNA. The nrDNA ITS sequences indicated no differentiation between the Eurasian E. crus‐galli and E. oryzicola, in contrast to their clear divergence in the cpDNA sequence. The present results suggest that E. oryzicola is the male donor of E. crus‐galli.     

Relevance of sexual polyploidization for crop improvement – A review

Colchicine induced polyploids have not directly contributed for crop improvement in the past. On the other hand, the so-called natural polyploids, derived from the functioning of numerically unreduced(2n) gametes have been shown to be more relevant for crop improvement in many cases. Different types of cytological abnormalities during meiosis can give rise to 2n gametes and the genetic composition of these gametes is variable. Depending on the type meiotic abnormalities, various types of 2ngametes, such as first division restitution(FDR), second division restitution (SDR),indeterminate meiotic restitution (IMR) and post meiotic restitution (PMR) gametes,among others, have been described in recent years. For the improvement of autopolyploids such as potato, alfalfa,Vaccinium spp., and some of the fodder grasses, FDR gametes have been proved to be highly useful. However, the use of 2n gametes for the improvement of allopolyploid crops has received much less attention so far. Some of the investigations on allopolyploids, derived from Festuca-Lolium, Alstroemeria and Lilium species hybrids, have revealed that 2ngametes can be most useful for the introgression of alien genes and chromosomes into cultivars. An important feature of using sexual polyploidization in the case of allopolyploids is that introgression can be achieved through recombination due to genetic crossing-over between alien chromosomes as well as addition of alien chromosomes, which is extremely difficult or impossible to achieve in the case of colchicine induced allopolyploids. Because of the recent developments in the field of plant molecular biology, methods have become available for the analysis of 2ngametes and sexual polyploid progenies more accurately and to develop systematic breeding approaches. The methods include DNA in situ hybridization (GISH and FISH)and molecular mapping (AFLP, RFLP, RAPDs).In addition to providing basic information on the genetic and genome composition of the polyploid progenies, these methods can be potentially useful for a more efficient creation of desirable breeding material and cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Improved techniques for the induction and isolation of polyploids in the genus Fragaria

Summary The sensitivity of Fragaria seedlings to colchicine is dependent on the plant organ that is treated. Complete immersion of seedlings in a 1.5% colchicine solution results in total lethality, whereas the survival rates were more than 75% even at concentrations of 3.0% when only shoot apices were treated. High proportions of polyploids were isolated by treating shoot apices of seedlings with a 2.0% colchicine solution for 24–28 h. The dropper method is preferred to the tube method as it involves a minimum of manipulation and requires simple equipment. A differential response to colchicine was observed within and between different diploid species, diploid and tetraploid hybrids.     

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.     

Meiotic restitution mechanisms and 2n pollen formation in a Solanum tuberosum dihaploid and in dihaploid × wild species hybrids*

A Solatium tuberosum dihaploid that proved to be a 2n pollen producer, exhibited two types of meiotic abnormality involved in 2n pollen formation: premature cytokinesis at prophase II and parallel/tripolar spindles in meiosis II. For this reason such a dihaploid was preferentially used as a seed parent in crosses with the diploid species S. tarijense, S. sanctae-rosae, and S. phureja. Diploid interspecific hybrids were evaluated for male fertility, for 2n pollen production and stability over three years and for meiotic nuclear restitution mechanisms. Pollen stainability ranged from 6 to 85% and 2n pollen from 5 to 35% among hybrids. The meiotic mechanism generating 2n pollen was that of spindle abnormality in meiosis II and consequent dyad/triad formation. High correlation was found between fused/parallel spindles and dyads.     

Yield and tuber characteristics of 4x progeny from 2x×2x crosses

Summary Unrelated, unselected 2x haploid-species hybrids that produce 2n eggs, by second division restitution and 2n pollen due to first division restitution were intercrossed to generate 4x progeny. The 19, 4x families were compared with seven cultivars for tuber yield, tuber appearance and tuber set. The mean yield of the 4x families was 30% higher than the mean yield of the cultivars. The best five 4x families outyielded the best five cultivars by 57% and 69% at two locations. Cultivars were superior for tuber appearance and tuber set. The high yields of the 4x families are due to genetic diversity of the 2x parents and their ability to transmit that diversity to the 4x progeny utilizing 2n gametes. The potential of the 2x×2x breeding method is dependent on developing 2x hybrid clones that form 2n eggs and possess desirable tuber types; clones producing 2n pollen and good tuber quality are available.     

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.     

Initial acytokinesis during leaf protoplast culture of dihaploid and tetraploidSolanum tuberosum and diploidS. bulbocastanum

Summary Leaf protoplasts of dihaploid (2n=2x=24) and tetraploid (2n=4x=48)Solanum tuberosum, and diploidS. bulbocastanum (2n=2x=24) were cultured in liquid medium. The cultures were studied for early karyological changes during their development. Giemsa staining of spread preparations revealed extremely low percentages of protoplasts developing into calli with the parental chromosome number, and high percentages of acytokinetic cells. The nuclear divisions within a cell were synchronous which allowed the occurrence of spindle interaction, resulting in nuclear poly- and aneuploidization. Although polyploidization was also found in uninucleate cells, a major increase in the formation of true-to-type calli would certainly be established by the improvement of early cross wall formation.