Inter-sectional hybrids obtained from reciprocal crosses between Begonia semperflorens (section Begonia) and B. ‘Orange Rubra’ (section Gaerdita × section Pritzelia)

Inter-sectional hybrids were successfully obtained by the reciprocal crosses between 11 cultivars (including 6 diploids and 5 tetraploids) of Begonia semperflorens (SS & SSSS genomes) and B. ‘Orange Rubra’ (RR genome) with the aid of in vitro culture of mature or immature seeds on MS medium containing 0.1 mg l⁻¹ α-naphthylacetic acid, 0.1 mg l⁻¹ 6-benzyladenine, 10 mg l⁻¹ gibberellic acid, 30 g l⁻¹ sucrose and 2.5 g l⁻¹ gellan gum. Embryo rescue as ovary culture with immature seeds 12^th–16^th day after pollination (DAP) generally gave higher efficiency of plantlet formation, but in some cross combinations, culture of mature seeds (30 DAP) resulted in higher yield of plantlets. Flow cytometric analysis revealed that they were consisted of the plants with various genomic combinations (RS, RR, RSS, RRS, RRSS and RRRRSS) as estimated by the DNA contents of both parents. Hybridity of these plants with various genomic combinations including RR was confirmed by random amplified polymorphic DNA analysis. These results suggested that unreduced gamete formation and spontaneous chromosome doubling were involved in the hybrid formation of various ploidy levels and genomic combinations. These hybrids showed various levels of intermediate traits between both parents according to the genomic compositions, and some of them had desirable characters of both parents.     

2n配子在大白菜育种上的应用

对二倍体大白菜的８７个自交系和３个杂交一代材料进行了雌雄２ｎ配子的测定。结果有１２个材料含有２ｎ配子，占总数的１３．３％。其中有５份既含雄性２ｎ配子又含雌性２ｎ配子，占５．６％。用５个不同包球类型并含有２ｎ配子的二倍体大白菜与四倍体大白菜杂交，选育出具有不同熟性，抗逆性强（抗多种病害，抗寒，耐热）、品质优良，口感好，营养成份高，生长速度快，稔性正常，可在生产上应用的四倍体大白菜材料和多育１、２、３     

Evidence of functional unreduced gametes in Trifolium repens L.

The current research reports the first evidence of functional 2n gametes in megaspores of Trifolium repens L. Reciprocal backcrossing of a colchicine doubled hexaploid (2n = 6x = 48) T. repens × T. nigrescens (6x H-6909-5) F₁ hybrid to T. repens produced four seeds from 1700 pollinations. The backcross involving 6x H-6909-5 as the female parent and T. repens as the male resulted in only one seed which was, as expected, a pentaploid (2n = 5x = 40) presumably carrying four genomes of T. repens and one genome of T. nigrescens. On the other hand, those backcrosses which utilised T. repens as the female parent and 6x H-6909-5 as the male parent, resulted in 3 viable seeds. These were unexpectedly all heptaploid (2n = 7x = 56) with presumably six genomes of T. repens and one genome of T. nigrescens. The occurrence of heptaploids from a T. repens × 6x H-6909-5 (4x–6x) cross can only be explained by the union of n (= 3x = 24) pollen from 6x H-6909-5 with 2n (= 4x = 32) eggs from T. repens. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding behavior of yield components and hollow heart in tetraploid-diploid vs. conventionally derived potato hybrids

Summary A factorial mating design with six 4x cultivars as stylar parents mated to three groups of pollen parents (4x cultivars, 4x high protein selections, and 2x S. phureja) was used to study breeding behavior of yield, tuber set, marketable yield and hollow heart in potatoes. Mean yield for the 4x-2x hybrids was equivalent to that of their cultivar parents although yield components, tuber set and average tuber size, were greater and less, respectively, than the cultivar parents. High estimates of general combining ability were found in both pollen and stylar parents for yield, tuber set, marketable yield, number of marketable tubers and hollow heart in all three hybrid populations. Specific combining ability estimates were significant only for yield and hollow heart. Frequency of white-fleshed 4x-2x progeny of five heterozygous yellow-fleshed 2x phureja genotypes was used to confirm cytological identification of a first division restitution mechanism operative in the 2x parents. Although three of these phureja genotypes generated high-yielding progeny, two others produced 4x-2x progeny with mean yield similar to that of hybrids among 4x cultivars.Scientific journal series article 11,601 of the Minnesota Agricultural Experiment Station.     

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.     

The occurrence of 2n-pollen and the origin of sexual polyploids in dihaploid roses (Rosa hybrida L.)

Haploidisation by in situ parthenogenesis of 4x R. hybrida resulted in the production of some dihaploid roses (2n=2x=14) able to produce viable pollen. A cytological study of microsporogenesis revealed that, although the first meiotic reductional division occurred normally, the second (equational) division was characterised by frequent abnormalities which concerned spindle formation and led to unreduced gametes of First Division Restitution (FDR) type. Analysis of the hybrid progeny of a parthenogenetically derived male fertile dihaploid plant revealed a selective advantage of the 2n-pollen, especially in the case of an hybridization with a tetraploid female parent. Moreover, crosses carried out among dihaploid partners always resulted in hybrids with ploidy levels≥ 3x. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Production of 27-, 28-, and 56-chromosome apomictic hybrid derivatives between pearl millet (2n=14) and Pennisetum squamulatum (2n=54)

Summary An interspecific hybridization program designed to transfer gene(s) controlling apomixis from Pennisetum squamulatum Fresen. (2n=6x=54) to induced tetraploid (2n=4x=28) cultivated pearl millet, Pennisetum americanum (L.) Leeke resulted in four offtype plants, two with 27 chromosomes and two with 28 chromosomes. These plants were found among 217 spaced plants established from open-pollinated seed of an apomictic 21-chromosome polyhaploid (2n=21) plant derived from an apomictic interspecific hybrid (2n=41) between tetraploid pearl millet and Pennisetum squamulatum. It appeared that a 21- (or 20-) chromosome unreduced egg from the apomictic polyhaploid united with a 7-chromosome pearl millet (2n=2x=14) gamete to produce a 28- (or 27-) chromosome offspring. Meiotic chromosome behavior was irregular averaging from 3.60 to 4.05 bivalents per microsporocyte in the 27- and 28-chromosome hybrids. The 27- or 28-chromosome hybrids, like the 21-chromosome female parent, shed no pollen, but set from 1.8 to 28 seed per panicle when allowed to outcross with pearl millet. Progeny of the 28-chromosome hybrids were uniform and identical to their respective female parents, indicating that apomixis had been effectively transferred through the egg. In addition, a 56-chromosome plant resulting from chromosome doubling of a 28-chromosome hybrid was identified. Pollen was 68 per cent stainable and the plant averaged 2.3 selfed seeds per panicle. Chromosomes of the 56-chromosome plant paired as bivalents (x=10.67) or associated in multivalents. Three to nine chromosomes remained unpaired at metaphase I. Multiple four-nucleate embryo sacs indicated the 56-chromosome hybrid was an obligate apomict. The production of 27-, 28-, and 56-chromosome hybrid derivatives were the results of interspecific hybridization, haploidization, fertilization of unreduced apomictic eggs, and spontaneous chromosome doubling. These mechanisms resulted in new unique genome combinations between x=7 and x=9 Pennisetum species.     

Breeding for quantitative resistance to potato cyst nematode (Globodera pallida) in tetraploid potatoes using dihaploids and unreduced gametes

Summary Dihaploids were produced from tetraploids resistant to potato cyst nematode (Globodera pallida (Stone)). High levels of resistance were found in the dihaploids and three were used to produce tetraploid progenies by crossing them with susceptible tetraploid cultivars. One dihaploid, PDH505, produced more highly resistant offspring than the other two, PDHs 417 and 418. The latter gave progenies whose levels of resistance were similar to those obtained from susceptible dihaploids crossed with resistant tetraploids.The differences between the progenies of the resistant dihaploids were probably due to different modes of unreduced gamete formation (PDH505 producing gametes by first division restitution (FDR) and PDHs 417 and 418 by second division restitution (SDR)) although cytological studies would be necessary to confirm this. The methods by which dihaploids could be utilised in a tetraploid potato breeding programme are discussed in relation to the mode of unreduced gamete formation.     

Distorted genetic segregation of the transposon mPing at the long arm of chromosome 12 in rice

A class II transposable element, mPing exists in the rice genome ubiquitously and can transpose even in ordinary cultivation conditions. A copy of mPing was identified at the long arm of chromosome 12. In reciprocal backcrossed F₁s between a heterozygote and a homozygote without mPing, the male gametes with this mPing from heterozygotes were transmitted to the next generation at a lower frequency than those without mPing, resulting in distorted genetic segregation in self-fertilized progenies, as well as in F₁s after backcrossing. Pollens with mPing tended to germinate on stigma less than those without mPing. These results, however, could not explain the lower transmission of male gametes with mPing. In addition, no excision of mPing was observed in a homozygote. Thus, it was suggested that male gametes with mPing were eliminated partly from pollination to fertilization by negative competition against male gametes without mPing. Less formation of microspores with mPing in meiosis could also be a cause for the distorted segregation, although this could not be examined. At least two ORFs, whose functions have not been identified, are located near this mPing. It is plausible that either of these ORFs or both are necessary for the normal functioning of male gametes.