2n gametes in potato species and their function in sexual polyploidization

Summary The extent and pattern of polyploidy in the tuber-bearing Solanums varies among the many taxanomic series that have been identified in this subsection of Solanum. While several series appear to be entirely diploid, others exhibit a range of ploidy levels from 2x to 6x, and some contain only polyploid species.In many diploid, triploid and tetraploid species 2n gametes (gametes or gametophytes with the sporophytic chromosome number) have been detected. Both 2n eggs and 2n pollen occur. 2n gametes provide the opportunity for unilateral and bilateral sexual polyploidization. The genetic determination and consequences of sexual polyploidization strongly suggest that 2n gametes have been the major instrument in the polyploid evolution of the tuber-bearing Solanums. Somatic doubling of species and interspecific hybrids appears to be of very limited importance.New evidence for the occurrence of 2n eggs and 2n pollen in many species is reported, and data from the literature are added to illustrate the widespread distribution of 2n gametes throughout the subsection. A very high correlation is found between polyploidy and 2n gametes, and its significance is discussed. Proof is presented for the occurrence of alleles governing 2n pollen production in the cultivated tetraploids, providing additional evidence for the hypothesis that 2n gametes have been involved in their origin.Multiple unilateral and bilateral sexual polyploidizations are proposed for the origin of the cultivated tetraploids: this accounts for the large variability encountered in this group, which closely resembles that of the related diploids. Similar evolutionary pathways are hypothesized for the other polyploid complexes in the subsection.A scheme is proposed in which participation of both 2n and n gametes link together all ploidy levels in the tuber-bearing Solanums, thus overcoming the ploidy barriers and providing for gene flow throughout the sympatric species of the subsection.Paper No. 2093 from the Laboratory of Genetics. Research supported by the College of Agricultural and Life Sciences, a gift from Frito-Lay, Inc., and grants from NIH (GM 15422) and The International Potato Center. Plant materials, unpublished chromosome counts and technical help of IR-1 are gratefully acknowledged.     

In tergeneric crosses with Fragaria and Potentilla. II. Crosses between the progeny of Fragaria moschata x Potentilla fruticosa and the original parents

Summary An aneuploid hybrid (2n=23) of Fragaria moschata (2n=42) and Potentilla fruticosa (2n=14) was backcrossed with pollen of both parents, separately and combined in a pollen mixture. Seven vigorous progeny were obtained. The origin of the exeptional chromosome numbers, 2n=44, 49, 63, 63, 65, 67, 67, is discussed, and it is shown that each of the numbers could be produced by the fertilisation of unreduced and double unreduced gametes of the hybrid by normally reduced gametes of one of the parental species.     

Occurrence, inheritance and use of reproductive mutants in alfalfa improvement

The widespread occurrence of 2ngametes (i.e. gametes with the somatic chromosome number) in the Medicago sativa-coerulea-falcata complex supports the concept that gene flow from diploid to tetraploid species occurs continuously in nature and plays a key role in alfalfa evolution. Breeders realized early that gene transfer between ploidy levels via 2n pollen and 2n eggs would have had potential use in cultivated alfalfa improvement. Cytological investigations provided insights into the types of meiotic abnormalities responsible for the production of 2n gametes. Alterations were defined as genetically equivalent to first (FDR) or second division restitution(SDR) mechanisms. For breeding purposes,data have proven that 2n gametes of the FDR type are more advantageous than those obtained by SDR for transferring parental heterozygosity and retaining epistatic interactions. The use of diploid meiotic mutants that produce 2ngametes is now recognized as one of the most effective methods available for exploiting heterosis and introgressing wild germplasm traits into cultivated tetraploid alfalfa via unilateral (USP) and bilateral sexual polyploidization (BSP) schemes. Both2n egg and 2n pollen producers could be used for direct gene transfer from wild diploid relatives into cultivated alfalfa by means of 2x-4x and4x-2x crosses. Although data have shown that forage yield improvement can be achieved when plants are sexually tetraploidized, problems related to reduced plant fertility and seed production remain largely unexplained. Apomixis has the potential of cloning plants through seed and thus provides a unique opportunity for developing superior tetraploid cultivars with permanently fixed heterosis and epistatic effects. A main goal in alfalfa breeding could be the introduction of functional apomixis (i.e. Apomeiosis and parthenogenesis) in cultivated alfalfa stocks. In the future, the efficiency of alfalfa breeding programs based on the use of reproductive mutants could be improved by direct selection at the genotype level using RFLPs and PCR-based markers. Suitable DNA markers and detailed linkage maps of alfalfa mutants should help to discover apomictic mutants and address basic genetic issues such as the extent of genomicre combination in polyploid hybrids and the effect of sexual polyploidization on heterosis. Molecular markers have recently been used in alfalfa for studying the inheritance of 2n gamete formation and identifying polymorphisms associated to genes involved in meiotic abnormalities. Molecular tagging of 2n egg and 2n pollen formation not only should explain the genetic control and regulation of these traits, but may also be an essential step towards marker-assisted selection of 2n gamete producers and implementation of USP and BSP breeding schemes. Future perspectives include strategies for the map-based cloning of genomic DNA markers,and screening of EST mini-libraries related to flowers at different developmental stages from meiotic mutants and wild-type scan lead to the identification of mRNAs and thus of candidate genes that control 2n gamete formation in alfalfa. This revised version was published online in July 2006 with corrections to the Cover Date.     

Phenotypic recurrent selection for 2n pollen and 2n egg production in diploid alfalfa

Corrigendum

Phenotypic recurrent selection for 2n pollen and 2n egg production in diploid alfalfa     

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.     

Effect of selection and temperature stress on the production of 2n gametes in Lotus tenuis

Five Lotus tenuis genotypes showing 2n pollen production greater than 5% and producing 4x progenies in 4x(male sterile Lotus corniculatus) × 2x crosses were found in different natural populations. Meiotic analyses of these plants showed some second metaphases with disoriented spindles, i.e. parallel and tripolar spindles which resulted in diads and triads, respectively. Because useful genes for winter growth and resistance to powdery mildew from L. tenuis can be introgressed into L. corniculatus via 2n gametes, a programme of selection was started to increase the occurrence of 2n gamete production in L. tenuis. Controlled crosses were performed between genotypes producing 2n gametes and the progenies were checked for 2n pollen production and meiotic aberration, following the same methodologies as used for the parental plants. Although in some progeny plants the frequency of 2n gamete production increased remarkably, the overall progeny level of 2n gamete production remained unchanged. This suggested that different genes for meiotic aberration exist in natural populations and/or differential seed-setting ability exists between genotypes. Diallel crosses between selected genotypes confinned this hypothesis. Evaluation of 2n pollen production of clones grown under different temperature regimes (warm vs. temperate) showed that, in L. tenuis, the effects of genotype, temperature and genotype × temperature interaction are significant. In particular, high temperature increases the level of 2n gamete production in L. tenuis.     

Discovery of 2n gametes in tetraploid oat Avena vaviloviana

Summary Sexual polyploidization via the action of 2n gametes (gametes with the sporophytic chromosome number) has been identified as the most important evolutionary mode of polyploidization among plant genera. This study was conducted to determine whether 2n gametes are present in the tetraploid level of the genus Avena (2n=4×=28) Twenty tetraploid Avena lines, representing four species and one interspecific hybrid, were screened for pollen grain size in order to differentiate between n and 2n pollen. Avena vaviloviana (Malz.) Mordv. line PI 412767 was observed to contain large pollen grains at a 1.0% frequency. Cytogenetic analyses of pollen mother cells of PI 412767 revealed cells with double the normal chromosome number (i.e., 56 chromosomes at metaphase I and anaphase I). The mode of chromosome doubling was found to be failure of cell wall formation during the last mitotic division that preceded meiosis. The resulting binucleate cells underwent normal meiotic divisions and formed pollen grains with 28 chromosomes. Based on the formation and function of 2n gametes, three models involving diploid and tetraploid oat lines are proposed to describe possible evolutionary pathways for hexaploid oats. If stable synthetic hexaploid oat lines could be developed by utilizing 2n gametes from diploid and tetraploid oat species through bilateral sexual polyploidization, the resulting hexaploids could be used in breeding programs for transferring genes from diploids and tetraploids to cultivated hexaploids.     

Reciprocal effects in true potato seed breeding in short-day length environments

Reciprocal crosses may have significantly distinct performances regarding tuber characteristics in potato, suggesting the importance of cytoplasmic effects in this crop. The selection of parents for true potato seed breeding therefore needs to consider this potential effect when determining the direction of a cross. The aim of this research was to determine whether a broad‐based true potato seed breeding population, developed at the Centro Internacional de la Papa, could be affected by cytoplasmic effects in the short‐day length environments of the tropics. Two random sets of reciprocal biparental crosses were included in the first set of experiments, which were grown in two contrasting Peruvian locations. Only one out of 14 reciprocal crosses showed significantly distinct performance for tuber yield and tuber set. In the second set, only one of each of the 12 reciprocal crosses had distinct performance for vine earliness, days to flowering and flowering intensity, but four of the reciprocal crosses in the second set showed distinct pollen production. The results suggest that cytoplasmic effects in this breeding population are more important for reproductive characteristics such as pollen production than for tuber yield. This finding is not surprising because male sterility in potato results from the interaction between sensitive cytoplasm and dominant nuclear genes.     

Trifolium ambiguum as a source of variation in the breeding of white clover

Summary Hybrids between T. ambiguum and T. repens were successfully established after ovule culture. The hybrids were male sterile but one of the hybrids produced a single seed when backcrossed to T. repens, while the other produced a significantly greater number of seeds. The majority of the backcross hybrids from the latter had 48 chromosomes, being produced from the fertilisation of unreduced gametes of the F1 hybrid by T. repens pollen. Regular meiosis in the BC1 hybrid resulted in the production of balanced gametes containing the full complement of T. repens chromosomes and the polyhaploid set of T. ambiguum chromosomes. There was no difficulty in establishing a substantial population of BC2 hybrids. The meiotic behaviour of the BC1 hybrid indicated that it should be possible to transfer genes from T. ambiguum into white clover T. repens. The significance of the results in terms of the use of alien variation in the improvement of white clover is discussed.     

果树2n配子研究进展

植物在进化过程中能产生低频率的未减数分裂配子,即2n配子,它在果树育种方面发挥着重要的作用。本文详述了2n配子的主要研究概况、发生途径及影响因素,为其在植物育种、种质利用和种质创新等方面提供了重要的应用基础。同时,通过近年的研究成果,总结出诱导方法和人工分离纯化方法,不同品种可根据其不同的生理生化特性选择合适的方法。另外,也叙述了2n配子在果树方面的特殊应用潜力,即在有性多倍化、稳定传递杂合性、克服自交不亲和和克服胚乳平衡数障碍方面有其他育种方法无法比拟的优势。因此,2n配子育种将会是一种重要的育种途径。     

Pollen viability and meiotic behaviour in intraspecific hybrids of Hydrangea aspera subsp. aspera Kawakami group x subsp. sargentiana

Pollen viability and male meiosis in intraspecific hybrids of Hydrangea aspera subsp. aspera Kawakami group (2n = 2x = 36) and subsp. sargentiana (2n = 2x = 34) were investigated. Although it had been assumed that they were sterile, pollen viability was observed; it varied from 2.5% to 12.1%. The production of gametes with different chromosome numbers was implied by the analysis of the dispersion of the diameter distribution of pollen grains. Analysis of male meiosis made it possible to identify the origins with two major categories of meiotic aberrations: abnormal chromosome distribution (early chromosome migration at metaphase, lagging chromosomes at anaphase, micronuclei at telophase), leading to the formation of unbalanced tetrads and/or ones with supernumerary microspores; and abnormal spindle orientation in metaphase II (tripolar, fused and parallel spindles), leading to the formation of dyads or triads. The mode of 2n pollen formation is of the First Division Restitution type. The high level of parental heterozygosity that is normally associated with them should facilitate the transfer of a polygenic trait in breeding programme.     

Selection on pollen for linolenic acid content in rapeseed, Brassica napus L.

In order to enhance the economic value of edible rapeseed oil, an improvement of quality is necessary. Mutagenesis of rapeseed resulted in a low linolenic acid content and a low ‘linolenic acid (CIS: 3) level to linoleic acid (CIS: 2) level’ ratio, that is, the linoleic desaturation ratio (LDR), in the seeds of the Canadian variety ‘Stellar’. As an early breeding marker for low linolenic acid content, the pollen fatty acid composition was determined on 80 doubled haploid plants derived from a single F₁ hybrid obtained from a cross between ‘Stellar’ and a high CIS: 3 variety ‘Drakkar’. Fatty acid analysis on seed and pollen showed that the low CIS: 3 and the low LDR traits from the ‘Stellar’ variety were expressed in pollen and in seeds, and that a very close correlation (r = 0.88) existed between seed and pollen for these two traits. The inheritance of these traits is controlled by two major genes with additive effects, both in seed and pollen. However, minor genes also appeared to be expressed in pollen and seed. These genes may allow the production of plants with lower CIS: 3 levels than that of the low linolenic acid content parent. The efficiency of this new tool for early screening in breeding programmes is discussed.     

Induction of 2n pollen formation in <Emphasis Type="Italic">Begonia</Emphasis> by trifluralin and N<Subscript>2</Subscript>O treatments

In this study, treatments of both trifluralin (at 10, 100 and 1000 μM) and N₂O (in the form of gas under pressure) were applied to Begonia flower buds to induce the formation of 2n pollen. Three male fertile species (B. cucullata, B. subvillosa var. leptotricha and B. fischeri) and two male sterile hybrids (B. schmidtiana × B. cucullata and B. subvillosa var. leptotricha × B. cucullata) were treated. Pollen size, which is related to pollen DNA content, increased after both N₂O and trifluralin treatments, but the induction of large pollen was genotype dependent. Trifluralin induced large pollen only in the male fertile species, while N₂O treatments induced fertile 2n pollen in the male sterile B. schmidtiana × B. cucullata. Cytological studies showed that trifluralin induced multinuclear monads that resulted in 4n gametes in stead of 2n gametes. In general, large pollen obtained after trifluralin treatments showed low germination capability, while large pollen obtained after N₂O treatments retained high germination capability. Seedlings with raised ploidy level could only be obtained after crosses were performed with large pollen obtained from N₂O treatments. Hence, N₂O treatments are preferable to the use of trifluralin to induce 2n gametes in Begonia.     

普通丝瓜果肉褐变程度的主基因+多基因遗传模型分析

果肉褐变是影响丝瓜品质的重要性状之一,对普通丝瓜果肉褐变程度进行遗传分析,为丝瓜的品质改良奠定基础。以易褐变自交系‘YX014’和耐褐变自交系‘LJ-01’配制成6个世代(P1、P2、F1、F2、B1、B2)为试验材料,应用数量性状主基因+多基因混合遗传模型对丝瓜果肉褐变特性进行遗传模式分析。结果表明：普通丝瓜果肉褐变程度的最佳遗传模型为E-1模型,即由2对加性—显性—上位性主基因+加性—显性多基因控制,且具有耐褐变亲本的优势。B1、B2和F2分离世代的主基因遗传率为81.20%、72.08%和0.52%,多基因遗传率均为0,环境方差占表型方差的比例分别是18.8%、27.92%和99.48%,表明丝瓜果肉褐变程度主要受两对主基因控制,以遗传效应为主,同时受环境影响较大,B1、B2世代主基因选择率高,应在早期世代进行选择。因此,在丝瓜褐变的群体改良中,可对低褐变单株进行定向选择或轮回选择,以提高育种效率。     

Variation in pollen performance among plants of Fagopyrum esculentum

Summary In genetically-heterogeneous outcrossing species, there is the opportunity for selection based on the male gamete. Buckwheat (Fagopyrum esculentum Moench) is self-incompatible with one ovule per flower, so pollen competition at each ovule can be studied. The occurrence of selection among pollen parents was determined, as well as the relative importance of prezygotic and post-zygotic selection. Mixed pollinations from two donors produced nonrandom paternity, with one of the donors being favored about 2:1 on several females. Individual plants showed significant variation in the speed of pollen-tube growth. Therefore, prezygotic selection is likely to have occurred based on the speed of pollen tube growth. In single-donor pollinations, donors had equal success as expected in the absence of post-zygotic selection among donors. However, a significant male x female interaction was found, consistent with postzygotic selection against particular parental combinations. To test whether male fitness is reduced by increased allocation to seed filling during pollen production, large- and small-seeded lines were compared, both as pollen donor and as pollen recipient. The large-seeded line was better in both roles, thus there was no evidence that greater allocation to seeds reduced the quality of the pollen.     

Manipulation of the 2n-gametes frequencies in Solanum pollen

Besides the normal gametes, some 2x potato genotypes produce 2n-gametes as well. In interploidy crosses (2x×4x, 4x×2x) 2n-gametes are necessary, whilst in intercrossing at the diploid level they are undesirable.By means of a set of simply constructed sieves, pollen of several 2n-pollen producing progenitors have been sifted. It was possible to increase the 2n-pollen content of low 2n graded pollen to interesting levels. No decrease in in vitro pollen vitality due to the treatment could be demonstrated. The in vivo pollen vitality of the aqueous pollen suspension was satisfactory to very good under cool weather conditions.     

Genes for pollen fertility restoration in sunflowers

Summary Research on 16 sources of pollen fertility restoration revealed that this character was controlled in eight cases by one single dominant gene, in three cases by two complementary genes, in four cases by three complementary genes and in one case by the cumulative action of two nonallelic dominant genes.The presence of minor genes for restoration in the genotype of the fertile analogue B could effect partial fertility restoration in the cytoplasmic male-sterile line A, especially in late backcrosses, when its genotype becomes saturated with such polygenes.A second major gene Rf ₂ was identified among the investigated monogenic restorer sources.     

Inheritance in seed size of alfalfa: Quantitative analysis and response to selection

Seed size in alfalfa (Medicago saliva L.) has been positively correlated with seedling vigour and early growth, but there were few published reports on inheritance or selection for this trait. The objective of this research was to estimate components of genetic variance for inheritance of alfalfa seed size and determine the most efficient selection method. Components of genetic variance were estimated on seed and pollen plants of ‘BIC-7-WH’ and their progeny arranged in a North Carolina Design II mating design under controlled environmental conditions. Three selection methods, differing in parental control and selection pressure, were used to determine selection response. The seed parent genotype had a major role in determining alfalfa seed size, but the genotype of the seed had no influence. For genetic studies, pollen and seed parent effects on seed size should be measured on seed harvested from progeny plants. Seed size was controlled by additive and non-additive components of genetic variance. Heritability for seed size was 41.3%. Selection for seed size was effective and a significant shift for larger and smaller seed was attained after one cycle of selection.     

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     

The inheritance of a basal branching type in guar

Summary Eight different genotypes of the F₁ hybrids between Oriental × Asiatic lily (Lilium) hybrids (2n = 2x = 24) were treated with nitrous oxide (N₂O) gas under pressure for 24 and 48 hours. At the time of treatment, all plants possessed early meiotic stages in the anthers of the oldest flower buds. The mature flowers from treated plants were monitored for fertility through pollen germination in vitro as well as by using them in crosses with diploid Asiatic hybrids (2n = 2x = 24) both as male and female parents. In five out of the eight genotypes of OA hybrids there was evidence for the production of 2n pollen which germinated in vitro from either one or both treatments. The 2n pollen from three genotypes was successfully used in crosses. In two cases, the treated plants were successfully used as female parents which indicated the formation of 2n (or 2x) egg cells. From an analysis of 41 sexual polyploid progenies obtained from N₂O treated plants it was shown that they were all euploids consisting of 34 triploids (2n = 3x = 36) and seven tetraploids (2n = 4x = 48). A detailed cytological analysis of 12 progeny plants through genomic in situ hybridization (GISH) proved that N₂O had induced first division restitution gametes in most cases and in two cases they produced gametes through indeterminate meiotic restitution. There was evidence for intergenomic recombination in three cases.